CN108384742A - A kind of novel activated dose and preparation method thereof for immune cell expansion - Google Patents
A kind of novel activated dose and preparation method thereof for immune cell expansion Download PDFInfo
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Abstract
Novel activated dose and preparation method thereof that the invention discloses a kind of for immune cell expansion, described novel activated dose includes following components by weight percent：20 26% biological conductive nanofibers, 1 3% electroctyte, 3 4% gel mixed solutions, 9 11% compound agonists, 12 14% artificial cell films, 0.3 0.8% cell membrane activator, 8 10% specific antibodies, surplus is liquid medium；The present invention loads compound agonist using most biological conductive nanofiber, then artificial cell film is recycled to be wrapped up, after finally utilizing the processing activation of cell membrane activator, binding specificity antibody；Electroctyte and gel mixed solution are planted on another fraction biological conductive nanofiber, then two parts biological conductive nanofiber is placed into constitute immune cell expansion in liquid medium novel activated dose.In short, the in-vitro multiplication rate of immunocyte can be improved 10 15 times by the present invention.
Description
Technical field
The invention belongs to bioengineering and biomedicine field, and in particular to a kind of novel work for immune cell expansion
Agent and preparation method thereof.
Background technology
Immunocyte (immune cell) is being commonly called as leucocyte, including lymphocyte and various phagocytes etc., also special
Refer to the lymphocyte etc. that can be identified antigen, generate specific immune response.Lymphocyte is the basis of immune system, in body
Interior distribution is very extensive, and mainly T lymphocytes, bone-marrow-derived lymphocyte are activated (activation) by antigenic stimulus, and division increases
It grows, specific immune response occurs.In addition to T lymphocytes and bone-marrow-derived lymphocyte, also K lymphocytes and NK lymphocytes, totally four
Type.T lymphocytes are a multi-functional cell masses.In addition to lymphocyte, the cell for participating in immune response also has slurry thin
Born of the same parents, granulocyte, mast cell, antigen presenting cell and mononuclear phagocyte system cell.
However now for the treatment of cancer, the immunocyte of itself can not maintain patient's self immune system
Normal operation, it is therefore desirable to carry out autoimmune cell adoptive therapy, be exactly that patient itself is had into antiviral and antitumor work
Property cell activate in vitro and amplification processing, selectivity generate high-efficiency low-toxicity antiviral and antitumor immunocyte, then,
These cells are fed back into patient's body, to achieve the purpose that remove virus and kill tumour cell.It is large number of in human body
Specific immunocyte of dividing the work plays a significant role in keeping human internal environment.Lymphocyte is the crack portion in immunocyte
Team, and antigen presenting cell is the radar troop in immunocyte.Antigen presenting cell once detects body danger signal, will
Lymphocyte can be passed to, fast reaction and danger zone can be gone to eliminate danger.In tumour and chronic viral liver
Scorching patient's body, immunocyte quantity and vigor are insufficient, cannot detect tumour cell and poisoning intrusion signal in time, canceration
Tumour cell and the virus of invasion cannot be removed in time, and disease is caused further to deteriorate.Cell therapy at this time at activation from
The key of body immunity.I.e. cell therapy offers the suppressed immunocyte of patient's body, mainly lymphocyte and antigen
Cell takes out, and by replication in vitro, activation, so that their quantity and activity is greatly improved, then feeds back to again
Patient's body, to play the effect of antiviral and antitumor.
However the activator effect of immune cell expansion in the prior art is not satisfactory, cultivates the immuning cell number of amplification
Measure low and of poor quality, and the ingredient for the activator used is fed back with immunocyte into patient's body, be easy to cause adverse reaction.
Invention content
For the above technical problem, the present invention provides a kind of novel activated dose for immune cell expansion and its preparation side
Method.
The technical scheme is that：A kind of novel activated dose for immune cell expansion, the novel activated dose of packet
Include following components by weight percent：20-26% biological conductives nanofiber, 1-3% electroctytes, 3-4% gels mixed solution, 9-11%
Compound agonist, 12-14% artificial cells film, 0.3-0.8% cell membranes activator, 8-10% specific antibodies, surplus are training
Support base fluid；Biological conductive nanofiber can be the carrier as compound agonist, additionally it is possible to make immunocyte and electroctyte
It is sought connections with；Electroctyte can Discharge Simulation human body environment's bioelectricity, and immune cell expansion can be stimulated, artificial cell film
Viscosity can be carried out with more specific antibodies after the processing of cell membrane activator, to improve immunocyte and specificity
The Percentage bound of antibody, the final unexpected amplification rate for improving immunocyte.
The compound agonist includes according to weight percent：0.2-0.9% Caerulomycin As, 0.1-0.3% ring spore bacterium
Element, 2-5% specific cysteine proteases, 1.3-2.5% ginkgo flavonoid glycosides, 3.6-5.4% Bilobalides, 0.7-1.4%
Lemna paucicostata, 0.4-0.7% lignans, 3.2-6.7% soluble protein polysaccharide, surplus is interleukin-22；The ultramarine is mould
Plain A has an effective antifungal activity, and Caerulomycin A can significantly increase in T cell the expression of CTLA-4 and then
Lower the expression of CD28；The Bilobalide, specific cysteine protease have immunoregulatory activity, induce immunocyte
Amplification；Ginkgo flavonoid glycoside can make immunocyte be in excitatory state, it can effectively be inhibited to enter suspend mode, and then promote immune thin
Born of the same parents expand；Lemna paucicostata can correct the disorder of immune system；Lignan can immune cell activated receptor active,
And also there is good inoxidizability；Soluble protein is important osmotic adjustment and nutriment, their increase
The water holding capacity that cell can be improved with accumulation, plays a protective role to the living matter and biomembrane of cell, in addition, soluble egg
White polysaccharide can be used as emulsion stabilizer, so that each ingredient in compound agonist is uniformly mixed, and enhance its stability；
The liquid medium includes according to weight percent：4.5-6.8% umbilical cords extract, 1.2-2.6% fibrins
Original, 0.5-1.2% coagulation factors, 0.3-0.6% trilutes, 3.5-5.0%% glutathione, 4.7-5.3%
Chitosan oligosaccharide, 3.1-6.0% taurines, 2.0-4.5% vitamin Cs, 2.2-3.6% zinc citrates, 1.5-3.7% allantoins,
2.6-4.9% unsaturated fatty acids, 7.6-9.4% balanced salt solutions, surplus are the bases DMEM culture medium；Umbilical cord extract energy
It is even to provide nutriment for immunocyte and electroctyte；Allantoin has the function of that antiallergic is releived, and can reduce human body pair
The allergic reaction of immunocyte after amplification in vitro.
Further, the biological conductive nanofiber includes following composition by weight：40-60 parts of carbon nanotubes, 11-13 parts
Poly-dopamine, 7-9 part polypyrrole, 30-50 parts of N-Methyl pyrrolidones, 24-35 parts of fibroin albumens, 13-22 parts of collagens, 8-10 parts
Lactic acid glycolate copolymer, wherein fibroin albumen, collagen, lactic acid glycolate copolymer have good biological ductility, and
It is degradable, harm will not be generated to human body and external cell；Carbon nanotube, poly-dopamine, polypyrrole are conductive material, can
Electroctyte power generation is conducted, and then immunocyte is promoted to be expanded.
Further, the preparation method of the biological conductive nanofiber includes the following steps：
S1：The carbon nanotube, poly-dopamine, polypyrrole are dissolved in the N-Methyl pyrrolidone, formed conductive
Material solution；
S2：It carries out the fibroin albumen, collagen, lactic acid glycolate copolymer to be mixed to get biomaterial solution；By institute
It is uniform using ultrasonic wave progress dispersion mixing with the biomaterial solution to state conductive material solution, it is molten to obtain biological conductive material
Liquid；
S3：Recycle electrostatic spinning technique by the biological conductive material it is molten blow out biological conductive fiber silk；
S4：The biological conductive fiber silk is placed in vacuum drying chamber, low temperature drying 24- at a temperature of 60-80 DEG C
30h is to get biological conductive nanofiber.
Further, any one of the electroctyte in electric eel, electric ray, raad.
Further, the gel mixed solution be by water-setting collagen protein adhesive, hyaluronic acid, 3D Poly L-lactic acid according to
Weight ratio is 3:1:0.5 mixes.
Further, the artificial cell film is made of following components by weight percent：34-40% lecithin, 12-15% courages are solid
Alcohol, four different palmitic acid of 0.1-0.5% ascorbic acid, 1.2-2.3% vitamin-e esters, 2.5-3.6% esters Sodium Pyruvate, 1.3-
1.8% ornithine, 1.0-1.6% proline, 2.8-4.1% distearoylphosphatidylglycerol, surplus are deionized water；The dimension
Raw element E esters have antioxidation；Ornithine, proline can be covered in as hydrophily adjuvant on phospholipid bilayer surface, from
And play the role of protecting liposome.
Further, novel activated dose of the preparation method includes the following steps：
(1) compound agonist is loaded：By the interleukin-22, Caerulomycin A, cyclosporin, specific cysteine proteases
Enzyme, ginkgo flavonoid glycoside, Bilobalide, Lemna paucicostata, lignan, soluble protein polysaccharide are mixed according to aforementioned proportion
It closes, obtains compound agonist；The biological conductive nanofiber is cut into the biological conductive nanofiber stick and profit of 2-5mm again
10-20min is handled with disinfection by ultraviolet light, cancels the biological conductive nanofiber stick of the 70-80% after poison and the compound excitement
Agent is uniformly mixed, and dispersion load is carried out using ultrasonic wave, until compound agonist is supported on biological conductive Nanowire completely
It ties up on stick；
(2) liquid artificial cell film is prepared：Lecithin, cholesterol, four different palmitic acid of ascorbic acid, vitamin-e ester are pressed
It carries out being mixed to form hydrophobic mixture according to aforementioned proportion, the hydrophobic mixture is with 1:The volume ratio of 2-3 is dissolved in mixing
In organic solvent, pre-composition is obtained；The pre-composition is evaporated processing, the mixed organic solvents is removed, obtains phosphatide
Film；The ester Sodium Pyruvate, ornithine, proline, distearoylphosphatidylglycerol are dissolved in the deionized water again, shape
At hydrophilic mixture；The hydrophilic mixture is mixed with the immobilized artificial membrane again, mechanical agitation 3-5min obtains liquid
State artificial cell film；
(3) coating is handled：There is the biological conductive nanofiber stick of compound agonist to be added to the liquid artificial load
In cell membrane, and ultrasound wave packages coating technique is utilized, has the biological conductive nanofiber stick outer layer of compound agonist uniform load
Liquid artificial cell film in cladding obtains coating biological conductive nanofiber stick；
(4) specific antibody is added：The cell membrane activator is added to the coating biological conductive nanofiber stick
In, the specific antibody is added after culture activation 1-2h, specific antibody is made to be carried out with coating biological conductive nanofiber stick
30-60min specific bonds obtain the biological conductive nanofiber stick with antibody, for use；
(5) electroctyte is planted：Detach electroctyte from the electric eel, electric ray, raad, and by the electroctyte with
The gel mixed solution is uniformly mixed according to aforementioned proportion, obtains mixture；And it will be described mixed using cell seeding art
Zoarium is planted on the biological conductive nanofiber stick of remaining 20-30%, cultivates 1-3h, obtains the biology with electroctyte
Conductive-nano-fibers stick, for use；
(6) activator is prepared：By the umbilical cord extract, fibrinogen, coagulation factor, trilute,
Glutathione, chitosan oligosaccharide, taurine, vitamin C, zinc citrate, allantoin, 3D Poly L-lactic acid, unsaturated fatty acid, balance
Salting liquid is added to according to aforementioned proportion in the culture medium of the bases DMEM, forms liquid medium；It again will be described
Biological conductive nanofiber stick with antibody is added separately to described with the biological conductive nanofiber stick with electroctyte
In liquid medium, activator is obtained.
Compared with prior art, beneficial effects of the present invention are：The present invention utilizes most biological conductive nanofiber
Compound agonist is loaded, then artificial cell film is recycled to be wrapped up, the processing of cell membrane activator is finally utilized to live
After change, more specific antibodies are combined on its surface；Electroctyte and gel mixed solution are planted using cell seeding art
On another fraction biological conductive nanofiber, bioelectric stimulation original is formed, then by two parts biological conductive nanofiber
It is placed into novel activated dose that immune cell expansion is constituted in liquid medium.Wherein, the present invention simulates people using electroctyte
Body environmental organism electricity, and immune cell expansion can be stimulated；Also, the Caerulomycin A in compound agonist has effective anti-
Fungi activity, and Caerulomycin A can significantly increase the expression of CTLA-4 and the then expression of downward CD28 in T cell；
Bilobalide, specific cysteine protease have immunoregulatory activity, induce immune cell expansion；Ginkgo flavonoid glycoside can
So that immunocyte is in excitatory state, can effectively inhibit it to enter suspend mode, and then immunocyte is promoted to be expanded；Radix tetrastigme carries
Take object that can correct the disorder of immune system；Lignan can immune cell activated receptor active, and also have good anti-
Oxidisability；Soluble protein is important osmotic adjustment and nutriment, their increase and accumulation can improve cell
Water holding capacity plays a protective role to the living matter and biomembrane of cell, in addition, soluble protein polysaccharide can be used as emulsification surely
Determine agent, so that each ingredient in compound agonist is uniformly mixed, and enhance its stability.In short, the present invention can will exempt from
The in-vitro multiplication rate of epidemic disease cell improves 10-15 times, has positive effect in drug research and clinical treatment.
Specific implementation mode
With reference to specific embodiment, present invention be described in more detail, but embodiment does not limit the scope of the invention,
The scope of the present invention is determined with core content according to claims.
Embodiment 1
A kind of novel activated dose for immune cell expansion, described novel activated dose includes following components by weight percent：20%
Biological conductive nanofiber, 1% electroctyte, 3% gel mixed solution, 9% compound agonist, 12% artificial cell film,
0.3% cell membrane activator, 8% specific antibody, surplus are liquid medium；Wherein, the electroctyte comes from electric eel.Biology
Conductive-nano-fibers can be the carrier as compound agonist, additionally it is possible to immunocyte and electroctyte be made to be sought connections with；Hair
Electric cell can Discharge Simulation human body environment's bioelectricity, and immune cell expansion can be stimulated, artificial cell film is through cell membrane
Viscosity can be carried out with more specific antibodies after activator processing, to improve the combination of immunocyte and specific antibody
Rate, the final unexpected amplification rate for improving immunocyte.
Wherein, the compound agonist includes according to weight percent：0.2% Caerulomycin A, 0.1% cyclosporin,
2% specific cysteine protease, 1.3% ginkgo flavonoid glycoside, 3.6% Bilobalide, 0.7% Lemna paucicostata, 0.4%
Lignan, 3.2% soluble protein polysaccharide, surplus is interleukin-22；The Caerulomycin A has effective antifungal activity, and
And Caerulomycin A can significantly increase the expression of CTLA-4 and the then expression of downward CD28 in T cell；In the gingko
Ester, specific cysteine protease have immunoregulatory activity, induce immune cell expansion；Ginkgo flavonoid glycoside can make to be immunized
Cell is in excitatory state, can effectively inhibit it to enter suspend mode, and then immunocyte is promoted to be expanded；Lemna paucicostata energy
Enough correct the disorder of immune system；Lignan can immune cell activated receptor active, and also have good inoxidizability；
Soluble protein is important osmotic adjustment and nutriment, their increase and accumulation can improve the water conservation energy of cell
Power plays a protective role to the living matter and biomembrane of cell, in addition, soluble protein polysaccharide can be used as emulsion stabilizer,
So that each ingredient in compound agonist is uniformly mixed, and enhances its stability；
Wherein, the liquid medium includes according to weight percent：4.5% umbilical cord extract, 1.2% fibrinogen,
0.5% coagulation factor, 0.3% trilute, 3.5%% glutathione, 4.7% chitosan oligosaccharide, 3.1% taurine,
2.0% vitamin C, 2.2% zinc citrate, 1.5% allantoin, 2.6% unsaturated fatty acid, 7.6% balanced salt solution, surplus
For the bases DMEM culture medium；Umbilical cord extract occasionally can provide nutriment for immunocyte and electroctyte；Allantoin has
There is antiallergic to releive, allergic reaction of the human body to the immunocyte after amplification in vitro can be reduced.
Wherein, the biological conductive nanofiber includes following composition by weight：40 parts of carbon nanotubes, 11 parts of poly-dopamines, 7
Part polypyrrole, 30 parts of N-Methyl pyrrolidones, 24 parts of fibroin albumens, 13 parts of collagens, 8 parts of lactic acid glycolate copolymer, wherein silk
Fibroin, collagen, lactic acid glycolate copolymer have good biological ductility, and degradable, will not be to human body and external
Cell generate harm；Carbon nanotube, poly-dopamine, polypyrrole are conductive material, and electroctyte can generate electricity and conduct,
And then immunocyte is promoted to be expanded.The preparation method of the biological conductive nanofiber includes the following steps：
S1：The carbon nanotube, poly-dopamine, polypyrrole are dissolved in the N-Methyl pyrrolidone, formed conductive
Material solution；
S2：It carries out the fibroin albumen, collagen, lactic acid glycolate copolymer to be mixed to get biomaterial solution；By institute
It is uniform using ultrasonic wave progress dispersion mixing with the biomaterial solution to state conductive material solution, it is molten to obtain biological conductive material
Liquid；
S3：Recycle electrostatic spinning technique by the biological conductive material it is molten blow out biological conductive fiber silk；
S4：The biological conductive fiber silk is placed in vacuum drying chamber, at a temperature of 60 DEG C low temperature drying for 24 hours, i.e.,
Obtain biological conductive nanofiber.
Wherein, the gel mixed solution is by water-setting collagen protein adhesive, hyaluronic acid, 3D Poly L-lactic acid according to weight
Than being 3:1:0.5 mixes.
Wherein, the artificial cell film is made of following components by weight percent：34% lecithin, 12% cholesterol, 0.1% resist
Four different palmitic acid of bad hematic acid, 1.2% vitamin-e ester, 2.5% ester Sodium Pyruvate, 1.3% ornithine, 1.0% proline, 2.8%
Distearoylphosphatidylglycerol, surplus are deionization；Vitamin-e ester described in water has antioxidation；Ornithine, proline are made
It can be covered on phospholipid bilayer surface for hydrophily adjuvant, to play the role of protecting liposome.
A kind of novel activated dose of preparation method for immune cell expansion includes the following steps：
(1) compound agonist is loaded：By the interleukin-22, Caerulomycin A, cyclosporin, specific cysteine proteases
Enzyme, ginkgo flavonoid glycoside, Bilobalide, Lemna paucicostata, lignan, soluble protein polysaccharide are mixed according to aforementioned proportion
It closes, obtains compound agonist；The biological conductive nanofiber is cut into the biological conductive nanofiber stick of 2mm again and is utilized
Disinfection by ultraviolet light handles 10min, cancels 70% biological conductive nanofiber stick after poison and is mixed with the compound agonist
It closes uniformly, dispersion load is carried out using ultrasonic wave, until compound agonist is supported on completely on biological conductive nanofiber stick；
(2) liquid artificial cell film is prepared：Lecithin, cholesterol, four different palmitic acid of ascorbic acid, vitamin-e ester are pressed
It carries out being mixed to form hydrophobic mixture according to aforementioned proportion, the hydrophobic mixture is with 1:2 volume ratio, which is dissolved in, to be mixed with
In solvent, pre-composition is obtained；The pre-composition is evaporated processing, the mixed organic solvents is removed, obtains immobilized artificial membrane；
The ester Sodium Pyruvate, ornithine, proline, distearoylphosphatidylglycerol are dissolved in the deionized water again, formed
Hydrophilic mixture；The hydrophilic mixture is mixed with the immobilized artificial membrane again, mechanical agitation 3min obtains liquid people
Work cell membrane；
(3) coating is handled：There is the biological conductive nanofiber stick of compound agonist to be added to the liquid artificial load
In cell membrane, and ultrasound wave packages coating technique is utilized, has the biological conductive nanofiber stick outer layer of compound agonist uniform load
Liquid artificial cell film in cladding obtains coating biological conductive nanofiber stick；
(4) specific antibody is added：The cell membrane activator is added to the coating biological conductive nanofiber stick
In, the specific antibody is added after culture activation 1h, specific antibody is made to be carried out with coating biological conductive nanofiber stick
30min specific bonds obtain the biological conductive nanofiber stick with antibody, for use；
(5) electroctyte is planted：Electroctyte is detached from the electric eel, and the electroctyte and the gel are mixed
It closes solution to be uniformly mixed according to aforementioned proportion, obtains mixture；And the mixture is planted in using cell seeding art
On remaining 20% biological conductive nanofiber stick, 1h is cultivated, obtains the biological conductive nanofiber stick with electroctyte,
For use；
(6) activator is prepared：By the umbilical cord extract, fibrinogen, coagulation factor, trilute,
Glutathione, chitosan oligosaccharide, taurine, vitamin C, zinc citrate, allantoin, 3D Poly L-lactic acid, unsaturated fatty acid, balance
Salting liquid is added to according to aforementioned proportion in the culture medium of the bases DMEM, forms liquid medium；It again will be described
Biological conductive nanofiber stick with antibody is added separately to described with the biological conductive nanofiber stick with electroctyte
In liquid medium, activator is obtained.
Embodiment 2
A kind of novel activated dose for immune cell expansion, described novel activated dose includes following components by weight percent：23%
Biological conductive nanofiber, 2% electroctyte, 3.5% gel mixed solution, 10% compound agonist, 13% artificial cell film,
0.55% cell membrane activator, 9% specific antibody, surplus are liquid medium；Wherein, the electroctyte comes from electric ray.It is raw
Object conductive-nano-fibers can be the carrier as compound agonist, additionally it is possible to immunocyte and electroctyte be made to be sought connections with；
Electroctyte can Discharge Simulation human body environment's bioelectricity, and immune cell expansion can be stimulated, artificial cell film passes through cell
Viscosity can be carried out with more specific antibodies after the processing of film activator, to improve the knot of immunocyte and specific antibody
Conjunction rate, the final unexpected amplification rate for improving immunocyte.
Wherein, the compound agonist includes according to weight percent：0.22% Caerulomycin A, 0.2% cyclosporin,
3.5% specific cysteine protease, 1.9% ginkgo flavonoid glycoside, 4.5% Bilobalide, 1.0% Lemna paucicostata,
0.55% lignan, 4.9% soluble protein polysaccharide, surplus is interleukin-22；The Caerulomycin A has effective antimycotic
Activity, and Caerulomycin A can significantly increase the expression of CTLA-4 and the then expression of downward CD28 in T cell；It is described
Bilobalide, specific cysteine protease have immunoregulatory activity, induce immune cell expansion；Ginkgo flavonoid glycoside can
So that immunocyte is in excitatory state, can effectively inhibit it to enter suspend mode, and then immunocyte is promoted to be expanded；Radix tetrastigme carries
Take object that can correct the disorder of immune system；Lignan can immune cell activated receptor active, and also have good anti-
Oxidisability；Soluble protein is important osmotic adjustment and nutriment, their increase and accumulation can improve cell
Water holding capacity plays a protective role to the living matter and biomembrane of cell, in addition, soluble protein polysaccharide can be used as emulsification surely
Determine agent, so that each ingredient in compound agonist is uniformly mixed, and enhance its stability；
Wherein, the liquid medium includes according to weight percent：5.65% umbilical cord extract, 1.9% fibrinogen,
0.85% coagulation factor, 0.45% trilute, 4.25%% glutathione, 5.0% chitosan oligosaccharide, 4.55% N of sulphur
Acid, 3.25% vitamin C, 2.9% zinc citrate, 2.6% allantoin, 3.75% unsaturated fatty acid, 8.5% balance salt are molten
Liquid, surplus are the bases DMEM culture medium；Umbilical cord extract occasionally can provide nutriment for immunocyte and electroctyte；Urine
Bursin has the function of that antiallergic is releived, and can reduce allergic reaction of the human body to the immunocyte after amplification in vitro.
Wherein, the biological conductive nanofiber includes following composition by weight：50 parts of carbon nanotubes, 12 parts of poly-dopamines, 8
Part polypyrrole, 40 parts of N-Methyl pyrrolidones, 29.5 parts of fibroin albumens, 17.5 parts of collagens, 9 parts of lactic acid glycolate copolymer,
In, fibroin albumen, collagen, lactic acid glycolate copolymer have good biological ductility, and degradable, will not to human body and
External cell generates harm；Carbon nanotube, poly-dopamine, polypyrrole are conductive material, and electroctyte can generate electricity and carry out
Conduction, and then immunocyte is promoted to be expanded.The preparation method of the biological conductive nanofiber includes the following steps：
S1：The carbon nanotube, poly-dopamine, polypyrrole are dissolved in the N-Methyl pyrrolidone, formed conductive
Material solution；
S2：It carries out the fibroin albumen, collagen, lactic acid glycolate copolymer to be mixed to get biomaterial solution；By institute
It is uniform using ultrasonic wave progress dispersion mixing with the biomaterial solution to state conductive material solution, it is molten to obtain biological conductive material
Liquid；
S3：Recycle electrostatic spinning technique by the biological conductive material it is molten blow out biological conductive fiber silk；
S4：The biological conductive fiber silk is placed in vacuum drying chamber, low temperature drying 27h at a temperature of 70 DEG C, i.e.,
Obtain biological conductive nanofiber.
Wherein, the gel mixed solution is by water-setting collagen protein adhesive, hyaluronic acid, 3D Poly L-lactic acid according to weight
Than being 3:1:0.5 mixes.
Wherein, the artificial cell film is made of following components by weight percent：37% lecithin, 13.5% cholesterol, 0.3%
Four different palmitic acid of ascorbic acid, 1.75% vitamin-e ester, 3.05% ester Sodium Pyruvate, 1.55% ornithine, 1.3% proline,
3.45% distearoylphosphatidylglycerol, surplus are deionization；Vitamin-e ester described in water has antioxidation；Ornithine, dried meat
Propylhomoserin can be covered in as hydrophily adjuvant on phospholipid bilayer surface, to play the role of protecting liposome.
A kind of novel activated dose of preparation method for immune cell expansion includes the following steps：
(1) compound agonist is loaded：By the interleukin-22, Caerulomycin A, cyclosporin, specific cysteine proteases
Enzyme, ginkgo flavonoid glycoside, Bilobalide, Lemna paucicostata, lignan, soluble protein polysaccharide are mixed according to aforementioned proportion
It closes, obtains compound agonist；The biological conductive nanofiber is cut into the biological conductive nanofiber stick and profit of 3.5mm again
15min is handled with disinfection by ultraviolet light, cancels 75% biological conductive nanofiber stick after poison and is carried out with the compound agonist
It is uniformly mixed, dispersion load is carried out using ultrasonic wave, until compound agonist is supported on completely on biological conductive nanofiber stick；
(2) liquid artificial cell film is prepared：Lecithin, cholesterol, four different palmitic acid of ascorbic acid, vitamin-e ester are pressed
It carries out being mixed to form hydrophobic mixture according to aforementioned proportion, the hydrophobic mixture is with 1:2.5 volume ratio is dissolved in mixing
In organic solvent, pre-composition is obtained；The pre-composition is evaporated processing, the mixed organic solvents is removed, obtains phosphatide
Film；The ester Sodium Pyruvate, ornithine, proline, distearoylphosphatidylglycerol are dissolved in the deionized water again, shape
At hydrophilic mixture；The hydrophilic mixture is mixed with the immobilized artificial membrane again, mechanical agitation 4min obtains liquid
Artificial cell film；
(3) coating is handled：There is the biological conductive nanofiber stick of compound agonist to be added to the liquid artificial load
In cell membrane, and ultrasound wave packages coating technique is utilized, has the biological conductive nanofiber stick outer layer of compound agonist uniform load
Liquid artificial cell film in cladding obtains coating biological conductive nanofiber stick；
(4) specific antibody is added：The cell membrane activator is added to the coating biological conductive nanofiber stick
In, the specific antibody is added after culture activation 1.5h, specific antibody is made to be carried out with coating biological conductive nanofiber stick
45min specific bonds obtain the biological conductive nanofiber stick with antibody, for use；
(5) electroctyte is planted：Electroctyte is detached from the electric ray, and the electroctyte and the gel are mixed
It closes solution to be uniformly mixed according to aforementioned proportion, obtains mixture；And the mixture is planted in using cell seeding art
On remaining 25% biological conductive nanofiber stick, 2h is cultivated, obtains the biological conductive nanofiber stick with electroctyte,
For use；
(6) activator is prepared：By the umbilical cord extract, fibrinogen, coagulation factor, trilute,
Glutathione, chitosan oligosaccharide, taurine, vitamin C, zinc citrate, allantoin, 3D Poly L-lactic acid, unsaturated fatty acid, balance
Salting liquid is added to according to aforementioned proportion in the culture medium of the bases DMEM, forms liquid medium；It again will be described
Biological conductive nanofiber stick with antibody is added separately to described with the biological conductive nanofiber stick with electroctyte
In liquid medium, activator is obtained.
Embodiment 3
A kind of novel activated dose for immune cell expansion, described novel activated dose includes following components by weight percent：26%
Biological conductive nanofiber, 3% electroctyte, 4% gel mixed solution, 11% compound agonist, 14% artificial cell film,
0.8% cell membrane activator, 10% specific antibody, surplus are liquid medium；Wherein, the electroctyte comes from raad.It is raw
Object conductive-nano-fibers can be the carrier as compound agonist, additionally it is possible to immunocyte and electroctyte be made to be sought connections with；
Electroctyte can Discharge Simulation human body environment's bioelectricity, and immune cell expansion can be stimulated, artificial cell film passes through cell
Viscosity can be carried out with more specific antibodies after the processing of film activator, to improve the knot of immunocyte and specific antibody
Conjunction rate, the final unexpected amplification rate for improving immunocyte.
Wherein, the compound agonist includes according to weight percent：0.9% Caerulomycin A, 0.3% cyclosporin,
5% specific cysteine protease, 2.5% ginkgo flavonoid glycoside, 5.4% Bilobalide, 1.4% Lemna paucicostata, 0.7%
Lignan, 6.7% soluble protein polysaccharide, surplus is interleukin-22；The Caerulomycin A has effective antifungal activity, and
And Caerulomycin A can significantly increase the expression of CTLA-4 and the then expression of downward CD28 in T cell；In the gingko
Ester, specific cysteine protease have immunoregulatory activity, induce immune cell expansion；Ginkgo flavonoid glycoside can make to be immunized
Cell is in excitatory state, can effectively inhibit it to enter suspend mode, and then immunocyte is promoted to be expanded；Lemna paucicostata energy
Enough correct the disorder of immune system；Lignan can immune cell activated receptor active, and also have good inoxidizability；
Soluble protein is important osmotic adjustment and nutriment, their increase and accumulation can improve the water conservation energy of cell
Power plays a protective role to the living matter and biomembrane of cell, in addition, soluble protein polysaccharide can be used as emulsion stabilizer,
So that each ingredient in compound agonist is uniformly mixed, and enhances its stability；
Wherein, the liquid medium includes according to weight percent：6.8% umbilical cord extract, 2.6% fibrinogen,
1.2% coagulation factor, 0.6% trilute, 5.0%% glutathione, 5.3% chitosan oligosaccharide, 6.0% taurine,
4.5% vitamin C, 3.6% zinc citrate, 3.7% allantoin, 4.9% unsaturated fatty acid, 9.4% balanced salt solution, surplus
For the bases DMEM culture medium；Umbilical cord extract occasionally can provide nutriment for immunocyte and electroctyte；Allantoin has
There is antiallergic to releive, allergic reaction of the human body to the immunocyte after amplification in vitro can be reduced.
Wherein, the biological conductive nanofiber includes following composition by weight：60 parts of carbon nanotubes, 13 parts of poly-dopamines, 9
Part polypyrrole, 50 parts of N-Methyl pyrrolidones, 35 parts of fibroin albumens, 22 parts of collagens, 10 parts of lactic acid glycolate copolymer, wherein
Fibroin albumen, collagen, lactic acid glycolate copolymer have good biological ductility, and degradable, will not be to human body and body
Outer cell generates harm；Carbon nanotube, poly-dopamine, polypyrrole are conductive material, and electroctyte can generate electricity and pass
It leads, and then immunocyte is promoted to be expanded.The preparation method of the biological conductive nanofiber includes the following steps：
S1：The carbon nanotube, poly-dopamine, polypyrrole are dissolved in the N-Methyl pyrrolidone, formed conductive
Material solution；
S2：It carries out the fibroin albumen, collagen, lactic acid glycolate copolymer to be mixed to get biomaterial solution；By institute
It is uniform using ultrasonic wave progress dispersion mixing with the biomaterial solution to state conductive material solution, it is molten to obtain biological conductive material
Liquid；
S3：Recycle electrostatic spinning technique by the biological conductive material it is molten blow out biological conductive fiber silk；
S4：The biological conductive fiber silk is placed in vacuum drying chamber, low temperature drying 30h at a temperature of 80 DEG C, i.e.,
Obtain biological conductive nanofiber.
Wherein, the gel mixed solution is by water-setting collagen protein adhesive, hyaluronic acid, 3D Poly L-lactic acid according to weight
Than being 3:1:0.5 mixes.
Wherein, the artificial cell film is made of following components by weight percent：40% lecithin, 15% cholesterol, 0.5% resist
Four different palmitic acid of bad hematic acid, 2.3% vitamin-e ester, 3.6% ester Sodium Pyruvate, 1.8% ornithine, 1.6% proline, 4.1%
Distearoylphosphatidylglycerol, surplus are deionization；Vitamin-e ester described in water has antioxidation；Ornithine, proline are made
It can be covered on phospholipid bilayer surface for hydrophily adjuvant, to play the role of protecting liposome.
A kind of novel activated dose of preparation method for immune cell expansion includes the following steps：
(1) compound agonist is loaded：By the interleukin-22, Caerulomycin A, cyclosporin, specific cysteine proteases
Enzyme, ginkgo flavonoid glycoside, Bilobalide, Lemna paucicostata, lignan, soluble protein polysaccharide are mixed according to aforementioned proportion
It closes, obtains compound agonist；The biological conductive nanofiber is cut into the biological conductive nanofiber stick of 5mm again and is utilized
Disinfection by ultraviolet light handles 20min, cancels 80% biological conductive nanofiber stick after poison and is mixed with the compound agonist
It closes uniformly, dispersion load is carried out using ultrasonic wave, until compound agonist is supported on completely on biological conductive nanofiber stick；
(2) liquid artificial cell film is prepared：Lecithin, cholesterol, four different palmitic acid of ascorbic acid, vitamin-e ester are pressed
It carries out being mixed to form hydrophobic mixture according to aforementioned proportion, the hydrophobic mixture is with 1:3 volume ratio, which is dissolved in, to be mixed with
In solvent, pre-composition is obtained；The pre-composition is evaporated processing, the mixed organic solvents is removed, obtains immobilized artificial membrane；
The ester Sodium Pyruvate, ornithine, proline, distearoylphosphatidylglycerol are dissolved in the deionized water again, formed
Hydrophilic mixture；The hydrophilic mixture is mixed with the immobilized artificial membrane again, mechanical agitation 5min obtains liquid people
Work cell membrane；
(3) coating is handled：There is the biological conductive nanofiber stick of compound agonist to be added to the liquid artificial load
In cell membrane, and ultrasound wave packages coating technique is utilized, has the biological conductive nanofiber stick outer layer of compound agonist uniform load
Liquid artificial cell film in cladding obtains coating biological conductive nanofiber stick；
(4) specific antibody is added：The cell membrane activator is added to the coating biological conductive nanofiber stick
In, the specific antibody is added after culture activation 2h, specific antibody is made to be carried out with coating biological conductive nanofiber stick
60min specific bonds obtain the biological conductive nanofiber stick with antibody, for use；
(5) electroctyte is planted：Electroctyte is detached from the raad, and the electroctyte and the gel are mixed
It closes solution to be uniformly mixed according to aforementioned proportion, obtains mixture；And the mixture is planted in using cell seeding art
On remaining 30% biological conductive nanofiber stick, 3h is cultivated, obtains the biological conductive nanofiber stick with electroctyte,
For use；
(6) activator is prepared：By the umbilical cord extract, fibrinogen, coagulation factor, trilute,
Glutathione, chitosan oligosaccharide, taurine, vitamin C, zinc citrate, allantoin, 3D Poly L-lactic acid, unsaturated fatty acid, balance
Salting liquid is added to according to aforementioned proportion in the culture medium of the bases DMEM, forms liquid medium；It again will be described
Biological conductive nanofiber stick with antibody is added separately to described with the biological conductive nanofiber stick with electroctyte
In liquid medium, activator is obtained.
Interpretation of result
Using the embodiment of the present invention 1-3 as experimental group 1-3, conventional activating agent is contrast groups, utilizes cell colony technical bid
Accurate and correction of typist's errors formula counts the immunocyte of amplification in vitro, and carries out Statistics Division using SPSS10.0 statistical softwares
Reason, measurement data withIt indicates, comparison among groups variance uses paired t-test, is difference, timing statistics 4 with P ＜ 0.05
Week, statistical result are shown in Table 1.
1 immunocyte amplification in vitro multiple of table (N=5)
As can be seen from the above table, experimental group 1-3 is all remarkably higher than control group in Each point in time, when by the 4th week, Cord blood
Hematopoietic stem cell expansion is up to 10.91-15.31 times, and control group merely adds 3.12 times.
Although the present invention is described and illustrated with reference to its specific embodiment, it will be appreciated by those skilled in the art that
It can be variously modified without departing from the spirit and scope of the present invention, changed and replaced.Therefore, of the invention
It is intended to the scope limitation only by following claims and these claims should be as broadly as possible explained in rational degree.
Claims (7)
1. a kind of novel activated dose for immune cell expansion, which is characterized in that described novel activated dose includes following weight
Component：20-26% biological conductives nanofiber, 1-3% electroctytes, 3-4% gels mixed solution, the compound excitements of 9-11%
Agent, 12-14% artificial cells film, 0.3-0.8% cell membranes activator, 8-10% specific antibodies, surplus is liquid medium；
The compound agonist includes according to weight percent：0.2-0.9% Caerulomycin As, 0.1-0.3% cyclosporins, 2-
5% specific cysteine protease, 1.3-2.5% ginkgo flavonoid glycosides, 3.6-5.4% Bilobalides, 0.7-1.4% radix tetrastigmes
Extract, 0.4-0.7% lignans, 3.2-6.7% soluble protein polysaccharide, surplus is interleukin-22；
The liquid medium includes according to weight percent：4.5-6.8% umbilical cords extract, 1.2-2.6% fibrinogens,
0.5-1.2% coagulation factors, 0.3-0.6% trilutes, 3.5-5.0%% glutathione, 4.7-5.3% shells are few
Sugar, 3.1-6.0% taurines, 2.0-4.5% vitamin Cs, 2.2-3.6% zinc citrates, 1.5-3.7% allantoins, 2.6-
4.9% unsaturated fatty acid, 7.6-9.4% balanced salt solutions, surplus are the bases DMEM culture medium.
2. a kind of novel activated dose for immune cell expansion as described in claim 1, which is characterized in that the biology is led
Conducting nanofiber includes following composition by weight：40-60 parts of carbon nanotubes, 11-13 parts of poly-dopamines, 7-9 parts of polypyrroles, 30-50
Part N-Methyl pyrrolidone, 24-35 parts of fibroin albumens, 13-22 parts of collagens, 8-10 parts of lactic acid glycolate copolymer.
3. a kind of novel activated dose for immune cell expansion as claimed in claim 1 or 2, which is characterized in that the life
The preparation method of object conductive-nano-fibers includes the following steps：
S1：The carbon nanotube, poly-dopamine, polypyrrole are dissolved in the N-Methyl pyrrolidone, conductive material is formed
Solution；
S2：It carries out the fibroin albumen, collagen, lactic acid glycolate copolymer to be mixed to get biomaterial solution；It is led described
Electric material solution is uniform using ultrasonic wave progress dispersion mixing with the biomaterial solution, obtains biological conductive material solution；
S3：Recycle electrostatic spinning technique by the biological conductive material it is molten blow out biological conductive fiber silk；
S4：The biological conductive fiber silk is placed in vacuum drying chamber, low temperature drying 24-30h at a temperature of 60-80 DEG C,
Up to biological conductive nanofiber.
4. a kind of novel activated dose for immune cell expansion as described in claim 1, which is characterized in that the biology is led
Conducting nanofiber includes following composition by weight：40-60 parts of carbon nanotubes, 11-13 parts of poly-dopamines, 7-9 parts of polypyrroles, 30-50
Part N-Methyl pyrrolidone, 24-35 parts of fibroin albumens, 13-22 parts of collagens, 8-10 parts of lactic acid glycolate copolymer.
5. a kind of novel activated dose for immune cell expansion as described in claim 1, which is characterized in that the power generation is carefully
Any one of born of the same parents in electric eel, electric ray, raad.
6. a kind of novel activated dose for immune cell expansion as described in claim 1, which is characterized in that it is described it is artificial carefully
After birth is made of following components by weight percent：34-40% lecithin, 12-15% cholesterol, four different palm fibre of 0.1-0.5% ascorbic acid
Palmitic acid acid, 1.2-2.3% vitamin-e esters, 2.5-3.6% esters Sodium Pyruvate, 1.3-1.8% ornithines, 1.0-1.6% proline,
2.8-4.1% distearoylphosphatidylglycerol, surplus are deionized water.
7. a kind of novel activated dose for immune cell expansion as described in claim 1, which is characterized in that the novel work
The preparation method of agent includes the following steps：
(1) compound agonist is loaded：By the interleukin-22, Caerulomycin A, cyclosporin, specific cysteine protease,
Ginkgo flavonoid glycoside, Bilobalide, Lemna paucicostata, lignan, soluble protein polysaccharide are mixed according to aforementioned proportion, are obtained
To compound agonist；The biological conductive nanofiber is cut into the biological conductive nanofiber stick of 2-5mm again and utilizes purple
10-20min is disinfected in outside line, cancel the biological conductive nanofiber stick of the 70-80% after poison and the compound agonist into
Row is uniformly mixed, and dispersion load is carried out using ultrasonic wave, until compound agonist is supported on biological conductive nanofiber stick completely
On；
(2) liquid artificial cell film is prepared：By lecithin, cholesterol, four different palmitic acid of ascorbic acid, vitamin-e ester according to upper
The ratio of stating carries out being mixed to form hydrophobic mixture, and the hydrophobic mixture is with 1:It is organic that the volume ratio of 2-3 is dissolved in mixing
In solvent, pre-composition is obtained；The pre-composition is evaporated processing, the mixed organic solvents is removed, obtains immobilized artificial membrane；Again
The ester Sodium Pyruvate, ornithine, proline, distearoylphosphatidylglycerol are dissolved in the deionized water, parent is formed
Aqueous mixture；The hydrophilic mixture is mixed with the immobilized artificial membrane again, mechanical agitation 3-5min obtains liquid people
Work cell membrane；
(3) coating is handled：There is the biological conductive nanofiber stick of compound agonist to be added to the liquid artificial cell load
In film, and ultrasound wave packages coating technique is utilized, there is the biological conductive nanofiber stick outer layer of compound agonist uniformly to coat load
Upper liquid artificial cell film obtains coating biological conductive nanofiber stick；
(4) specific antibody is added：The cell membrane activator is added in the coating biological conductive nanofiber stick, is trained
The specific antibody is added after supporting 1-2h, specific antibody is made to carry out 30- with coating biological conductive nanofiber stick
60min specific bonds obtain the biological conductive nanofiber stick with antibody, for use；
(5) electroctyte is planted：Detach electroctyte from the electric eel, electric ray, raad, and by the electroctyte with it is described
Gel mixed solution is uniformly mixed according to aforementioned proportion, obtains mixture；And utilize cell seeding art by the mixture
It is planted on the biological conductive nanofiber stick of remaining 20-30%, cultivates 1-3h, obtain the biological conductive with electroctyte
Nanofiber stick, for use；
(6) activator is prepared：By the umbilical cord extract, fibrinogen, coagulation factor, trilute, paddy Guang
Sweet peptide, chitosan oligosaccharide, taurine, vitamin C, zinc citrate, allantoin, 3D Poly L-lactic acid, unsaturated fatty acid, balance salt are molten
Liquid is added to according to aforementioned proportion in the culture medium of the bases DMEM, forms liquid medium；It is carried again by described
The biological conductive nanofiber stick of antibody is added separately to the culture with the biological conductive nanofiber stick with electroctyte
In base fluid, activator is obtained.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107151646A (en) * | 2017-05-18 | 2017-09-12 | 西安交通大学 | A kind of active bio battery construction method based on electroctyte |
| CN109260509A (en) * | 2018-11-27 | 2019-01-25 | 浙江海洋大学 | A kind of self assembly contact-type anti-bacterial hydrogel dressing and preparation method thereof based on poly-dopamine and chitosan oligosaccharide |
| CN110438074A (en) * | 2019-07-16 | 2019-11-12 | 广州沙艾生物科技有限公司 | A kind of immunocyte recovery culture solution and preparation method thereof |
| CN112326743A (en) * | 2020-11-05 | 2021-02-05 | 重庆医科大学 | C-SF-FA flexible conductive film based on silk fibroin, wearable wound monitoring sensor and preparation method of wearable wound monitoring sensor |
| CN115261321A (en) * | 2022-09-27 | 2022-11-01 | 北京大学口腔医学院 | Method for enhancing anti-tumor function of T lymphocyte and application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104818248A (en) * | 2015-03-25 | 2015-08-05 | 苏州佰通生物科技有限公司 | Immunocyte culture medium, and culture method and application of immunocytes |
-
2018
- 2018-02-11 CN CN201810139917.8A patent/CN108384742A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104818248A (en) * | 2015-03-25 | 2015-08-05 | 苏州佰通生物科技有限公司 | Immunocyte culture medium, and culture method and application of immunocytes |
Non-Patent Citations (9)
| Title |
|---|
| H. ARO,ET AL: "Electrostimulation of Rat Callus Cells and Human Lymphocytes In Vitvo", 《JOURNAL OF ORTHOPEDIC RESEARCH》 * |
| MARIA R LOVE,ET AL: "Effects of electrical stimulation on cell proliferation and apoptosis", 《J CELL PHYSIOL》 * |
| 丁钢强等；: "三叶青对小鼠细胞因子及免疫功能影响研究", 《中国卫生检验杂志 》 * |
| 冯枫等: "电磁刺激对脊髓神经干细胞增殖与分化作用研究", 《中华神经外科疾病研究杂志》 * |
| 唐以军等: "银杏叶制剂对哮喘大鼠T淋巴细胞体外增殖和凋亡的影响", 《中国中西医结合杂志》 * |
| 李武丹等: "聚乳酸/聚苯胺/富勒烯静电纺丝纤维的制备与表征", 《高分子学报》 * |
| 熊一全等: "白细胞介素-2增强淋巴细胞对乳腺癌细胞的杀伤作用", 《中华实验外科杂志》 * |
| 贺文凤等；: "脐血间充质干细胞对淋巴细胞增殖的作用", 《中国组织工程研究与临床康复》 * |
| 赵国义: "氨基酸类饲料添加剂及其应用", 《养殖技术顾问》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107151646A (en) * | 2017-05-18 | 2017-09-12 | 西安交通大学 | A kind of active bio battery construction method based on electroctyte |
| CN107151646B (en) * | 2017-05-18 | 2021-07-09 | 西安交通大学 | Active biological battery construction method based on generating cells |
| CN109260509A (en) * | 2018-11-27 | 2019-01-25 | 浙江海洋大学 | A kind of self assembly contact-type anti-bacterial hydrogel dressing and preparation method thereof based on poly-dopamine and chitosan oligosaccharide |
| CN110438074A (en) * | 2019-07-16 | 2019-11-12 | 广州沙艾生物科技有限公司 | A kind of immunocyte recovery culture solution and preparation method thereof |
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