CN113106063A - Method for in-vitro amplification of NK immune cells - Google Patents
Method for in-vitro amplification of NK immune cells Download PDFInfo
- Publication number
- CN113106063A CN113106063A CN202010117536.7A CN202010117536A CN113106063A CN 113106063 A CN113106063 A CN 113106063A CN 202010117536 A CN202010117536 A CN 202010117536A CN 113106063 A CN113106063 A CN 113106063A
- Authority
- CN
- China
- Prior art keywords
- immune cells
- original
- immune
- cells
- culture medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000002865 immune cell Anatomy 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 26
- 238000000338 in vitro Methods 0.000 title claims abstract description 20
- 230000003321 amplification Effects 0.000 title claims abstract description 13
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 13
- 210000004027 cell Anatomy 0.000 claims abstract description 16
- 210000004369 blood Anatomy 0.000 claims abstract description 14
- 239000008280 blood Substances 0.000 claims abstract description 14
- 239000001963 growth medium Substances 0.000 claims abstract description 14
- 238000012258 culturing Methods 0.000 claims abstract description 13
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 claims abstract description 12
- 239000006143 cell culture medium Substances 0.000 claims abstract description 10
- 230000000694 effects Effects 0.000 claims abstract description 9
- 238000004113 cell culture Methods 0.000 claims abstract description 8
- 239000000411 inducer Substances 0.000 claims abstract description 8
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229960002897 heparin Drugs 0.000 claims abstract description 7
- 229920000669 heparin Polymers 0.000 claims abstract description 7
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 claims abstract description 5
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 claims abstract description 5
- 230000010100 anticoagulation Effects 0.000 claims abstract description 4
- 230000006698 induction Effects 0.000 claims abstract description 4
- 210000000822 natural killer cell Anatomy 0.000 claims description 63
- 230000001939 inductive effect Effects 0.000 claims description 8
- 239000000654 additive Substances 0.000 claims description 7
- 230000004936 stimulating effect Effects 0.000 claims description 6
- 230000010261 cell growth Effects 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 102000008100 Human Serum Albumin Human genes 0.000 claims description 4
- 108091006905 Human Serum Albumin Proteins 0.000 claims description 4
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 claims description 4
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 claims description 4
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 claims description 4
- GOTYRUGSSMKFNF-UHFFFAOYSA-N lenalidomide Chemical compound C1C=2C(N)=CC=CC=2C(=O)N1C1CCC(=O)NC1=O GOTYRUGSSMKFNF-UHFFFAOYSA-N 0.000 claims description 4
- 229960004942 lenalidomide Drugs 0.000 claims description 4
- 230000001737 promoting effect Effects 0.000 claims description 4
- 230000001502 supplementing effect Effects 0.000 claims description 2
- 108090000695 Cytokines Proteins 0.000 abstract description 5
- 102000004127 Cytokines Human genes 0.000 abstract description 5
- 210000001744 T-lymphocyte Anatomy 0.000 abstract description 3
- 230000001976 improved effect Effects 0.000 abstract description 3
- 239000003112 inhibitor Substances 0.000 abstract description 3
- 230000007774 longterm Effects 0.000 abstract description 3
- 230000000813 microbial effect Effects 0.000 abstract description 3
- 102000004169 proteins and genes Human genes 0.000 abstract description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 3
- 230000003833 cell viability Effects 0.000 abstract description 2
- 238000001514 detection method Methods 0.000 abstract description 2
- 230000003313 weakening effect Effects 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract 2
- 238000011177 media preparation Methods 0.000 abstract 1
- 238000002659 cell therapy Methods 0.000 description 4
- 230000007123 defense Effects 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 210000003289 regulatory T cell Anatomy 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000033289 adaptive immune response Effects 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000011134 hematopoietic stem cell transplantation Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 210000004976 peripheral blood cell Anatomy 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000009168 stem cell therapy Methods 0.000 description 1
- 238000011476 stem cell transplantation Methods 0.000 description 1
- 238000009580 stem-cell therapy Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 230000002476 tumorcidal effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0646—Natural killers cells [NK], NKT cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/46—Amines, e.g. putrescine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/105—Insulin-like growth factors [IGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/998—Proteins not provided for elsewhere
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses an NK immune cell in-vitro amplification method, which comprises the steps of heparin anticoagulation conventional blood collection, peripheral blood lymphocyte blood separation, NK immune cell culture medium preparation, original NK immune cell separation, original NK immune cell culture, and induction of original NK immune cells and NK immune cell amplification. The method has the advantages that the culture medium with special components is utilized, the traditional method for slowly culturing the NK immune cells is not needed, the activity of the NK immune cells is improved, the occurrence rate of events such as cell viability weakening and microbial pollution which are possibly caused by long-term in-vitro culture is greatly reduced, the original cultured NK immune cell inducer is PMA + ionomycin, T cells are strongly activated, high-concentration cytokines stay in cells by matching with a protein transport inhibitor, the antibody and flow detection is easy, meanwhile, the culture environment is a breathable cell culture bag, the inside and the outside of the culture bag can be subjected to gas exchange, and the living environment safety of the cells is ensured.
Description
Technical Field
The invention relates to an NK immune cell amplification method, in particular to an NK immune cell in-vitro amplification method, and belongs to the technical field of cell in-vitro amplification.
Background
Natural killer cells (NK) are important immune cells of the body, mainly distributed in peripheral blood, and NK cells, as the first line of defense of the body defense system, can not only exert their main tumoricidal effects in the natural immune system, but also secrete different cytokines and various chemokines to regulate the body's adaptive immune response in the early stage of immune response, are indispensable effector cells for the body to exert immune effects, and thus become cell therapies.
As one of the treatment strategies, cell therapy has been studied for decades, and its branches such as stem cell therapy, immune cell therapy, etc. are becoming more and more widely used in clinical applications. The current cell therapy applied in clinic, such as bone marrow hematopoietic stem cell transplantation, mesenchymal stem cell transplantation and the like, is proved to be a safe and effective treatment means.
The content of NK cells in peripheral blood can not meet clinical requirements, and the NK cells are expanded mainly by stimulation of cytokines at present, but the expanded NK cells prepared by the method have certain difference with clinical requirements. How to provide a culture medium capable of amplifying a large amount of high-activity NK cells is one of the problems to be solved in the field of NK cell in-vitro amplification.
Disclosure of Invention
The present invention has been made to solve the above problems, and an object of the present invention is to provide a method for in vitro expansion of NK immune cells.
The invention realizes the purpose through the following technical scheme: a method for in vitro expansion of NK immune cells comprises the following steps:
step one, performing heparin anticoagulation conventional blood collection, namely adding heparin into a blood collection tube, and performing blood collection;
separating peripheral blood lymphocyte blood, and centrifuging the collected blood to separate peripheral blood lymphocyte;
step three, preparing an NK immune cell culture medium, and mixing a required basic culture medium and additives according to a certain dosage;
step four, separating original NK immune cells, and separating peripheral blood lymphocytes to obtain the original NK immune cells;
step five, culturing the original NK immune cells, and putting the obtained original NK immune cells into a culture medium for culturing;
step six, inducing original NK immune cells, and stimulating and inducing the cultured original NK immune cells to obtain high-activity NK immune cells;
and seventhly, amplifying the NK immune cells, namely placing the obtained NK immune cells in a culture bag, and performing amplification culture on the NK immune cells.
As a still further scheme of the invention: in the third step, the NK immune cell culture medium consists of a basic culture medium containing insulin-like growth factor-I, 8-12g/L of human serum albumin, 2.5-7.5mmol/L of nicotinamide, 0.5-2.0 mu mol/L of lenalidomide and the like.
As a still further scheme of the invention: and in the fifth step, culturing the original NK immune cells in an incubator at 37 ℃ and 5% CO2, and supplementing an NK cell culture medium and other required additives at preset intervals.
As a still further scheme of the invention: and in the sixth step, the cultured original NK immune cells are induced to grow by adopting cell growth promoting agents 3CS-NK-B1 and 3 CS-NK-B2.
As a still further scheme of the invention: and in the sixth step, stimulating the original NK immune cells, adding an inducer to increase the cell density to 10 times of the initial culture concentration, and stopping induction, wherein the inducer is PMA + ionomycin.
As a still further scheme of the invention: and seventhly, when the NK immune cells are amplified, the culture bag containing the NK immune cells is a breathable cell culture bag.
The invention has the beneficial effects that: the method for in vitro amplification of NK immune cells is reasonable in design:
1. the culture medium used for culturing the original NK immune cells consists of additives such as insulin-like growth factor-I, human serum albumin, nicotinamide, lenalidomide and the like, and the activity of the NK immune cells is improved by using the culture medium with special components without the traditional method for slowly culturing the NK immune cells, so that the incidence rate of events such as cell activity weakening, microbial pollution and the like possibly caused by long-term in-vitro culture is greatly reduced;
2. the original NK immune cell inducer after culture is PMA + ionomycin, T cells are strongly activated, high-concentration cytokines are enabled to stay in the cells by being matched with a protein transport inhibitor, detection is easy to be carried out by using antibodies and flow, and meanwhile, the culture environment is a breathable cell culture bag, so that gas exchange can be carried out between the inside and the outside of the cell culture bag, and the living environment safety of the cells is ensured.
Drawings
FIG. 1 is a schematic view of the flow structure of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to FIG. 1, a method for in vitro expansion of NK immune cells comprises the following steps:
step one, performing heparin anticoagulation conventional blood collection, namely adding heparin into a blood collection tube, and performing blood collection;
separating peripheral blood lymphocyte blood, and centrifuging the collected blood to separate peripheral blood lymphocyte;
step three, preparing an NK immune cell culture medium, and mixing a required basic culture medium and additives according to a certain dosage;
step four, separating original NK immune cells, and separating peripheral blood lymphocytes to obtain the original NK immune cells;
step five, culturing the original NK immune cells, and putting the obtained original NK immune cells into a culture medium for culturing;
step six, inducing original NK immune cells, and stimulating and inducing the cultured original NK immune cells to obtain high-activity NK immune cells;
and seventhly, amplifying the NK immune cells, namely placing the obtained NK immune cells in a culture bag, and performing amplification culture on the NK immune cells.
Furthermore, in the third step of the present invention, the NK immune cell culture medium is composed of a basal medium containing insulin-like growth factor-i, 8-12g/L human serum albumin, 2.5-7.5mmol/L nicotinamide, 0.5-2.0 μmol/L lenalidomide, etc., which provides a rapid proliferation environment for cell survival, and the activity of NK immune cells is improved by using a culture medium with special components, thereby greatly reducing the incidence of events such as cell viability reduction, microbial contamination, etc. which may occur due to long-term in vitro culture.
Further, in the fifth step of the present invention, the primary NK immune cells are cultured in an incubator at 37 ℃ and 5% CO2, and NK cell culture medium and other required additives are supplemented at predetermined time intervals, so as to simulate human environment and enable the cells to survive and expand when transplanted into a human body.
Further, in the sixth step of the present invention, the cultured primary NK immune cells are induced to grow by using the cell growth promoting agents 3CS-NK-B1 and 3CS-NK-B2, so as to promote the growth of NK immune cells, and the activity of immune cells is increased by matching with the inducing agent.
Further, in the embodiment of the present invention, in the sixth step, the primary NK immune cells are stimulated, and an inducer is added to increase the cell density to 10 times of the initial culture concentration, so as to stop the induction, wherein the inducer is PMA + ionomycin, so that the cells become regulatory T cells with immunosuppressive function, and the transcription of Foxp3 can be increased, thereby promoting the generation of regulatory T cells, strongly activating the T cells, and coordinating with a protein transport inhibitor, so that high-concentration cytokines stay in the cells, which is easy to detect by using antibodies and flow.
Further, in the seventh embodiment of the present invention, in the NK immune cell expansion, the culture bag containing the NK immune cells is a gas-permeable cell culture bag, so that the cells exchange gas during the culture and expansion.
The working principle is as follows: when the method for in vitro expansion of NK immune cells is used, firstly, peripheral blood lymphocytes are separated from collected blood through centrifugation; separating peripheral blood lymphocytes to obtain original NK immune cells; putting the obtained original NK immune cells into a culture medium for culturing; stimulating and inducing the cultured original NK immune cells, then transferring the cultured original NK immune cells into a breathable cell culture bag, adding an expanding agent for continuous stimulation, culturing until the cell expansion reaches 10 times, collecting the cells, and then selecting to obtain the expanded immune cells.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (6)
1. A method for in vitro amplification of NK immune cells, which is characterized by comprising the following steps: the method comprises the following steps:
step one, performing heparin anticoagulation conventional blood collection, namely adding heparin into a blood collection tube, and performing blood collection;
separating peripheral blood lymphocyte blood, and centrifuging the collected blood to separate peripheral blood lymphocyte;
step three, preparing an NK immune cell culture medium, and mixing a required basic culture medium and additives according to a certain dosage;
step four, separating original NK immune cells, and separating peripheral blood lymphocytes to obtain the original NK immune cells;
step five, culturing the original NK immune cells, and putting the obtained original NK immune cells into a culture medium for culturing;
step six, inducing original NK immune cells, and stimulating and inducing the cultured original NK immune cells to obtain high-activity NK immune cells;
and seventhly, amplifying the NK immune cells, namely placing the obtained NK immune cells in a culture bag, and performing amplification culture on the NK immune cells.
2. The method of claim 1 for in vitro expansion of NK immune cells, wherein the method comprises the following steps: in the third step, the NK immune cell culture medium consists of a basic culture medium containing insulin-like growth factor-I, 8-12g/L of human serum albumin, 2.5-7.5mmol/L of nicotinamide, 0.5-2.0 mu mol/L of lenalidomide and the like.
3. The method of claim 1 for in vitro expansion of NK immune cells, wherein the method comprises the following steps: and in the fifth step, culturing the original NK immune cells in an incubator at 37 ℃ and 5% CO2, and supplementing an NK cell culture medium and other required additives at preset intervals.
4. The method of claim 1 for in vitro expansion of NK immune cells, wherein the method comprises the following steps: and in the sixth step, the cultured original NK immune cells are induced to grow by adopting cell growth promoting agents 3CS-NK-B1 and 3 CS-NK-B2.
5. The method of claim 1 for in vitro expansion of NK immune cells, wherein the method comprises the following steps: and in the sixth step, stimulating the original NK immune cells, adding an inducer to increase the cell density to 10 times of the initial culture concentration, and stopping induction, wherein the inducer is PMA + ionomycin.
6. The method of claim 1 for in vitro expansion of NK immune cells, wherein the method comprises the following steps: and seventhly, when the NK immune cells are amplified, the culture bag containing the NK immune cells is a breathable cell culture bag.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010117536.7A CN113106063A (en) | 2020-02-25 | 2020-02-25 | Method for in-vitro amplification of NK immune cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010117536.7A CN113106063A (en) | 2020-02-25 | 2020-02-25 | Method for in-vitro amplification of NK immune cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113106063A true CN113106063A (en) | 2021-07-13 |
Family
ID=76709182
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010117536.7A Pending CN113106063A (en) | 2020-02-25 | 2020-02-25 | Method for in-vitro amplification of NK immune cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113106063A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114149918A (en) * | 2021-11-29 | 2022-03-08 | 深圳中旭生物科技有限公司 | Methods and compositions for binding NK immune cells |
| CN115505600A (en) * | 2022-09-30 | 2022-12-23 | 北京奇迈永华生物科技有限公司 | Method for efficiently infecting human NK cells by lentiviruses |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754705A (en) * | 2017-02-09 | 2017-05-31 | 广州市鲁诚生物科技有限公司 | A kind of method of NK cell culture mediums and amplification in vitro NK cells |
| CN106754701A (en) * | 2016-12-26 | 2017-05-31 | 弎西艾斯（苏州）生物科技有限公司 | A kind of NK cells in vitro induction, amplification method |
| CN109536455A (en) * | 2018-12-11 | 2019-03-29 | 武汉波睿达生物科技有限公司 | A kind of CAR-NK cell and its preparation method and application |
-
2020
- 2020-02-25 CN CN202010117536.7A patent/CN113106063A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754701A (en) * | 2016-12-26 | 2017-05-31 | 弎西艾斯（苏州）生物科技有限公司 | A kind of NK cells in vitro induction, amplification method |
| CN106754705A (en) * | 2017-02-09 | 2017-05-31 | 广州市鲁诚生物科技有限公司 | A kind of method of NK cell culture mediums and amplification in vitro NK cells |
| CN109536455A (en) * | 2018-12-11 | 2019-03-29 | 武汉波睿达生物科技有限公司 | A kind of CAR-NK cell and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| 倪芳: "IGF-1促进人NK细胞发育及杀伤功能", 《万方》 * |
| 林慧 等: "宫内营养过剩对子代单核细胞及NK细胞免疫反应的影响", 《上海交通大学学报（医学版）》, pages 1 - 1 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114149918A (en) * | 2021-11-29 | 2022-03-08 | 深圳中旭生物科技有限公司 | Methods and compositions for binding NK immune cells |
| CN115505600A (en) * | 2022-09-30 | 2022-12-23 | 北京奇迈永华生物科技有限公司 | Method for efficiently infecting human NK cells by lentiviruses |
| CN115505600B (en) * | 2022-09-30 | 2024-01-30 | 北京奇迈永华生物科技有限公司 | Method for efficiently infecting human NK cells by slow viruses |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Marín Morales et al. | Automated clinical grade expansion of regulatory T cells in a fully closed system | |
| CN103756963A (en) | Method used for in vitro proliferation of NK cells | |
| CN105754942A (en) | Method for amplifying NK cells in vitro and NK cells obtained by same | |
| CN102517253A (en) | In vitro amplification and low-temperature storage method for regulatory T cells of umbilical cord blood | |
| CN105238752A (en) | Culture system and culture method for efficient amplification in vitro of autologous NK cells | |
| CN104938477A (en) | CIK (cytokine-induced killer) frozen stock solution and frozen preservation method | |
| CN113106063A (en) | Method for in-vitro amplification of NK immune cells | |
| EP2859092B1 (en) | Therapeutic vaccine for treatment of diabetes type 1 in children, application of the cell sorter and the method of multiplying treg cells to produce therapeutic vaccine for treatment of diabetes type 1 | |
| JPH10295369A (en) | Production of hematopoietic stem cell | |
| CN112680415A (en) | NK culture medium and amplification culture method thereof | |
| CN114085812B (en) | Mesenchymal stem cell population with high expression of CD106 and/or CD142 and reduced expression, and preparation method and application thereof | |
| CN112662625B (en) | T cell culture medium and method for expanding and culturing T cells by using same | |
| CN109182263A (en) | A method of palace film mescenchymal stem cell is separated with Tea Saponin dissolution menstruation erythrocyte | |
| CN106566807A (en) | Concentration gradient rhIL-2 dependent iNKT cell amplification method and application thereof | |
| CN107090434B (en) | Blood anticoagulation and preservation method for improving CIK cell culture effect | |
| CN110862962A (en) | Method for culturing and amplifying NK cells in vitro by using gallic acid | |
| CN113943704B (en) | Preparation method of tumor neoantigen specific T cells | |
| CN112300992B (en) | NK cell culture solution and multistage activated NK cell culture method | |
| CN116179484A (en) | Preparation method and application of T lymphocyte derived from tumor tissue | |
| CN113481157B (en) | Optimized preparation method of specific antiviral adoptive immune cells | |
| CN111690607B (en) | Efficient killer cell in-vitro culture kit and culture method | |
| CN114807031A (en) | Construction method of human peripheral blood immune cell bank and stem cell bank | |
| CN114058584A (en) | Preparation method of natural killer cells for clinical use | |
| CN107779434A (en) | The experimental method of efficient amplification Human peripheral blood NK cells | |
| CN118773134B (en) | CIK cell culture solution suitable for CIK cell culture and method for culturing CIK cells by using same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210713 |