CN105238754A - 一种高增殖力和高杀伤力nk细胞的体外培养方法 - Google Patents
一种高增殖力和高杀伤力nk细胞的体外培养方法 Download PDFInfo
- Publication number
- CN105238754A CN105238754A CN201510810854.0A CN201510810854A CN105238754A CN 105238754 A CN105238754 A CN 105238754A CN 201510810854 A CN201510810854 A CN 201510810854A CN 105238754 A CN105238754 A CN 105238754A
- Authority
- CN
- China
- Prior art keywords
- cell
- concentration
- human
- amplification
- vitro
- 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.)
- Granted
Links
- 210000000822 natural killer cell Anatomy 0.000 title claims abstract description 52
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000000338 in vitro Methods 0.000 title claims abstract description 20
- 102000003812 Interleukin-15 Human genes 0.000 claims abstract description 20
- 108090000172 Interleukin-15 Proteins 0.000 claims abstract description 20
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 claims abstract description 19
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 claims abstract description 19
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 claims abstract description 19
- 108010002350 Interleukin-2 Proteins 0.000 claims abstract description 17
- 238000004113 cell culture Methods 0.000 claims abstract description 12
- 210000005087 mononuclear cell Anatomy 0.000 claims abstract description 9
- 230000000638 stimulation Effects 0.000 claims abstract description 9
- 210000005259 peripheral blood Anatomy 0.000 claims abstract description 8
- 239000011886 peripheral blood Substances 0.000 claims abstract description 8
- 230000003321 amplification Effects 0.000 claims description 23
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 23
- 239000012679 serum free medium Substances 0.000 claims description 18
- 238000011534 incubation Methods 0.000 claims description 4
- 239000002246 antineoplastic agent Substances 0.000 claims description 2
- 229940041181 antineoplastic drug Drugs 0.000 claims description 2
- 230000004936 stimulating effect Effects 0.000 claims description 2
- 239000011248 coating agent Substances 0.000 abstract description 6
- 238000000576 coating method Methods 0.000 abstract description 6
- 238000009169 immunotherapy Methods 0.000 abstract description 4
- 210000004881 tumor cell Anatomy 0.000 abstract description 3
- 238000012136 culture method Methods 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 34
- 102000000588 Interleukin-2 Human genes 0.000 description 14
- 210000001519 tissue Anatomy 0.000 description 13
- 210000004369 blood Anatomy 0.000 description 12
- 239000008280 blood Substances 0.000 description 12
- 230000000694 effects Effects 0.000 description 11
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 5
- 230000001413 cellular effect Effects 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 102000004127 Cytokines Human genes 0.000 description 4
- 108090000695 Cytokines Proteins 0.000 description 4
- 229920001917 Ficoll Polymers 0.000 description 4
- 102000008100 Human Serum Albumin Human genes 0.000 description 4
- 108091006905 Human Serum Albumin Proteins 0.000 description 4
- 241000237502 Ostreidae Species 0.000 description 4
- 210000000601 blood cell Anatomy 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000013467 fragmentation Methods 0.000 description 4
- 238000006062 fragmentation reaction Methods 0.000 description 4
- 230000002147 killing effect Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 235000020636 oyster Nutrition 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 102000019034 Chemokines Human genes 0.000 description 2
- 108010012236 Chemokines Proteins 0.000 description 2
- 230000033289 adaptive immune response Effects 0.000 description 2
- 230000022534 cell killing Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000012636 effector Substances 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 102000003810 Interleukin-18 Human genes 0.000 description 1
- 108090000171 Interleukin-18 Proteins 0.000 description 1
- 102000000704 Interleukin-7 Human genes 0.000 description 1
- 108010002586 Interleukin-7 Proteins 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000001815 biotherapy Methods 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000000527 lymphocytic effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004719 natural immunity Effects 0.000 description 1
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
培养天数(天) | 0 | 7 | 14 | 21 |
实施例4 | 1 | 12.55±3.26 | 41.72±4.12 | 82.57±4.78 |
实施例1 | 1 | 29.65±3.75** | 79.82±5.17** | 159.95±5.14** |
Claims (10)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510810854.0A CN105238754B (zh) | 2015-11-20 | 2015-11-20 | 一种高增殖力和高杀伤力nk细胞的体外培养方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510810854.0A CN105238754B (zh) | 2015-11-20 | 2015-11-20 | 一种高增殖力和高杀伤力nk细胞的体外培养方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN105238754A true CN105238754A (zh) | 2016-01-13 |
CN105238754B CN105238754B (zh) | 2016-08-17 |
Family
ID=55036574
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201510810854.0A Expired - Fee Related CN105238754B (zh) | 2015-11-20 | 2015-11-20 | 一种高增殖力和高杀伤力nk细胞的体外培养方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN105238754B (zh) |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105907714A (zh) * | 2016-04-28 | 2016-08-31 | 王晓冰 | 一种改良的nk细胞培养方法 |
CN106591233A (zh) * | 2016-12-28 | 2017-04-26 | 广州沙艾生物科技有限公司 | 一种免疫细胞的体外诱导扩增和冻存的方法 |
CN106701681A (zh) * | 2016-12-28 | 2017-05-24 | 广州沙艾生物科技有限公司 | 一种免疫细胞的体外诱导扩增、冻存和复苏的方法 |
CN106834227A (zh) * | 2016-12-26 | 2017-06-13 | 深圳精准医疗科技有限公司 | 自然杀伤细胞的体外纯化方法 |
CN107022524A (zh) * | 2017-03-14 | 2017-08-08 | 上海莱馥生命科学技术有限公司 | 一种从外周血单个核细胞中大量扩增nk细胞的方法 |
CN108220235A (zh) * | 2016-12-22 | 2018-06-29 | 细胞邦(北京)生物技术有限公司 | 一种体外活化扩增人自然杀伤(nk)细胞的方法及其专用培养体系 |
CN108300694A (zh) * | 2018-02-07 | 2018-07-20 | 安徽古生物科技有限公司 | 一种nk细胞的无血清培养方法 |
CN109207424A (zh) * | 2018-10-24 | 2019-01-15 | 河南省肿瘤医院 | 一种免疫细胞的培养方法 |
CN109394785A (zh) * | 2018-09-25 | 2019-03-01 | 天津市康婷生物工程有限公司 | 一种nk细胞预防黑色素瘤发生的实验方法 |
CN111394309A (zh) * | 2019-12-10 | 2020-07-10 | 广东先康达生物科技有限公司 | 一种nk细胞体外扩增培养的方法 |
CN112300992A (zh) * | 2020-10-27 | 2021-02-02 | 杜德(江门)生物科技有限公司 | 一种nk细胞培养液及多级激活nk细胞的培养方法 |
CN113388042A (zh) * | 2021-06-28 | 2021-09-14 | 北京鼎成肽源生物技术有限公司 | 一种重组蛋白、重组表达载体、重组细胞和nk细胞激活磁珠及其制备方法和应用 |
CN114891740A (zh) * | 2022-03-25 | 2022-08-12 | 和携科技有限公司 | 一种高增殖能力和高细胞毒性nk细胞的培养方法及其应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102994449A (zh) * | 2012-12-13 | 2013-03-27 | 上海柯莱逊生物技术有限公司 | 一种体外扩增nk细胞的方法 |
-
2015
- 2015-11-20 CN CN201510810854.0A patent/CN105238754B/zh not_active Expired - Fee Related
Non-Patent Citations (1)
Title |
---|
刘悦明等: "新疆地区健康成人外周血NK细胞体外扩增技术的研究", 《新疆医科大学学报》 * |
Cited By (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105907714A (zh) * | 2016-04-28 | 2016-08-31 | 王晓冰 | 一种改良的nk细胞培养方法 |
CN108220235A (zh) * | 2016-12-22 | 2018-06-29 | 细胞邦(北京)生物技术有限公司 | 一种体外活化扩增人自然杀伤(nk)细胞的方法及其专用培养体系 |
CN106834227A (zh) * | 2016-12-26 | 2017-06-13 | 深圳精准医疗科技有限公司 | 自然杀伤细胞的体外纯化方法 |
CN106591233A (zh) * | 2016-12-28 | 2017-04-26 | 广州沙艾生物科技有限公司 | 一种免疫细胞的体外诱导扩增和冻存的方法 |
CN106701681A (zh) * | 2016-12-28 | 2017-05-24 | 广州沙艾生物科技有限公司 | 一种免疫细胞的体外诱导扩增、冻存和复苏的方法 |
CN107022524B (zh) * | 2017-03-14 | 2019-10-22 | 上海莱馥医疗科技有限公司 | 一种从外周血单个核细胞中大量扩增nk细胞的方法 |
CN107022524A (zh) * | 2017-03-14 | 2017-08-08 | 上海莱馥生命科学技术有限公司 | 一种从外周血单个核细胞中大量扩增nk细胞的方法 |
CN108300694A (zh) * | 2018-02-07 | 2018-07-20 | 安徽古生物科技有限公司 | 一种nk细胞的无血清培养方法 |
CN108300694B (zh) * | 2018-02-07 | 2020-10-20 | 安徽古一生物科技有限公司 | 一种nk细胞的无血清培养方法 |
CN109394785A (zh) * | 2018-09-25 | 2019-03-01 | 天津市康婷生物工程有限公司 | 一种nk细胞预防黑色素瘤发生的实验方法 |
CN109207424A (zh) * | 2018-10-24 | 2019-01-15 | 河南省肿瘤医院 | 一种免疫细胞的培养方法 |
CN111394309A (zh) * | 2019-12-10 | 2020-07-10 | 广东先康达生物科技有限公司 | 一种nk细胞体外扩增培养的方法 |
CN112300992A (zh) * | 2020-10-27 | 2021-02-02 | 杜德(江门)生物科技有限公司 | 一种nk细胞培养液及多级激活nk细胞的培养方法 |
CN113388042A (zh) * | 2021-06-28 | 2021-09-14 | 北京鼎成肽源生物技术有限公司 | 一种重组蛋白、重组表达载体、重组细胞和nk细胞激活磁珠及其制备方法和应用 |
CN113388042B (zh) * | 2021-06-28 | 2022-02-01 | 北京鼎成肽源生物技术有限公司 | 一种重组蛋白、重组表达载体、重组细胞和nk细胞激活磁珠及其制备方法和应用 |
CN114891740A (zh) * | 2022-03-25 | 2022-08-12 | 和携科技有限公司 | 一种高增殖能力和高细胞毒性nk细胞的培养方法及其应用 |
CN114891740B (zh) * | 2022-03-25 | 2023-12-01 | 和携科技有限公司 | 一种高增殖能力和高细胞毒性nk细胞的培养方法及其应用 |
Also Published As
Publication number | Publication date |
---|---|
CN105238754B (zh) | 2016-08-17 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105238754A (zh) | 一种高增殖力和高杀伤力nk细胞的体外培养方法 | |
CN103756963B (zh) | 一种体外扩增nk细胞的方法 | |
CN109294985B (zh) | 一种用于nk细胞体外扩增的培养基体系及nk细胞体外扩增的方法 | |
CN105524880A (zh) | 一种免疫细胞库的构建方法 | |
CN105087488A (zh) | 一种肿瘤抗原诱导的dc-cik细胞的制备方法及应用 | |
CN102676455B (zh) | 脐带血来源的树突状细胞及树突状细胞疫苗的制备方法 | |
CN104593326A (zh) | 一种中药诱导增强型dc-cik细胞的制备方法及其应用 | |
CN104498434A (zh) | 一种大量树突状细胞的制备方法、所得树突状细胞 | |
CN105219713A (zh) | 用于肿瘤过继性免疫的高增殖力、高杀伤力nk细胞 | |
CN104371973B (zh) | 一种免疫细胞的无血清培养基 | |
CN103301449A (zh) | 一种大规模培养树突状细胞疫苗的制备方法及其应用 | |
CN109207424A (zh) | 一种免疫细胞的培养方法 | |
CN104480069A (zh) | 一种利用外周血分离培养免疫细胞的方法 | |
CN104894072A (zh) | 一种自体自然杀伤细胞增殖的制备方法及其应用 | |
CN103981144A (zh) | 自体血清抗原致敏dc-cik细胞的制备方法 | |
CN103013914A (zh) | 体外培养杀伤性t细胞的方法 | |
CN107384860A (zh) | 细胞培养液和nk细胞的培养方法 | |
CN104524560A (zh) | 树突状细胞肿瘤疫苗及其制备方法和用途 | |
CN107502591A (zh) | 一种浓度梯度rhIL‑2依赖的iNKT细胞扩增方法及其应用 | |
CN106047809A (zh) | 一种联合tlr7配体同时扩增人cik/nk细胞的方法 | |
CN105535940A (zh) | 一种治疗多发性骨髓瘤的Vγ9Vδ2T细胞制剂的制备方法 | |
CN105368778A (zh) | 一种联合新型tlr7激动剂gd制备人cik细胞的方法 | |
CN107779435A (zh) | 一种含有自体cik细胞的共培养上清液及其应用 | |
CN105219728A (zh) | 一种用于激活乳腺癌特异性免疫反应的试剂盒 | |
CN106754688A (zh) | 一种冻存外周血单个核细胞的高效复苏方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C41 | Transfer of patent application or patent right or utility model | ||
CB03 | Change of inventor or designer information |
Inventor after: Liu Minglu Inventor after: Wu Dongying Inventor after: Feng Jianhai Inventor after: Zhang Chuanpeng Inventor after: Jin Haifeng Inventor after: Liu Jun Inventor after: Ma Honghua Inventor before: Zhao Shunying |
|
COR | Change of bibliographic data | ||
TA01 | Transfer of patent application right |
Effective date of registration: 20160530 Address after: 261500 Gaomi City high tech Industrial Development Zone, Weifang, Shandong Applicant after: JINAN XINGYI MEDICAL TECHNOLOGY CO., LTD. Address before: 211100 Nanjing Medical University, 818 East Tianyuan Road, Jiangning District, Jiangsu, Nanjing Applicant before: Zhao Shunying |
|
CB03 | Change of inventor or designer information |
Inventor after: Liu Minglu Inventor after: Wu Dongying Inventor after: Li Yanzhi Inventor after: Feng Jianhai Inventor after: Zhang Chuanpeng Inventor after: Jin Haifeng Inventor after: Liu Jun Inventor after: Ma Honghua Inventor before: Liu Minglu Inventor before: Wu Dongying Inventor before: Feng Jianhai Inventor before: Zhang Chuanpeng Inventor before: Jin Haifeng Inventor before: Liu Jun Inventor before: Ma Honghua |
|
COR | Change of bibliographic data | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160817 Termination date: 20171120 |
|
CF01 | Termination of patent right due to non-payment of annual fee |