CN103712967B - 检测nk细胞活性的方法 - Google Patents
检测nk细胞活性的方法 Download PDFInfo
- Publication number
- CN103712967B CN103712967B CN201410005008.7A CN201410005008A CN103712967B CN 103712967 B CN103712967 B CN 103712967B CN 201410005008 A CN201410005008 A CN 201410005008A CN 103712967 B CN103712967 B CN 103712967B
- Authority
- CN
- China
- Prior art keywords
- cell
- target cell
- fluorescent dye
- fluorescent
- target
- 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.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 29
- 230000000694 effects Effects 0.000 title abstract description 20
- 210000004027 cell Anatomy 0.000 claims abstract description 228
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 58
- 210000000822 natural killer cell Anatomy 0.000 claims abstract description 30
- 238000002156 mixing Methods 0.000 claims abstract description 24
- 238000005516 engineering process Methods 0.000 claims abstract description 12
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 12
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 12
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 12
- 238000001890 transfection Methods 0.000 claims abstract description 12
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000000980 acid dye Substances 0.000 claims abstract description 9
- 102000034287 fluorescent proteins Human genes 0.000 claims abstract description 5
- 108091006047 fluorescent proteins Proteins 0.000 claims abstract description 5
- MURGITYSBWUQTI-UHFFFAOYSA-N fluorescin Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC=C(O)C=C2OC2=CC(O)=CC=C21 MURGITYSBWUQTI-UHFFFAOYSA-N 0.000 claims description 52
- 230000006907 apoptotic process Effects 0.000 claims description 48
- 238000001514 detection method Methods 0.000 claims description 31
- 206010027336 Menstruation delayed Diseases 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 22
- 108010043121 Green Fluorescent Proteins Proteins 0.000 claims description 16
- 238000004043 dyeing Methods 0.000 claims description 15
- 102000004144 Green Fluorescent Proteins Human genes 0.000 claims description 14
- 239000005090 green fluorescent protein Substances 0.000 claims description 14
- 241000700605 Viruses Species 0.000 claims description 12
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 claims description 9
- 108090000623 proteins and genes Proteins 0.000 claims description 8
- DPKHZNPWBDQZCN-UHFFFAOYSA-N acridine orange free base Chemical compound C1=CC(N(C)C)=CC2=NC3=CC(N(C)C)=CC=C3C=C21 DPKHZNPWBDQZCN-UHFFFAOYSA-N 0.000 claims description 4
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 claims description 4
- 108010082025 cyan fluorescent protein Proteins 0.000 claims description 4
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 claims description 4
- 229960005542 ethidium bromide Drugs 0.000 claims description 4
- 238000012856 packing Methods 0.000 claims description 4
- 150000003904 phospholipids Chemical class 0.000 claims description 4
- 239000013600 plasmid vector Substances 0.000 claims description 4
- 108010054624 red fluorescent protein Proteins 0.000 claims description 4
- YXHLJMWYDTXDHS-IRFLANFNSA-N 7-aminoactinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=C(N)C=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 YXHLJMWYDTXDHS-IRFLANFNSA-N 0.000 claims description 3
- 108700012813 7-aminoactinomycin D Proteins 0.000 claims description 3
- 108010004729 Phycoerythrin Proteins 0.000 claims description 3
- 108010004469 allophycocyanin Proteins 0.000 claims description 3
- 108091005948 blue fluorescent proteins Proteins 0.000 claims description 3
- 238000010276 construction Methods 0.000 claims description 3
- 208000035250 cutaneous malignant susceptibility to 1 melanoma Diseases 0.000 claims description 3
- 201000001441 melanoma Diseases 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 230000001640 apoptogenic effect Effects 0.000 abstract description 8
- 102000036213 phospholipid binding proteins Human genes 0.000 abstract 1
- 108091011000 phospholipid binding proteins Proteins 0.000 abstract 1
- 210000000170 cell membrane Anatomy 0.000 description 23
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 10
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 10
- 210000004369 blood Anatomy 0.000 description 8
- 239000008280 blood Substances 0.000 description 8
- 239000012636 effector Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 238000000684 flow cytometry Methods 0.000 description 6
- 102000000412 Annexin Human genes 0.000 description 5
- 108050008874 Annexin Proteins 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000003550 marker Substances 0.000 description 5
- 102000004121 Annexin A5 Human genes 0.000 description 4
- 108090000672 Annexin A5 Proteins 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000012148 binding buffer Substances 0.000 description 3
- 230000022534 cell killing Effects 0.000 description 3
- 238000004737 colorimetric analysis Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000012447 hatching Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 239000003822 epoxy resin Substances 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 229920000647 polyepoxide Polymers 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000009870 specific binding Effects 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000003146 transient transfection Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- QGZCUOLOTMJILH-UHFFFAOYSA-N 2h-tetrazol-2-ium;bromide Chemical compound [Br-].C1=N[NH+]=NN1 QGZCUOLOTMJILH-UHFFFAOYSA-N 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 239000012099 Alexa Fluor family Substances 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010023126 Jaundice Diseases 0.000 description 1
- 241000713666 Lentivirus Species 0.000 description 1
- 241000736199 Paeonia Species 0.000 description 1
- 235000006484 Paeonia officinalis Nutrition 0.000 description 1
- 238000011529 RT qPCR Methods 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
- 208000036142 Viral infection Diseases 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010598 annexinV-PE /7AAD assay Methods 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000012921 fluorescence analysis Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000009610 hypersensitivity Effects 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000003153 stable transfection Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims (3)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410005008.7A CN103712967B (zh) | 2013-12-27 | 2014-01-06 | 检测nk细胞活性的方法 |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310741395.6 | 2013-12-27 | ||
CN201310741395 | 2013-12-27 | ||
CN201410005008.7A CN103712967B (zh) | 2013-12-27 | 2014-01-06 | 检测nk细胞活性的方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN103712967A CN103712967A (zh) | 2014-04-09 |
CN103712967B true CN103712967B (zh) | 2014-12-03 |
Family
ID=50406113
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410005008.7A Active CN103712967B (zh) | 2013-12-27 | 2014-01-06 | 检测nk细胞活性的方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN103712967B (zh) |
Families Citing this family (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101926166B1 (ko) * | 2016-05-24 | 2018-12-06 | 재단법인 아산사회복지재단 | 수용체 시너지 활성을 이용한 nk 세포의 활성도 검사 방법 및 이를 이용한 nk 세포의 활성도가 관련된 질환의 진단 방법 |
CN108333152B (zh) * | 2017-01-19 | 2020-12-22 | 中国科学院烟台海岸带研究所 | 基于基因重组藻蓝蛋白mac和氧化石墨烯量子点的生物素化抗体传感器及其制备方法 |
CN107367455B (zh) * | 2017-07-10 | 2020-10-16 | 中国科学院合肥物质科学研究院 | 一种肿瘤细胞耐受促凋亡蛋白质分子数目绝对定量的测量方法 |
CN111073947A (zh) * | 2018-10-19 | 2020-04-28 | 三峡大学 | 细胞凋亡检测试剂盒及其检测方法和应用 |
CN110018143B (zh) * | 2019-03-29 | 2022-04-05 | 深圳大学 | 检测甲藻细胞凋亡的方法 |
CN110068529B (zh) * | 2019-05-05 | 2023-06-23 | 中国科学院深圳先进技术研究院 | 一种外囊泡的表征方法 |
CN110231275B (zh) * | 2019-05-20 | 2021-05-28 | 西安交通大学 | 一种基于流式细胞术检测nk细胞杀伤活性的方法 |
CN110542675B (zh) * | 2019-09-30 | 2021-12-14 | 广州市锐博生物科技有限公司 | 细胞凋亡检测方法及细胞凋亡检测试剂盒 |
CN112813133B (zh) * | 2021-01-29 | 2022-07-15 | 上海睿钰生物科技有限公司 | 一种细胞杀伤效力的检测方法、***及其应用 |
CN113155712A (zh) * | 2021-05-17 | 2021-07-23 | 上海轩锋生物科技有限公司 | 一种通过流式细胞仪检测细胞活性的方法 |
US20230176039A1 (en) * | 2021-06-01 | 2023-06-08 | Huimin Zhang | A staining method for live-cell imaging |
CN114774467B (zh) * | 2022-04-08 | 2024-03-29 | 华中科技大学 | 一种多重荧光标记活细胞样品的制备方法 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080254480A1 (en) * | 2007-04-11 | 2008-10-16 | University Of Pittsburgh - Of The Commonwealth System Of Higher Education | Microcytoxicity assay by pre-labeling target cells |
MX336473B (es) * | 2010-02-11 | 2016-01-20 | Hoffmann La Roche | Ensayo mediante adcc nk facs tridimensional. |
-
2014
- 2014-01-06 CN CN201410005008.7A patent/CN103712967B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN103712967A (zh) | 2014-04-09 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN103712967B (zh) | 检测nk细胞活性的方法 | |
Adan et al. | Flow cytometry: basic principles and applications | |
Shapiro | Microbial analysis at the single-cell level: tasks and techniques | |
Caron et al. | Assessment of bacterial viability status by flow cytometry and single cell sorting | |
Chan et al. | Rapid image-based cytometry for comparison of fluorescent viability staining methods | |
Yang et al. | Development of an ultrasensitive dual-channel flow cytometer for the individual analysis of nanosized particles and biomolecules | |
Palková et al. | Single‐cell analysis of yeast, mammalian cells, and fungal spores with a microfluidic pressure‐driven chip‐based system | |
CN104198357B (zh) | 流式细胞术方法在疫苗生产实时监测中的应用 | |
US20100221769A1 (en) | Electroporative flow cytometry | |
CN110231275A (zh) | 一种基于流式细胞术检测nk细胞杀伤活性的方法 | |
CN110018143B (zh) | 检测甲藻细胞凋亡的方法 | |
Balke et al. | Visualizing oxidative cellular stress induced by nanoparticles in the subcytotoxic range using fluorescence lifetime imaging | |
CN102947266A (zh) | 分析细胞或含有核酸的其它生物材料的方法 | |
Lanigan et al. | Real time visualization of cancer cell death, survival and proliferation using fluorochrome-transfected cells in an IncuCyte® imaging system | |
Dashkova et al. | Microalgal cytometric analysis in the presence of endogenous autofluorescent pigments | |
CA2321203A1 (en) | Selective cell analysis | |
El‐Hajjar et al. | A guide to flow cytometry: components, basic principles, experimental design, and cancer research applications | |
FI92076B (fi) | Lymfosyyttien tunnusmerkillinen potentiaalille herkkien aineiden sitominen | |
CN103189495A (zh) | 用于检测和分离物质的多相流动*** | |
Kwok et al. | Laser particle barcoding for multi-pass high-dimensional flow cytometry | |
Bantly et al. | CellVue® Claret, a new far-red dye, facilitates polychromatic assessment of immune cell proliferation | |
CN115266664B (zh) | 一种免疫细胞治疗制剂体外杀伤效力评价方法及其应用 | |
CN104515758B (zh) | 促癌活性的定量检测方法以及促癌剂的筛选方法 | |
Lekshmi et al. | A real‐time image‐based approach to distinguish and discriminate apoptosis from necrosis | |
CN107299128A (zh) | 一种检测细胞内aldh活性的试剂盒及方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20181105 Address after: 101109 Beijing Chaoyang District Beijing Shun Tung Street 6 hospital 26 Building 1 stories 101, 2 layer 201, 3 layer 301, 4 layer 401. Patentee after: BEIJING JINYU MEDICAL EXAMINATION LABORATORY Co.,Ltd. Address before: 100050 Yongan Road, Xuanwu District, Beijing 95 Patentee before: BEIJING FRIENDSHIP HOSPITAL, CAPITAL MEDICAL University |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20191128 Address after: 510000 room 706, No. 10, helix 3 Road, international bio Island, Huangpu District, Guangzhou, Guangdong Province Patentee after: Guangzhou jinbilai Biotechnology Co.,Ltd. Address before: 101109 Beijing Chaoyang District Beijing Shun Tung Street 6 hospital 26 Building 1 stories 101, 2 layer 201, 3 layer 301, 4 layer 401. Patentee before: BEIJING JINYU MEDICAL EXAMINATION LABORATORY Co.,Ltd. |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20200311 Address after: Room 406, room 101, floor 1, building 55, yard 5, Yujing East Road, Tongzhou District, Beijing 100000 Patentee after: BEIJING BEIKEWEI BIOLOTY TECHNOLOGY Co.,Ltd. Address before: 510000 room 706, No. 10, helix 3 Road, international bio Island, Huangpu District, Guangzhou, Guangdong Province Patentee before: Guangzhou jinbilai Biotechnology Co.,Ltd. |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20200929 Address after: 100000 No. 95, Yongan Road, Beijing, Xicheng District Patentee after: Wang Zhao Address before: Room 406, room 101, floor 1, building 55, yard 5, Yujing East Road, Tongzhou District, Beijing 100000 Patentee before: BEIJING BEIKEWEI BIOLOTY TECHNOLOGY Co.,Ltd. |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20231114 Address after: 301700 room 106, building G6, Changyuan road international enterprise community, Wuqing Development Zone, Wuqing District, Tianjin Patentee after: Becoway (Tianjin) Biotechnology Co.,Ltd. Address before: 100000 Yongan Road, Xicheng District, Beijing 95 Patentee before: Wang Zhao |