CN102643782B - 一种人血及骨髓树突状细胞分离、纯化及培养的方法 - Google Patents
一种人血及骨髓树突状细胞分离、纯化及培养的方法 Download PDFInfo
- Publication number
- CN102643782B CN102643782B CN201210114089.5A CN201210114089A CN102643782B CN 102643782 B CN102643782 B CN 102643782B CN 201210114089 A CN201210114089 A CN 201210114089A CN 102643782 B CN102643782 B CN 102643782B
- Authority
- CN
- China
- Prior art keywords
- dendritic cells
- human blood
- bone marrow
- marrow dendritic
- massfraction
- 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
- 210000004443 dendritic cell Anatomy 0.000 title claims abstract description 115
- 210000004369 blood Anatomy 0.000 title claims abstract description 108
- 239000008280 blood Substances 0.000 title claims abstract description 108
- 238000012258 culturing Methods 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 28
- MIKKOBKEXMRYFQ-WZTVWXICSA-N meglumine amidotrizoate Chemical compound C[NH2+]C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO.CC(=O)NC1=C(I)C(NC(C)=O)=C(I)C(C([O-])=O)=C1I MIKKOBKEXMRYFQ-WZTVWXICSA-N 0.000 claims abstract description 28
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims abstract description 24
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims abstract description 24
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims abstract description 12
- 235000017557 sodium bicarbonate Nutrition 0.000 claims abstract description 12
- 229910000029 sodium carbonate Inorganic materials 0.000 claims abstract description 12
- 210000004700 fetal blood Anatomy 0.000 claims abstract description 7
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims abstract description 4
- 102000004877 Insulin Human genes 0.000 claims abstract description 4
- 108090001061 Insulin Proteins 0.000 claims abstract description 4
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims abstract description 4
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims abstract description 4
- 239000002609 medium Substances 0.000 claims abstract description 4
- 235000019154 vitamin C Nutrition 0.000 claims abstract description 4
- 239000011718 vitamin C Substances 0.000 claims abstract description 4
- 210000001185 bone marrow Anatomy 0.000 claims description 100
- 239000007788 liquid Substances 0.000 claims description 49
- 239000000243 solution Substances 0.000 claims description 32
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 239000012153 distilled water Substances 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000012266 salt solution Substances 0.000 claims description 14
- 210000004027 cell Anatomy 0.000 claims description 13
- 235000015097 nutrients Nutrition 0.000 claims description 10
- 239000011521 glass Substances 0.000 claims description 9
- 239000011259 mixed solution Substances 0.000 claims description 9
- 239000000047 product Substances 0.000 claims description 9
- 230000001413 cellular effect Effects 0.000 claims description 7
- 239000004793 Polystyrene Substances 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 6
- 239000011248 coating agent Substances 0.000 claims description 6
- 238000000576 coating method Methods 0.000 claims description 6
- 238000013016 damping Methods 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 6
- 229920002223 polystyrene Polymers 0.000 claims description 6
- POECFFCNUXZPJT-UHFFFAOYSA-M sodium;carbonic acid;hydrogen carbonate Chemical compound [Na+].OC(O)=O.OC([O-])=O POECFFCNUXZPJT-UHFFFAOYSA-M 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 4
- 230000002093 peripheral effect Effects 0.000 claims description 4
- 210000000130 stem cell Anatomy 0.000 claims description 4
- 230000010261 cell growth Effects 0.000 claims description 3
- 239000000835 fiber Substances 0.000 claims description 3
- 239000003102 growth factor Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000011085 pressure filtration Methods 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 239000002002 slurry Substances 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 3
- 238000009736 wetting Methods 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 2
- 210000001519 tissue Anatomy 0.000 claims 1
- 238000004113 cell culture Methods 0.000 abstract description 3
- 238000007796 conventional method Methods 0.000 abstract description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 abstract 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 abstract 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 abstract 1
- 102000018233 Fibroblast Growth Factor Human genes 0.000 abstract 1
- 108050007372 Fibroblast Growth Factor Proteins 0.000 abstract 1
- 229920001612 Hydroxyethyl starch Polymers 0.000 abstract 1
- 229930182816 L-glutamine Natural products 0.000 abstract 1
- 229930182821 L-proline Natural products 0.000 abstract 1
- 229930003268 Vitamin C Natural products 0.000 abstract 1
- 229940126864 fibroblast growth factor Drugs 0.000 abstract 1
- 229940050526 hydroxyethylstarch Drugs 0.000 abstract 1
- 229940125396 insulin Drugs 0.000 abstract 1
- 229960002429 proline Drugs 0.000 abstract 1
- 239000000427 antigen Substances 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 210000003719 b-lymphocyte Anatomy 0.000 description 4
- 241000711408 Murine respirovirus Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 210000000612 antigen-presenting cell Anatomy 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000011194 good manufacturing practice Methods 0.000 description 2
- 210000003563 lymphoid tissue Anatomy 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 108040006856 interleukin-3 receptor activity proteins Proteins 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 210000005210 lymphoid organ Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 239000013618 particulate matter Substances 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims (8)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210114089.5A CN102643782B (zh) | 2012-04-18 | 2012-04-18 | 一种人血及骨髓树突状细胞分离、纯化及培养的方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210114089.5A CN102643782B (zh) | 2012-04-18 | 2012-04-18 | 一种人血及骨髓树突状细胞分离、纯化及培养的方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102643782A CN102643782A (zh) | 2012-08-22 |
CN102643782B true CN102643782B (zh) | 2014-12-24 |
Family
ID=46656859
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201210114089.5A Active CN102643782B (zh) | 2012-04-18 | 2012-04-18 | 一种人血及骨髓树突状细胞分离、纯化及培养的方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102643782B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103667174B (zh) * | 2012-09-07 | 2016-06-22 | 北京东方华辉生物医药科技有限公司 | 一种用于处理生物样本的组合物 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102154201A (zh) * | 2011-01-21 | 2011-08-17 | 唐明淇 | 一种人类骨髓、脐带血或外周血干细胞处理试剂盒及干细胞分离方法 |
CN102250839A (zh) * | 2011-06-16 | 2011-11-23 | 崔慧斐 | 一种通用型细胞处理试剂盒及其应用方法 |
-
2012
- 2012-04-18 CN CN201210114089.5A patent/CN102643782B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN102643782A (zh) | 2012-08-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102268405B (zh) | 自体nk细胞体外活化扩增培养的方法及其专用培养基 | |
CN101045914A (zh) | 体外诱导造血干/祖细胞分化为成熟红细胞的方法与应用 | |
CN113755426A (zh) | 类器官培养体系及类器官的培养方法 | |
CN113699106A (zh) | 一种细胞因子诱导的杀伤细胞的分离和扩增方法 | |
CN113583952B (zh) | 一种提高干细胞外泌体产量的培养液 | |
CN108486039B (zh) | 小分子诱导人脂肪干细胞分化为睾丸间质细胞的方法 | |
CN112342190A (zh) | 一种提高间充质干细胞外泌体产量的方法及其应用 | |
CN116836933B (zh) | 肝胆癌类器官培养液、培养试剂组合及培养方法 | |
CN102643782B (zh) | 一种人血及骨髓树突状细胞分离、纯化及培养的方法 | |
CN110283785A (zh) | 一种γδT-NK细胞共培养的方法 | |
CN107227296B (zh) | 一种骨髓单核细胞分离、鉴定及诱导分化的方法 | |
CN102492654A (zh) | 一种分离人脐带血干细胞的试剂盒及其使用方法 | |
CN108841790A (zh) | 一种胎盘来源的单个核细胞诱导cik细胞的方法 | |
CN107794269A (zh) | 促进基因编辑t细胞活化及扩增的生物膜、制法及应用 | |
CN109957543A (zh) | 利用脐带血大量扩增脐血nk细胞的方法 | |
CN110857435B (zh) | 用于培养从脐血中分离的免疫细胞的培养基及其培养方法 | |
CN108034634B (zh) | 一种从经血中分离宫内膜间充质干细胞的方法 | |
CN112300988A (zh) | 一种基于肿瘤新生抗原的t细胞分选制备技术及其应用 | |
WO2016143808A1 (ja) | スフェロイドの作製方法及び該方法に用いる培地 | |
CN203333673U (zh) | 用于富集、提纯干细胞的试剂盒 | |
CN108690829B (zh) | 一种高效扩增nk细胞的方法 | |
CN104789529A (zh) | 促进小鼠骨髓造血干细胞体外克隆形成及分化能力的方法 | |
CN101041814A (zh) | 一种磁性纳米细胞自动分选装置及其应用 | |
CN110106143B (zh) | Bcl-2小分子抑制剂在制备成熟红细胞中的用途 | |
CN106047807B (zh) | 一种适用于高密度细胞培养体系的免疫细胞的无血清培养基 |
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 | ||
ASS | Succession or assignment of patent right |
Owner name: TIANJIN HANYANG BIOLOGICALS TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: GAO XU Effective date: 20150828 |
|
C41 | Transfer of patent application or patent right or utility model | ||
TR01 | Transfer of patent right |
Effective date of registration: 20150828 Address after: 300192, 236, Causeway Road, 201-216, Tianjin, Nankai District (Science and Technology Park) Patentee after: Tianjin City, Hao Yang biological technology limited liability company Address before: 300111, Tianjin, Nankai District Miyun Road, Wenchuan, building No. 3, No. 4, gate 101 Patentee before: Gao Xu |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20190320 Address after: 300 000 Tianjin Binhai New Area Tianjin Binhai High-tech Zone Huayuan Industrial Zone Patentee after: TIANJIN HAOYANG HUAKE BIOTECHNOLOGY CO., LTD. Address before: 300192 No. 236 Baidi Road, Nankai District, Tianjin 201-216 (Science and Technology Park) Patentee before: Tianjin City, Hao Yang biological technology limited liability company |
|
TR01 | Transfer of patent right |
Effective date of registration: 20210105 Address after: 300 000 Hongqi South Road 268, Nankai District, Tianjin, 502, 503, 513 (Science and Technology Park) Patentee after: TIANJIN HAOYANG BIOLOGICAL PRODUCTS TECHNOLOGY Co.,Ltd. Address before: 300 000 Tianjin Binhai New Area Tianjin Binhai High-tech Zone Huayuan Industrial Zone Patentee before: TIANJIN HAOYANG HUAKE BIOTECHNOLOGY Co.,Ltd. |
|
TR01 | Transfer of patent right |