CN1408850A - Immortal technology for human blood dentritic cell - Google Patents
Immortal technology for human blood dentritic cell Download PDFInfo
- Publication number
- CN1408850A CN1408850A CN01140516A CN01140516A CN1408850A CN 1408850 A CN1408850 A CN 1408850A CN 01140516 A CN01140516 A CN 01140516A CN 01140516 A CN01140516 A CN 01140516A CN 1408850 A CN1408850 A CN 1408850A
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- cell
- technology
- dentritic
- antigen
- immortal
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- 210000004369 blood Anatomy 0.000 title claims abstract description 7
- 239000008280 blood Substances 0.000 title claims abstract description 7
- 210000004027 cell Anatomy 0.000 claims abstract description 14
- 210000003995 blood forming stem cell Anatomy 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims description 8
- 210000004443 dendritic cell Anatomy 0.000 claims description 7
- 230000003321 amplification Effects 0.000 claims description 6
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 6
- 210000005208 blood dendritic cell Anatomy 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 2
- 238000001890 transfection Methods 0.000 claims description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 claims 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 claims 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 claims 1
- 108700042226 ras Genes Proteins 0.000 claims 1
- 230000004936 stimulating effect Effects 0.000 claims 1
- 239000000427 antigen Substances 0.000 abstract description 4
- 210000005259 peripheral blood Anatomy 0.000 abstract description 4
- 239000011886 peripheral blood Substances 0.000 abstract description 4
- 108091007433 antigens Proteins 0.000 abstract description 3
- 102000036639 antigens Human genes 0.000 abstract description 3
- 239000011324 bead Substances 0.000 abstract description 2
- 108700019961 Neoplasm Genes Proteins 0.000 abstract 1
- 102000048850 Neoplasm Genes Human genes 0.000 abstract 1
- 230000008105 immune reaction Effects 0.000 abstract 1
- 238000000338 in vitro Methods 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- 239000003550 marker Substances 0.000 abstract 1
- 238000005728 strengthening Methods 0.000 abstract 1
- 229960005486 vaccine Drugs 0.000 abstract 1
- 102000004388 Interleukin-4 Human genes 0.000 description 3
- 108090000978 Interleukin-4 Proteins 0.000 description 3
- 108700020796 Oncogene Proteins 0.000 description 3
- 210000004700 fetal blood Anatomy 0.000 description 3
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 2
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 2
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 231100000023 Cell-mediated cytotoxicity Toxicity 0.000 description 1
- 206010057250 Cell-mediated cytotoxicity Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000006354 HLA-DR Antigens Human genes 0.000 description 1
- 108010058597 HLA-DR Antigens Proteins 0.000 description 1
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102000018594 Tumour necrosis factor Human genes 0.000 description 1
- 108050007852 Tumour necrosis factor Proteins 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000010370 cell cloning Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000005890 cell-mediated cytotoxicity Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229940028885 interleukin-4 Drugs 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 238000002826 magnetic-activated cell sorting Methods 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Dentritic cell (DC) is special antigen presentating cell capable of strengthening specific immune reaction included and produced by weak antigen and self-antigen. The immortal technology of human blood dentritic cell is based on the technology of separating hemopoietic stem cell and DC from peripheral blood or funic blood. After the hemopoietic stem cell is separated in a monocloning antigen-magentic bead separating system, it is stimulated with some cell factor to clone and differentiate into the dentritic cell and transfected and cultured with C-myc and H-ras cancer gene to immortalize. The immortal dentritic cell is finally identified with marker molecule and cloned. The technology of the present invention makes it possible to in vitro proliferate dentritic cell for industrial production of efficient vaccine.
Description
Two, technical field: biological medicine technology
Three, background technology: dendritic cell isolation technique, hemopoietic stem cell sorting technique
Three, summary of the invention: (dendritic cells is a cell of being responsible for antigen presentation in the biological vivo immuning system DC) to dendritic cell, is one of body critical function cell of immunogen being produced immunne response.DC extensively is present in tissues such as blood, lymph, liver spleen and skin.DC energy mobilizing function lymphocyte induces to produce cell-mediated cytotoxicity, improves the cellular immune level of body.Particularly it can significantly improve poor antigen and autoantigen inductive specific reaction, thereby plays a significant role in antitumor and antiviral immunity.
DC is a cell in latter stage at end, and multiplication capacity is limited, is difficult in external a large amount of amplifications breedings.People's peripheral blood is the main source that separates and cultivate DC with Cord blood.Conventional method is to isolate hemopoietic stem cell or multipotential stem cell from peripheral blood and Cord blood, use various cytokines then, be divided into DC as STEM CELL FACTOR (SCF), granulocyte, macrophage colony stimulating factor (GM-CSF), tumour necrosis factor (TNF-α), interleukin-4 amplifications such as (IL-4) and induced dry-cell.But the obtainable DC limited amount of this method.Need to separate repeatedly, cost is higher, does not satisfy actual demand.The purpose of human blood DC immortalization technology is exactly to transform DC by oncogene, makes the DC cell immortalityization, in external a large amount of easily amplification breedings, satisfies the needs of immunotherapy such as antitumor, antiviral.
Compare with the method for amplification DC with the classification from human blood of routine, present technique adopts oncogene (C-myc, H-ras etc.) to transform DC, makes the DC immortalization, is convenient to amplification and the preservation of DC, has overcome the defective that DC can not increase in a large number.In case obtained this immortalization DC, then do not need isolated dendritic cell from blood repeatedly, reduced cultivation cost, time saving and energy saving, more important is to cultivate with propagation DC and medicinal industryization for mass-producing to lay a good foundation.Four, embodiment: at first use monoclonal antibody-magnetic bead separation system (MACS) from healthy human peripheral blood or Cord blood, to separate and purifying CD34 male hemopoietic stem cell, use the various kinds of cell factor (SCF then, GM-SCF, TNF-α, IL-4 etc.) appropriate combination is induced CD34 cell proliferation and is separated into DC, use oncogene (C-myc simultaneously, H-ras etc.) adopt electroporation method transfection DC and cultivate transfectional cell, adopt limiting dilution assay to make the culturing cell cloning, use significant molecule (the CD1 α of blood DC at last, HLA-DR, B7-1 etc.) identify, and adopt flow cytometry to separate immortalization DC.
Claims (2)
1, a kind of technology that makes people's blood dendritic cell immortalization when being characterized in stimulating amplification to be divided into dendritic cell with ordinary method the hemopoietic stem cell, makes the dendritic cell immortalization with c-myc and H-ras oncogene transfection culturing cell.
2, make the dendritic cell immortalization in non-human blood source with aforesaid method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN01140516A CN1408850A (en) | 2001-09-18 | 2001-09-18 | Immortal technology for human blood dentritic cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN01140516A CN1408850A (en) | 2001-09-18 | 2001-09-18 | Immortal technology for human blood dentritic cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1408850A true CN1408850A (en) | 2003-04-09 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN01140516A Pending CN1408850A (en) | 2001-09-18 | 2001-09-18 | Immortal technology for human blood dentritic cell |
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| Country | Link |
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| CN (1) | CN1408850A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100398641C (en) * | 2003-10-17 | 2008-07-02 | 高斌 | Methods and kits for culturing dendritic cells |
| CN101755045B (en) * | 2007-05-17 | 2014-02-12 | 生物载体株式会社 | Preparation method of dendritic cells |
| CN101330830B (en) * | 2005-10-18 | 2016-01-20 | 国家犹太健康中心 | Conditionally immortalized long-term stem cells and methods of making and using the same |
| CN107043750A (en) * | 2017-06-13 | 2017-08-15 | 北京焕生汇生物技术研究院 | A kind of DC cell inductions kit and DC cell induction cultural method |
| CN108546679A (en) * | 2018-04-23 | 2018-09-18 | 北京翊博普惠生物科技发展有限公司 | The method and its application of large amplification human mature high activity Dendritic Cells in vitro |
| CN110093316A (en) * | 2018-09-30 | 2019-08-06 | 北京鼎成肽源生物技术有限公司 | A kind of construction method of AFF cell |
| CN112601815A (en) * | 2018-08-27 | 2021-04-02 | 迈凯恩技术有限公司 | Method for evaluating anti-infective drugs, vaccines, and the like using immortalized monocytes and induced cells |
| CN112899308A (en) * | 2021-02-05 | 2021-06-04 | 北京鼎成肽源生物技术有限公司 | Method for electrotransfection of immortalized dendritic cells |
-
2001
- 2001-09-18 CN CN01140516A patent/CN1408850A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100398641C (en) * | 2003-10-17 | 2008-07-02 | 高斌 | Methods and kits for culturing dendritic cells |
| CN101330830B (en) * | 2005-10-18 | 2016-01-20 | 国家犹太健康中心 | Conditionally immortalized long-term stem cells and methods of making and using the same |
| CN101755045B (en) * | 2007-05-17 | 2014-02-12 | 生物载体株式会社 | Preparation method of dendritic cells |
| CN107043750A (en) * | 2017-06-13 | 2017-08-15 | 北京焕生汇生物技术研究院 | A kind of DC cell inductions kit and DC cell induction cultural method |
| CN108546679A (en) * | 2018-04-23 | 2018-09-18 | 北京翊博普惠生物科技发展有限公司 | The method and its application of large amplification human mature high activity Dendritic Cells in vitro |
| WO2019205783A1 (en) * | 2018-04-23 | 2019-10-31 | 北京翊博普惠生物科技发展有限公司 | Method for expanding human dc cell, and human dc cell resource library |
| CN112601815A (en) * | 2018-08-27 | 2021-04-02 | 迈凯恩技术有限公司 | Method for evaluating anti-infective drugs, vaccines, and the like using immortalized monocytes and induced cells |
| CN110093316A (en) * | 2018-09-30 | 2019-08-06 | 北京鼎成肽源生物技术有限公司 | A kind of construction method of AFF cell |
| CN112899308A (en) * | 2021-02-05 | 2021-06-04 | 北京鼎成肽源生物技术有限公司 | Method for electrotransfection of immortalized dendritic cells |
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