CN1408850A - Immortal technology for human blood dentritic cell - Google Patents
Immortal technology for human blood dentritic cell Download PDFInfo
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- 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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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 |
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| CN1408850A true CN1408850A (en) | 2003-04-09 |
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| CN01140516A Pending CN1408850A (en) | 2001-09-18 | 2001-09-18 | Immortal technology for human blood dentritic cell |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100398641C (en) * | 2003-10-17 | 2008-07-02 | 高斌 | Dendritic cell culturing method and kit |
| CN101755045B (en) * | 2007-05-17 | 2014-02-12 | 生物载体株式会社 | Method for production of dendritic cell |
| CN101330830B (en) * | 2005-10-18 | 2016-01-20 | 国家犹太健康中心 | Condition immortalization long-term stem cells and preparation and use the method for described cell |
| 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 | Dendritic cell culturing method and kit |
| CN101330830B (en) * | 2005-10-18 | 2016-01-20 | 国家犹太健康中心 | Condition immortalization long-term stem cells and preparation and use the method for described cell |
| CN101755045B (en) * | 2007-05-17 | 2014-02-12 | 生物载体株式会社 | Method for production of dendritic cell |
| 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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