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Culture method of CD34+ hematopoietic stem cells

A technology of hematopoietic stem cells and culture methods, applied in the biological field, can solve the problems of allogeneic source rejection and infectious diseases, high cost, easy differentiation, etc., and achieve the effect of reducing costs and improving amplification efficiency

Active Publication Date: 2020-01-21
北京默赛尔生物科技有限责任公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for culturing CD34+ hematopoietic stem cells derived from umbilical cord blood, aiming to solve the problems of existing hematopoietic stem cell culturing methods that are prone to differentiation during in vitro expansion, rejection of allogeneic sources, and infectious diseases. and high cost

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  • Culture method of CD34+ hematopoietic stem cells
  • Culture method of CD34+ hematopoietic stem cells
  • Culture method of CD34+ hematopoietic stem cells

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Embodiment 1

[0064] A method for culturing CD34+ hematopoietic stem cells, comprising the following steps:

[0065] Configure complete medium for CD34+ hematopoietic stem cells, the complete medium for CD34+ hematopoietic stem cells includes cytokines and chemical molecules, wherein the cytokines include SCF, Flt-3L, TPO, IL-6, LDL, and the chemical molecules include SR1 and UM171, and the final concentration of each cytokine and chemical molecule: SCF: 50-500ng / ml, Flt-3L: 50-500ng / ml, TPO: 30-300ng / ml, IL-6: 30-300ng / ml, LDL: 5-50ug / ml, SR1: 200-2000nM, UM171: 20-200nM.

[0066] range reference above

[0067] Provide umbilical cord blood, put 25ml of umbilical cord blood into a 50ml centrifuge tube, centrifuge at 3000rpm for 10min, remove the platelet-rich plasma in the upper layer, and transfer to each tube according to the volume ratio of the centrifuged blood sample and PBS solution at a ratio of 1:1 The centrifuge tube was diluted with PBS solution. According to the volume ratio o...

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Abstract

The invention provides a culture method of CD34+ hematopoietic stem cells. The culture method of the CD34+ hematopoietic stem cells comprises the following steps: preparing a CD34+ hematopoietic stemcell complete medium, wherein the CD34+ hematopoietic stem cell complete medium comprises cytokines and chemical molecules, the cytokines comprise SCF, Flt-3L, TPO, IL-6 and LDL, and the chemical molecules comprise SR1 and UM171; providing umbilical cord blood, and separating PBMC from the umbilical cord blood; performing re-suspension on the PBMC by using the CD34+ hematopoietic stem cell complete medium, and carrying out shaking culture; and then, during culture period, daily adding fresh CD34+ hematopoietic stem cell complete medium, and carrying out culturing for 9-12 days.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for culturing CD34+ hematopoietic stem cells. Background technique [0002] Hematopoietic stem cell (HSC) is an adult stem cell in the blood system, which has the ability of continuous self-renewal, and can multidirectionally differentiate and develop into various cell types in the blood system, and can rebuild the hematopoietic and immune functions of various lines for a long time. In clinical treatment, hematopoietic stem cell transplantation is widely used in blood system diseases and autoimmune diseases. In the treatment of other solid tumors, such as lymphoma, germ cell tumor, breast cancer, and small cell lung cancer, it is mainly used in conventional treatment failure or refractory relapse. treatment and patients with poor prognostic factors. The main sources of hematopoietic stem cell transplantation are umbilical cord blood, bone marrow and mobilized peripheral blo...

Claims

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Application Information

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IPC IPC(8): C12N5/0789
CPCC12N5/0647C12N2501/125C12N2501/145C12N2501/2306C12N2501/26C12N2501/998C12N2501/999
Inventor 刘韬李柱周美龄刘兵
Owner 北京默赛尔生物科技有限责任公司
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