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Proliferation method for hematopoietic stem cells of umbilical cord blood

A technology for hematopoietic stem cells and umbilical cord blood, applied in the field of cells, can solve the problems of bottlenecks in the formation ability and no breakthrough support, and achieve the effects of simple operation, increasing the total number of cells, and excellent CFU colony formation ability.

Inactive Publication Date: 2018-01-26
广州市天河诺亚生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the expansion system of pure cytokine combination has encountered a bottleneck in both the final number of HSCs and the ability of CFU-GM colony formation, which has no breakthrough support for the wider use of UCB-HSCs in the future

Method used

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  • Proliferation method for hematopoietic stem cells of umbilical cord blood
  • Proliferation method for hematopoietic stem cells of umbilical cord blood
  • Proliferation method for hematopoietic stem cells of umbilical cord blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Stemspan medium was purchased from STEMCELL company; cell protection agent was pure DMSO solution; umbilical cord blood was collected from healthy pregnant women and infants, and tested for hepatitis B, hepatitis C, syphilis, AIDS, cytomegalovirus, TORCH detection, mycoplasma, chlamydia , G-6PD and thalassemia were all negative. The transport conditions of the specimens from collection to transport back to the Guangdong Provincial Cord Blood Bank were kept at 4-8°C. Proceed with hematopoietic stem cell expansion as follows:

[0041] 1. Preparation of QS11 solution and cell expansion medium

[0042] QS11 (≥99%, GC), dissolved in the cell protection agent, prepared into a concentration of 10mmol / L QS11 storage solution;

[0043] Cell expansion medium: Add cytokine combination in Stemspan medium: IL-6 (concentration in cell expansion medium is 20ng / mL), IL-3 (concentration in cell expansion medium is 20ng / mL) mL), TPO (50 ng / mL in cell expansion medium), SCF (50 ng / mL i...

Embodiment 2

[0055] The specific experimental details of Example 2 are the same as those of Example 1 above, except that in the cell expansion medium containing QS11, the concentration of QS11 in the cell expansion medium is 0.5 μmol / L.

Embodiment 3

[0057] The specific experimental details of Example 3 are the same as those of Example 1 above, except that in the cell expansion medium containing QS11, the concentration of QS11 in the cell expansion medium is 1 μmol / L.

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Abstract

The invention belongs to the technical field of cells, and specifically relates to a proliferation method of hematopoietic stem cells. According to the method, the hematopoietic stem cells of umbilical cord blood are used as sample cells, and a cell proliferation culture medium containing QS11 is used for proliferation culture, so as to obtain the hematopoietic stem cells. Due to use of the cell proliferation culture medium, proliferation culture of the hematopoietic stem cells is more efficient, easier and safer. In proliferation culture of the hematopoietic stem cells of umbilical cord blood, both cell factors and QS11 are added, so that more hematopoietic stem cells are generated, and the formation capability of CFU colonies is enhanced. The proliferation method is easy to operate, is low-cost, can produce more hematopoietic stem cells, is safer, and has a stronger capability of building the blood system cells.

Description

technical field [0001] The invention belongs to the field of cell technology, and in particular relates to a method for expanding umbilical cord blood hematopoietic stem cells. Background technique [0002] In the 1950s, Thomas et al. discovered that bone marrow-derived hematopoietic stem cells (Hematopoietic stem cells, HSCs) could rebuild the hematopoietic system, thereby curing a variety of malignant blood diseases including leukemia and multiple myeloma (MDS), and then HSCs were clinically used worldwide. Treatment is widely carried out. With the development of medicine, the research in the field of cell biology and molecular biology continues to deepen. The clinical indications of HSC are constantly being discovered, and the medical value of HSC is becoming more and more prominent. [0003] At present, there are three sources of HSC. In 1956, Thomas performed the first HSC transplantation using adult bone marrow (Bonemarrow, BM)-derived HSC; in 1989, Gluckman and Kurtzb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789
Inventor 嵐山芮魏伟袁嘉恩
Owner 广州市天河诺亚生物工程有限公司