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Method for expanding human cord blood hematopoietic stem cells (CB-HSC) by using own umbilical cord mesenchymal stem cells (UCMSC) as stromal cells

A technology of hematopoietic stem cells and mesenchymal stem cells, which is applied in the field of expanding human umbilical cord blood hematopoietic stem cells, can solve the problems of increasing the complexity of transplant donor HLA differences, achieve the effect of increasing the expansion multiple, ensuring cell activity, and avoiding loss

Active Publication Date: 2017-12-08
吉林省太阳鸟再生医学工程有限责任公司
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Problems solved by technology

But bone marrow cells come from other individuals, adding to the complexity of HLA differences among transplant donors

Method used

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  • Method for expanding human cord blood hematopoietic stem cells (CB-HSC) by using own umbilical cord mesenchymal stem cells (UCMSC) as stromal cells
  • Method for expanding human cord blood hematopoietic stem cells (CB-HSC) by using own umbilical cord mesenchymal stem cells (UCMSC) as stromal cells
  • Method for expanding human cord blood hematopoietic stem cells (CB-HSC) by using own umbilical cord mesenchymal stem cells (UCMSC) as stromal cells

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Experimental program
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Embodiment 1

[0042] Example 1 Collection, culture, cryopreservation and recovery of umbilical cord blood mononuclear cells

[0043] 1. Cord Blood Collection

[0044] The umbilical cord blood of healthy and normal newborns is collected, about 40-50ml, and the mononuclear cells are separated within 8-12 hours.

[0045] 2. Isolation of Mononuclear Cells

[0046] Dilute cord blood with 3 times the volume of PBS (containing 0.1% BSA, 0.6% citric acid, pH7.4), spread on Ficoll-Hypaque layering solution, 20ml cell suspension / 15ml Ficoll, and centrifuge at 800g for 30min. The interface layer cells were collected, washed three times with PBS, centrifuged at 500g for 20min for the first time, and centrifuged at 400g for 5min for the second and third times. Resuspended in serum-free medium StemSpan TM SFEMII+StemSpan TM In CD34+ Expansion Supplement, total cord blood cells are obtained. In order to ensure synchronization with the culture of umbilical cord mesenchymal stem cells, the isolated c...

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Abstract

The invention relates to the technical field of cell culture, and in particular to a method for expanding human cord blood hematopoietic stem cells (CB-HSC) by using own umbilical cord mesenchymal stem cells (UCMSC) as stromal cells. The invention provides a method for massively expanding CB-HSCs in vitro, which is characterized in that the own UCMSCs are used as stromal cells to be co-cultured with CB-HSCs, the growth microenvironment of CB-HSCs in vivo is simulated. This method can be used for effectively expanding CB-HSCs, and moreover the expanded cells overcome the defects of easy injury, disintegration and the like of a cytokine amplification method.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for expanding human umbilical cord blood hematopoietic stem cells by using self-umbilical cord mesenchymal stem cells as stromal cells. Background technique [0002] Umbilical cord blood hematopoietic stem cells have the advantages of convenient extraction, high matching tolerance to human leukocyte antigen (HLA), and low incidence of graft-versus-host reactions. Therefore, they may become an ideal alternative source of bone marrow stem cell transplantation. It is considered to be an important resource in the treatment of hematological diseases, especially hematological tumors, in addition to bone marrow transplantation. However, due to the small number of hematopoietic stem cells in a single umbilical cord blood, its wide application in clinical practice is limited. In vitro expansion of umbilical cord blood hematopoietic stem cells (CB-HSCs), increasing the number...

Claims

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Application Information

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IPC IPC(8): C12N5/0789C12N5/0775
CPCC12N5/0647C12N5/0665C12N2500/38C12N2501/10C12N2501/125C12N2501/145C12N2501/2303C12N2501/2306C12N2501/26C12N2502/137
Inventor 张东吉鲍永利李玉新杨晓光易静雯石晓川李首一
Owner 吉林省太阳鸟再生医学工程有限责任公司
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