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
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[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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