| 摘要 |
A method of isolating and growing human mesenchymal stem calls (hMSC) by culturing human stem cells in human umbilical cord blood serum. For this purpose blood is collected from the umbilical cord at the time of birth, after the infant is separated from the umbilical cord, and the blood is collected from an umbilical vein free of anticoagulants. The blood is collected in a blood bag having a collecting needle which is inserted into the umbilical vein and the blood is allowed to flow from the vein into the blood bag, and the blood is allowed to clot at room temperature and the bag is transported to a processing area which is a cGMP clean room. The method also includes aspirating Human Mesenchymal Stem Cells (hMSC) present in bone marrow, diluting bone marrow aspirate with tissue culture medium; plating cell suspension into tissue culture flasks so that Human Mesenchymal Stem Cells (hMSC) can adhere for 24 hours; transferring the resulting supernatant to fresh flasks for the remaining Human Mesenchymal Stem Cells (hMSC) to adhere, feeding cultures every 3 days using the medium and cultures reach confluence by 3 weeks, adding 5-aza cytidine reagent to these cultures at final concentration of 5-20 micro molars and allowing the culture to grow for another 3 days.
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