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.