Optimal culture of human umbilical cord blood mesenchymal stem cells / 中国实用内科杂志

ABSTRACT

Objective To optimized the culture conditions of the human umbilical cord blood mesenchymal stem cells.Methods The umbilical cord blood was taken in Guangzhou Hua Qiao Hospital between Oct.2004 to Feb.2005.The isolation method,planting density,the time of the first medium changing,the influence of culture medium on the growth of MSC were analyzed. Mesenchymal stem cells were identified using the surface marker by flow cytometry.Osteoblast was identified by Von Kossa staining and alkaline phosphatase staining,adipocyte was identified by Oil Red O staining. Results When the other conditions were the same,the hespan isolation was better than Ficoll isolation;5?10~6/cm~2 was the best planting density;the best first medium changing was on the seventh day at primary culture. Under optimized conditions,the MSC expressed adhesion molecules CD_ 13 ,CD_ 29 and CD_ 44 ,but not antigens of hematopoietic CD_ 34 ,CD_ 45 ,CD_ 14 and not antigens of endothelia CD_ 106 . Exposure of these cells to osteogenic inductive agents resulted in an increase in expression of alkaline phosphatase and the appearance of hydroxyapatite nodules. Incubation with adipogenic inductive agents resulted in morphological change and staining with Oil Red O. Conclusion Mesenchymal stem cells exist in Cord blood,but slower to establish in culture.Cord blood may prove to be a new source of cells for cellular therapeutics.