Isolation,culture and characterization of mesenchymal stem cells derived from Wharton's jelly of human umbilical cord / 解放军医学杂志

ABSTRACT

Objective To investigate the bionomics of mesenchymal stem cells (MSCs) derived from Wharton's jelly of human umbilical cord. Methods Umbilical arteries, vein and umbilical cord tunica externa were removed, and the remaining tissue (Wharton's jelly) was harvested. To gather MSCs, the umbilical cord was cut into small fragments and digested with 0.075% type Ⅰ collagenase, or the small fragments were cultured with DMEM. The cells derived from Wharton's jelly of umbilical cord were serially subcultivated, the growth curve was drawn, and the identification of living cells was made for studying the cell growth kinetics. The surface antigens were detected with flow cytometry, the cartilage markers were detected by histochemistry and immunohistochemistry, and the expressions of Sox-9 and Col-2A1 mRNA were analyzed by RT-PCR. Results The confluence time of primary culture cells was 3-5d in cells harvested with enzyme digestion, and 10-15d in cells harvested with micro mass. The results of flow cytometry showed that the MSCs derived from umbilical cord expressed CD44 and CD105; the expression of HLA-ABC was positive, while HLA-DPDQDR was negative. There was no significant change on the immunophenotype of umbilical cord MSCs after cryopreservation and thawing. The findings of histochemistry and immunohistochemistry showed that the expression of chondrocyte markers in Wharton's jelly MSCs was weakly positive. The results of RT-PCR showed the chondrocyte markers Sox-9 and Col-2A1 were positively expressed in Wharton's jelly MSCs. Conclusion The MSCs derived from Wharton jelly of human umbilical cord do not differentiate into hematopoietic cells, may express the chondrocyte markers (they are suggested to have characteristics of pre-chondrocytes), and is expected to be a new type of stem cells of tissue engineering cartilage.