ABSTRACT
BACKGROUND:Human placenta is a stable source for mesenchymal stem cels, which is becoming a cellsource in the regenerative medicine that attracts widespread attentions. OBJECTIVE: To compare the biological characters of mesenchymal stem cels that separated from different components of human placenta (human chorion and vilous trophoblast). METHODS:The amniotic membrane of placenta surface was detached surgicaly. Human chorion and vilous trophoblast in the fetal side was cut into pieces. After digestion with PBS containing colagenase II, mononuclear cels were separated and cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum in 37℃ and 5% CO2, 95% saturated humidity, after 48 hours the ful amount in liquid, dislodge suspension cels. Forty-eight hours later, the medium was changed completed, and non-adherent cels were removed. When cellfusion reached about 90%, trypsin digestion was employed for cellpassage. Biological characters of mesenchymal stem cels separated from different components of human placenta were compared through observation of total number of primary cels, cellmorphology, and surface markers expression (CD90, CD73 and CD105). RESULTS AND CONCLUSION:The flow cytometric analysis revealed that the cels separated from the human chorion and vilous trophoblast were over 90% strongly positive for CD90, CD73, CD105. These two sources of cels showed typical fibroblast morphology, suggesting that the cels have the characteristics of mesenchymal stem cels. Under the same enzyme digestion time, enzyme concentration, and shaking speed, more cels are visible from the chorion, and the subsequent culture is easier to harvest cels.
