[Experimental study of the isolation, culture and in chondrogenic differentiation of human bone mesenchymal stem cell].
OBJECTIVE
To study the isolation of human bone marrow mesenchymal stem cells (MSCs) and in vitro differentiation into chondrocytes as potential seed cell for condyle cartilage tissue engineering.


METHODS
Human MSCs were isolated by percoll solution from normal human bone marrow sample and cultured in flasks. Specific cell surface markers were identified by flow-cytometry. After the cells were treated with inductive medium containing insulin, transferrin, pyruvate, dexathemesone and TGF-beta for 7 - 14 days, microscopic, histological and immuno-histo-chemical studies were performed for chondrogenic phenotype identification.


RESULTS
Primary cultures of human MSCs express CD29 and CD44 positively and meanly, but CD34, CD45 and HLA-DR negatively. After 14 days of induction, the cells were positively stained by safranin O. Immunohistochemical analysis proved strong type II collagen expression.


CONCLUSIONS
Percoll helps to generate a better isolation of MSCs from human bone marrow aspirates with a purity more above 95%. The isolated MSCs can be expanded and induced in vitro to differentiate into chondrocytes by inductive medium.
Guo
Xi-min
XM
Tissue Engineering Research Center, Institute of Basic Medical Science, Academy of Military Medical Sciences, Beijing 100850, China.
Wang
Chang-yong
CY
Wang
Yong-hong
YH
Duan
Cui-mi
CM
Zhao
Qiang
Q
Sun
Da-ming
DM
D001854:Bone Marrow Cells / Q000166:cytology*; D002358:Cartilage, Articular / Q000737:chemistry / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000737:chemistry / Q000166:cytology*; D024043:Collagen Type II / Q000032:analysis; D003907:Dexamethasone / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000494:pharmacology; D008648:Mesoderm / Q000166:cytology*; D011773:Pyruvates / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods; D014168:Transferrin / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D024043:Collagen Type II; D007328:Insulin; D011773:Pyruvates; D014168:Transferrin; D016212:Transforming Growth Factor beta; D003907:Dexamethasone