[Study on chondrogenesis of rabbit bone marrow-derived mesenchymal stem cells by centrifuge-tube culture in vitro].
OBJECTIVE
To investigate the feasibility of constructing tissue engineered cartilage by inducing bone marrow-derived mesenchymal stem cells (MSCs) in centrifuge-tube culture.


METHODS
By Percoll density gradient centrifuge and monolaryer culture in vitro MSCs were isolated from rabbit bone marrow and were purified. Some MSCs were seeded into centrifuge tube culture and allowed to form three-dimensional aggregates in a chemically defined medium mainly including transforming growth factor- b1 (TGF-beta1, 10 ng/ml), dexamethasone (10-7), ascorbic acid (50 microg/ml), 1% insulin, 1% selenium and 1% transferring. At day 7, 14, 21, aggregates were take out and specific markers of differentiation of cartilage were detected by the appearance of toluiding blue merachromasia and the immunohistochemistry detection of type II collagen and in situ hybridization detection of collagen type II mRNA.


RESULTS
MSCs seeded into centrifuge tube culture became analogous cartilage tissue after 7-21 days, their specific markers of chondrogeneses were positive in the appearance of toluiding blue merachromasia and the immunohistochemistry detection of type II collagen and in situ hybridization detection of collagen type II mRNA.


CONCLUSIONS
MSCs have the capacity to undergo chondrogenic differentiation.
Huang
Dingqiazng
D
Department of Otolaryngology, the Affiliated Hospital of Luzhou Medical College, Luzhou, 646000, China. hdingq1971@tom.com
Liang
Chuanyu
C
Wang
Lihong
L
Liu
Shixi
S
Li
Shengfu
S
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D020219:Chondrogenesis; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011817:Rabbits