[Biologic characteristics of rat bone marrow mesenchymal stem cells cultured in vitro].
OBJECTIVE
To investigate biological characteristics of rat bone marrow mesenchymal stem cells (MSC) cultured in vitro and to explore their potential applications.


METHODS
MSC were isolated from rat bone marrow by density gradient centrifugation and were induced to differentiation. Flow cytometry was used to characterize their surface antigen expression, cell cycle status and cell growth parameters. Telomerase activity was determined by TRAP-ELISA assay.


RESULTS
Fusiform MSC became larger and flattener with increasing passages of culture. After the fourth passage, the MSC showed an immunophenotype of CD29 (94.75% +/- 3.68%), CD71 (95.43% +/- 2.23%), and CD90 (98.08% +/- 3.88%). After the seventh passage, MSC with such immunophenotype decreased with CD29: 50.00% +/- 3.35%, CD71: 50.70% +/- 2.43%, and CD90: 48.60% +/- 2.83%. Cells with such immunoprofile completely disappeared after passage 9. Overall, MSC grew faster during the first 5 passages. The number of MSC in S and G(2)/M phases were 38.36% +/- 2.01% and those in G(0)/G(1) phase were 61.64% +/- 2.13% after 3 passages. The cell growth decreased after passage 7. Percentage of MSCs in S and G(2)/M phases was 10.83% +/- 1.63% and that in G(0)/G(1) was 89.17% +/- 1.96% after passage 12, after which the cells failed to further divide. After passage 9, MSCs lost their ability to differentiate to Von Kossa and oil red O positive staining cells. In addition, telomerase activity of MSC also gradually decreased with the prolonged passages, from the original 52.7% +/- 0.78% to no telomerase activity.


CONCLUSIONS
The biological and immunophenotypical characteristics of cultured MSC showed obvious alterations with increasing numbers of passage of culture.
Yu
Xiao-xia
XX
The Key Laboratory of Ministry of Education, Department of Pathobiology, Jilin University, Changchun 130021, China.
Shi
Ying-ai
YA
Xin
Ying
Y
Zhang
Li-hong
LH
Li
Yu-lin
YL
Wu
Shan
S
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002453:Cell Cycle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D016130:Immunophenotyping; D019012:Integrin beta1 / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D011990:Receptors, Transferrin / Q000378:metabolism; D018800:Thy-1 Antigens / Q000378:metabolism
D015703:Antigens, CD; C068126:CD71 antigen; D019012:Integrin beta1; D011990:Receptors, Transferrin; D018800:Thy-1 Antigens