Evaluation of human MSCs cell cycle, viability and differentiation in micromass culture.
Mesenchymal stem cells (MSCs) have the potential to differentiate into distinct mesenchymal tissue cells. They are easy to expand while maintaining their undifferentiated state, which suggests that these cells could be an attractive cell source for tissue engineering of cartilage. In vitro high density micromass culture has been widely used for chondrogenesis induction. Our objective was to investigate human MSCs cell cycle, viability and differentiation in these conditions. Therefore, to induce human MSCs chondrogenesis, micromasses were cultured in the presence of transforming growth factor-beta1 in serum free medium for 21 days. Cell cycle, cell viability and cell phenotype were analyzed by flow cytometry. From day 0 to 7, the G0/G1 phase increased, whereas the S phase decreased gradually, but cell cycle phases (S, G0/G1 and G2/M) did not significantly change after day 7. Less than 10% of cells were apoptotic, but no necrosis was observed, even at day 21. We observed a decrease in CD90 and CD105 expression, from day 0 to 21. In conclusion, our results demonstrate a good viability of human MSCs in micromass culture during the whole period of culture. Moreover, micromass culture allowed human MSCs to be synchronized at the G0/G1 phase, while their phenotype suggested some degree of differentiation.
Yang
Jing-Wei
JW
Research Group on Cell and Tissue Engineering and Mechanics, LEMTA UMR CNRS 7563, Faculty of Medicine, UHP Nancy 1, Vandoeuvre-lès-Nancy, France.
de Isla
Natalia
N
Huselstein
Céline
C
Sarda-Kolopp
Marie-Nathalie
MN
Li
Na
N
Li
Yin-Ping
YP
Jing-Ping
Ou-Yang
OY
Stoltz
Jean-François
JF
Eljaafari
Assia
A
D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D020219:Chondrogenesis / Q000502:physiology*; D000071063:Endoglin; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D018800:Thy-1 Antigens / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D011956:Receptors, Cell Surface; C508960:TGFB1 protein, human; D018800:Thy-1 Antigens; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1