p130 And pRb in the Maintenance of Transient Quiescence of Mesenchymal Stem Cells

Figure 1

MSCs of the 10T1/2 cell line show serum-dependent growth restriction and cell cycle-dependent oscillation. (a) 10T1/2 cell line possesses the ability to adipocyte differentiation. The cells were observed before AD (i) and in 10 days (ii) after the induction of AD and Red Oil staining under the Pascal microscope (Carl Zeiss, Germany) with transmitted light using a 10x objective. (b) Growth curves of T98G, 10T1/2, and HeLa cell lines: i—full growth medium, ii—serum-deprived medium. Equal cell numbers of each line were plated into wells of 96 cell culture plate in triplicate and counted every day until day 4. The data are presented as , — compared to the 10T1/2 cells, — compared to the T98G cells. (c) Cell proliferation rate of the 10T1/2 cells: i—full growth medium, ii—serum-deprived medium. The cell proliferation rate was estimated using the MTT assay. The asynchronously growing 10T1/2, T98G, and HeLa cells were seeded at a concentration of cells/well in 100 μl full culture medium in 24 wells of 96-well cell culture plate and incubated for 24 h. Then, 12 wells containing cells of each cell line were filled with full growth medium (FM) while the other 12—with serum-free medium (SFM). For the next four days, the cells were treated with the MTT reagent followed by measuring OD by ELISA reader at 570 nm. The data are presented as , — compared to the 10T1/2 cells, — compared to the T98G cells. (d–f) Flow cytometry analysis of cell cycle progression of T98G, MSCs, and HeLa cell lines, accordingly. The cells were cultured for 72 h in DMEM with 0.15% FCS, restimulated with 10% FCS, and used for flow cytometry in 6 h intervals after restimulation.

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