Knockdown of RB in Early-Passage MSCs Increased the Cell Size and Induced Loss of the Proliferation Rate and In Vitro Osteogenic and Adipogenic Differentiation Potentials
Early-passage MSCs were infected with lentivirus carrying control (CTR) or RB-specific shRNAs, siRB(1) or siRB(2).
(A) Left: quantitative RT-PCR and western blot analysis of RB knockdown efficiency. Cells were seeded at 103 cells/cm2 and a subculture was performed at 1-week intervals. Middle: cell number fold increase. Right: population doubling time.
(B) Left: morphology of MSCs without or with RB knockdown. Right: cell size measurements.
(C and D) MSCs without or with RB knockdown were treated with OIM. (C) Left: representative pictures of alizarin red S staining at 2 weeks. Right: optical density measurements of extracted alizarin red S staining. (D) Quantitative RT-PCR showed the mRNA levels at 1 week.
(E and F) MSCs without or with RB knockdown were treated with AIM. (E) Left: representative pictures of oil red O staining at 2 weeks. Right: optical density measurements of extracted oil red O staining. (F) Quantitative RT-PCR showed the mRNA levels at 1 week.
The results are expressed as mean ± SD of three independent experiments. Asterisks indicate significant differences (∗∗p < 0.01). Scale bar, 50 μm. See also Figure S2.