Effects of Activin A on paraxial mesodermal differentiation of mouse iPS cells.

<p>(<b>A</b>) A scheme for paraxial mesodermal differentiation of mouse iPS cells with different concentrations of Activin A from day 0 (D0) to day 3 (D3). Activin A was administrated from D0 to D3 at a concentration of 0–20 ng/ml. The cultures also contained BMP4 (10 ng/ml), IGF-1 (10 ng/ml), LiCl (5 mM), and Shh (10 ng/ml). The cells were analyzed on day 6 (D6) in (<b>B</b>), (<b>E</b>), and (<b>F</b>), or on day 1 (D1) in (<b>C</b>) and (<b>D</b>). (<b>B</b>) Total number of mouse iPS cells after differentiation with the protocol shown in (<b>A</b>) (n = 3). (<b>C</b>) Proliferation of differentiated mouse iPS cells on D1 assessed by BrdU assay (n = 3). (<b>D</b>) Apoptosis of differentiated mouse iPS cells on D1 assessed by a proportion of Propidium Iodide (PI) positive/AnnexinV positive cell (n = 3). (<b>E</b>) Dose-dependent induction of PDGFR-α by Activin A in mouse iPS cell differentiation culture. The percentage indicates the proportion of PDGFR-α⁺ cells (n = 3). (<b>F</b>) Gene expression profiles of PDGFR-α⁺ cells in Activin A-induced cultures (n = 3). The expression level of <i>Tbx6</i> and <i>Mesp2</i> genes was reduced in a dose-dependent manner. *p<0.05, **p<0.01 between selected two samples.