Global gene expression of mouse iPSC directed differentation towards smooth muscle cells using an Acta2hrGFP mouse iPSC reporter line. Global gene expression of mouse iPSC directed differentation towards smooth muscle cells using an Acta2hrGFP mouse iPSC reporter line

Here, we generated and differentiated a mouse induced pluripotent stem cell line with an Acta2hrGFP reporter towards a smooth muscle-like cell that can be purified and expresses characteristic markers of smooth muscle cells. We performed microarray analysis of 3 timepoints in our smooth muscle directed differentiation protocol and compared it to primary adult aortic smooth muscle cells. The profiles indicated that the day 13 Acta2hrGFP+ and Acta2hrGFP- populations are fairly similar, and that the day13 Acta2hrGFP+ population's transcriptomic profile is reminiscent of an immature or a synthetic smooth muscle cell phenotype. Using a mouse iPSC line with a transgenic GFP reporter for Acta2, we characterized the global transcriptomic proifle of cells during different developmental time points in our in vitro differentiation (undifferentiated iPSC, sorted Kdr+ mesodermal progenitors, and sorted Acta2hrGFP+ and Acta2hrGFP- candidate SMCS). We also included freshly isolated primary aortic Acta2hrGFP+ smooth muscle cells as a control. Overall design: We prepared microarray expression profiles representing the following 3 stages of iPSC-SMC directed differentiation: undifferentiated iPSC (day 0), sorted Kdr+ mesodermal progenitors (day 5), and sorted Acta2hrGFP+ and Acta2hrGFP- candidate SMCs (day 13). For positive controls we included primary mouse aortic Acta2hrGFP+ SMCs. Biological triplicates of all samples were prepared. Global gene expression in all 15 samples was analyzed by Affymetrix GeneChip Mouse Gene 2.0 ST arrays.