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. 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.