File S1 - Molecular Imaging of Induced Pluripotent Stem Cell Immunogenicity with In Vivo Development in Ischemic Myocardium

Table S1. Primers used in the PCR. Table S2. Infiltrating inflammatory cells in MI mice receiving PBS injection (n = 4 for each detection time). Figure S1. Characterization and Comparison between control iPS and iPS-TF cells. Both iPS and iPS-TF colonies cells were positive for pluripotent ESC markers – SOX2, SSEA-1, NANOG and OCT4. In iPS-TF colonies, colocalization of mrfp and ESC markers were observed under a fluorescent microscope, while in iPS colonies, no expression of mrfp was observed. The nucleus were stained with hochest (Bar = 100 µm) (A). The expression of ESC markers were also analyzed in gene level by RT-PCR, no significant difference were observed between the control iPS and iPS-TF cells(B-a, B-b); Cell viabilities and proliferation of iPS and iPS-TF cells were analyzed and compared within 72 h culture. Quantification of viable cells at 24 h, 48 h and 72 h time points showed no significant difference between iPS and iPS-TF cells(B-c); No significant difference in proliferation capacities was observed between iPS and iPS-TF cells either(B-d); After EB formation and differentiation, both iPS and iPS-TF cells could differentiate into tridermic lineages(B-e). Figure S2. Stable expression of repoter gene in iPSC-TF cells. (A), Stable expression of repoter gene during long-term passage of iPSCs; (B, C), Bioluminescence assay on the same number (105) of iPSC-TF cells from different passages. Figure S3. Differentiation and characterization of iPSC-CMs. (A), Comparison of the contracting areas (Bar = 100 µm). (C) The expression of cardiac genes in Vc-induced/spontaneous differentiation of iPSCs; (D) Contracting EBs were positively stained by cardiac-specfic antibodies. (Bar = 50 µm). (E) Expression of reporters in iPSC-noncontracting and contracting derivates; (F), Expression of reporters in iPSC-CMs(Bar = 50 µm); (G) The expression of pluripotent, cardiac genes in iPSCs, iPSC-cardiac derivates and iPSC-CMs. Lane 1: iPSCs; Lane 2: iPSC-cardiac derivates; Lane 3: iPSC-CMs; (H) Comparison of iPSCs, iPSC-cardiac derivates and iPSC-CMs. *pSupplementary Methods. Production of lentiviral vectors carrying tri-fusion reporter gene and establishment of iPS-TF line. (DOC)