Cardiomyocyte-mediated contact programs human mesenchymal stem cells to express cardiogenic phenotype Sunil Rangappa, MD, John W.C Entwistle, PhD, MD,

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Presentation transcript:

Cardiomyocyte-mediated contact programs human mesenchymal stem cells to express cardiogenic phenotype Sunil Rangappa, MD, John W.C Entwistle, PhD, MD, Andrew S Wechsler, MD, J.Yasha Kresh, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 126, Issue 1, Pages 124-132 (July 2003) DOI: 10.1016/S0022-5223(03)00074-6

Figure 1 A, Group I cardiomyocytes grown in the tissue culture plates. Group II conditioned media prepared from the cardiomyocytes after 48 hours in culture. Group III coculture of hMSC with cardiomyocytes. B, Conditioned media prepared from the soluble factors released from the cardiomyocytes, in which hMSC were subsequently cultured. The Journal of Thoracic and Cardiovascular Surgery 2003 126, 124-132DOI: (10.1016/S0022-5223(03)00074-6)

Figure 2 a, Spindle-shaped human mesenchymal stem cells (20×), 24 hours after seeding on cell polystyrene cell culture surface. b, Rod-shaped human cardiomyocytes forming intercellular junctions with neighboring cells. The Journal of Thoracic and Cardiovascular Surgery 2003 126, 124-132DOI: (10.1016/S0022-5223(03)00074-6)

Figure 3 a to h, Left panel shows positive controls, consisting of cardiomyocytes. Right panel shows transformed hMSCs (10×). a, No staining observed against fast myosin (MF-1) in human cardiomyocytes. b, No staining observed against fast myosin (MF-1) in untreated hMSCs (negative control). d, Positive staining for sarcomeric myosin (MF-20) in cocultured hMSCs. f, Positive staining for heavy chain myosin (β-MyHC) in cocultured hMSCs. h, Positive staining for troponin-T in cocultured hMSCs. j, Negative staining against troponin-I. c, e, g, i, Positive staining in cultured cardiomyocytes for MF-20, β-MyHC, troponin-T, and troponin-I, respectively. The Journal of Thoracic and Cardiovascular Surgery 2003 126, 124-132DOI: (10.1016/S0022-5223(03)00074-6)

Figure 4 RT-PCR for specific hMSC transformation markers (48-hour coculture). Lane 1: 1-kb DNA ladder; lane 2: hMSC (negative control); lane 3: conditioned media treated with hMSC in the presence of serum; lane 4 conditioned media treated with hMSC in the absence of serum; lane 5: cell-cell contact-dependent coculture; lane 6: human cardiomyocytes (positive control). β-MHC, β-actin, and CT-3 expression in cell-cell contact dependent coculture. Expression of β-actin in conditioned media ± serum. GADPH is internal control. Tn-I is troponin-I expression in cultured cardiomyocytes. The Journal of Thoracic and Cardiovascular Surgery 2003 126, 124-132DOI: (10.1016/S0022-5223(03)00074-6)

Figure 5 Expression of gap junctions connexin-specific genes (Cx-32, Cx-40, Cx-43, and Cx-45) in hMSC. Lane 1: 1-kb DNA ladder; lane 2: hMSC expression of specific connexins; lane 3: expression of connexin-specific genes in human cardiomyocytes (positive control). The Journal of Thoracic and Cardiovascular Surgery 2003 126, 124-132DOI: (10.1016/S0022-5223(03)00074-6)