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(a,b) Histologic analysis of graft cell survival. Heat-shocked hES cell–derived cardiomyocytes were injected into infarcted hearts of nude rats in the presence of SFM, Matrigel-only (a, Cells+Matrigel), or the full pro-survival cocktail (PSC) including Matrigel (b, Cells+PSC) (n = 5 per group). Sections were stained with an antibody to 
-myosin heavy chain ( MHC, red chromagen) as well as a human-specific pan-centromeric in situ hybridization probe (huCent, brown DAB deposit) to identify total human (that is, huCent+) and specifically human cardiac (that is, MHC and huCent double-positive) graft cells. The human cardiomyocytes, indicated by arrows, were significantly more abundant in histologic sections from the Cells+PSC group than in Cells+Matrigel alone group. Histology is not depicted from the recipients of cells in SFM alone because none of these hearts showed even a single surviving human nucleus after 1 week. Counterstain, fast green; scale bar, 50 
m. (c) Quantification of hES cell–derived cardiomyocyte graft size. Although no grafts were detected in any rats receiving hES cell–derived cardiomyocytes delivered in SFM alone (Cells+SFM), all rats receiving cells delivered in Matrigel-only (Cells+Matrigel) or in the full prosurvival cocktail (Cells+PSC) showed surviving graft (5/5 rats per group). However, recipients of cells in the full prosurvival cocktail (Cells+PSC) showed a mean of approximately fourfold more -myosin-positive graft cells than did the Matrigel-only group. Note that counts indicate the total number of cells observed on sampled sections, not the total number of cells per heart. *, P < 0.05.
