Furthermore, one disadvantage of transplanting immature PSC-CMs is arrhythmogenesis after engraftment, which is another limitation of this treatment therefore, at least some promotion of cell maturation is necessary before transplantation. These data suggest that proliferation is not the only factor contributing to hPSC-CM engraftment. Moreover, 3-day-old hPSC-derived cardiovascular progenitor cells could not engraft or survive in infarcted non-human primate (NHP) hearts, whereas 20-day-old PSC-CMs clearly achieved remuscularization in NHP hearts. For example, 8-day-old hPSC-CMs demonstrated lower engraftment efficiency than 20-day-old hPSC-CMs post-transplantation in mouse hearts. Thus, one potential advantage of transplanting immature cardiac progenitor cells is their higher proliferative capacity however, hPSC-CMs younger than 10 days old were shown to be too immature to engraft.
In general, the maturity of hPSC-CMs is inversely correlated with their proliferative capacity. One potential approach to increase the engraftment efficiency is the promotion of cell cycle activity in hPSC-CMs, but this strategy may enhance the risk of tumorigenicity. Although clinical studies have been initiated in several countries, hPSC-CM treatments still have some critical limitations, including poor engraftment of the transplanted CMs. Accumulating evidence has revealed that PSC-CMs can heal injured hearts through not only indirect paracrine effects but also direct remuscularization. This study revealed the benefits of long-term culture, which may enhance the therapeutic potential of hiPSC-CMs.Ĭardiac regenerative therapy using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is expected to be a next-generation therapy for failing hearts because of the unlimited cardiogenic potential of hPSC-CMs. CRYAB, which was highly expressed in D56-CMs, was identified as an angiogenic factor from mature hiPSC-CMs. Long-term cultured mature hiPSC-CMs promoted engraftment, maturation and angiogenesis post-transplantation in infarcted rat hearts. Furthermore, CRYAB overexpression enhanced angiogenesis in the D28-CM grafts at 4 weeks post-transplantation.
Endothelial cell migration was inhibited by small interfering RNA-mediated knockdown of CRYAB when co-cultured with D56-CMs in vitro. As shown by transcriptomic and western blot analyses, the expression of a small heat shock protein, alpha-B crystallin (CRYAB), was significantly upregulated in D56-CMs compared with D28-CMs. Transcriptomic sequencing analysis revealed that compared with the engrafted D28-CMs, the engrafted D56-CMs enriched genes related to blood vessel regulation at 12 weeks post-transplantation. Notably, D56-CMs consistently promoted microvessel formation in the graft area from 1 to 12 weeks post-transplantation. Histological and immunohistochemical analyses showed that D56-CMs promoted engraftment and maturation in the graft area at 12 weeks post-transplantation. In vivo bioluminescence imaging studies revealed increased bioluminescence intensity of D56-CMs at 8 and 12 weeks post-transplantation. Upregulated expression of mature sarcomere genes in vitro was observed in D56-CMs compared with D28-CMs. We performed transcriptomic sequencing analysis to elucidate the genetic profiles before and after hiPSC-CM transplantation. We transplanted D28-CMs or D56-CMs into the hearts of rat myocardial infarction models and examined cell retention and engraftment via in vivo bioluminescence imaging and histological analysis. MethodsĬardiomyocytes were differentiated from hiPSCs via monolayer culturing, and the cells were harvested on day 28 or 56 (D28-CMs or D56-CMs, respectively) after differentiation. We aimed to investigate the benefits of long-term cultured mature hiPSC-CMs in injured rat hearts. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) can be used to treat heart diseases however, the optimal maturity of hiPSC-CMs for effective regenerative medicine remains unclear.
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