Abstract
Mature mammalian hearts possess very limited regenerative potential. The irreversible cardiomyocyte loss after heart injury can lead to heart failure and death. Pluripotent stem cells (PSCs) can differentiate into cardiomyocytes for cardiac repair, but there are obstacles to their clinical application. Among these obstacles is their potential for post-transplant rejection. Although human amniotic fluid-derived stem cells (hAFSCs) are immune privileged, they cannot induce cardiac differentiation. Thus, we generated hAFSC-derived induced PSCs (hAFSC-iPSCs) and used a Wnt-modulating differentiation protocol for the cardiac differentiation of hAFSC-iPSCs. In vitro studies using flow cytometry, immunofluorescence staining, and patch-clamp electrophysiological study, were performed to identify the characteristics of hAFSC-iPSC-derived cardiomyocytes (hAFSC-iPSC-CMs). We injected hAFSC-iPSC-CMs intramuscularly into rat infarcted hearts to evaluate the therapeutic potential of hAFSC-iPSC-CM transplantation. At day 21 of differentiation, the hAFSC-iPSC-CMs expressed cardiac-specific marker (cardiac troponin T), presented cardiomyocyte-specific electrophysiological properties, and contracted spontaneously. Importantly, these hAFSC-iPSC-CMs demonstrated low major histocompatibility complex (MHC) class I antigen expression and the absence of MHC class II antigens, indicating their low immunogenicity. The intramyocardial transplantation of hAFSC-iPSC-CMs restored cardiac function, partially remuscularized the injured region, and reduced fibrosis in the rat infarcted hearts. Therefore, hAFSC-iPSCs are potential candidates for the repair of infarcted myocardium.
Highlights
Human pluripotent stem cells, i.e., human embryonic stem cells and human induced pluripotent stem cells, can differentiate into functionally contracting cardiomyocytes using a well-established Wnt-signaling modulating directed cardiac differentiation protocol [1,2,3,4]
Characterization of Human pluripotent stem cells (hPSCs) with and without Cardiac Differentiation human amniotic fluid-derived stem cells (hAFSCs)-induced pluripotent stem cells (iPSCs) and human embryonic stem cells (hESCs) were expanded using α-Minimum Essential Media (MEM) supplemented with 4 ng/mL of basic fibroblast growth factor (233-FB; R&D Systems, Minneapolis, MN, USA)
When these cultured cells reached confluence, we used a serum-free, monolayer direct cardiac differentiation protocol involving a serial application of activin A and bone morphogenetic protein-4 (Figure 1a) [1,2,3,4,16,17] to differentiate both hAFSC-iPSCs and hESCs into cardiomyocytes
Summary
Human pluripotent stem cells (hPSCs), i.e., human embryonic stem cells (hESCs) and human induced pluripotent stem cells (iPSCs), can differentiate into functionally contracting cardiomyocytes using a well-established Wnt-signaling modulating directed cardiac differentiation protocol [1,2,3,4]. The recipients have to rely on taking strong lifelong immunosuppressive agents to prevent rejection of these transplanted cells. Human amniotic fluid-derived stem cells (hAFSCs) possess the favorable property of pluripotency [13,14,15] and the immune privilege due to low major histocompatibility complex (MHC) class I antigen expression and the absence of MHC class II antigens [13,14,15,16], we demonstrate that hAFSCs cannot differentiate into functional cardiomyocytes in vitro using the Wnt-signaling modulating directed cardiac differentiation protocol [17]. Using a relatively less potent immunosuppressive therapy, the engrafted hAFSC-iPSC-CMs could survive in the infarcted area of the rat hearts and restore the post-MI cardiac function. HAFSC-iPSCs are potential, ideal candidates for cell therapy
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