Amniotic fluid-derived stem cells demonstrate limited cardiac differentiation following small molecule-based modulation of Wnt signaling pathway

Connell, Jennifer P.; Ruano, Rodrigo; Jacot, Jeffrey G.

doi:10.1088/1748-6041/10/3/034103

Cited by 13 publications

(18 citation statements)

References 31 publications

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“…In our study, the isolated cells from amniotic fluid had a phenotype that was consistent with that of MSCs [ 8 , 57 ]. Because these AF‐MSCs had an intermediate phenotype between that of true pluripotent stem cells and fully differentiated adult stromal cells [ 58 – 60 ], our amniocytes should not be confused with CD117 + AFSCs [ 37 ]. Although both AF‐MSCs and AFSCs proliferate easily in culture, our group, among others, has shown that AF‐MSCs undergo senescence following repetitive passaging, are not pluripotent, and cannot be differentiated directly into CMs using established protocols [ 60 ].…”

Section: Discussionmentioning

confidence: 98%

“…Because these AF‐MSCs had an intermediate phenotype between that of true pluripotent stem cells and fully differentiated adult stromal cells [ 58 – 60 ], our amniocytes should not be confused with CD117 + AFSCs [ 37 ]. Although both AF‐MSCs and AFSCs proliferate easily in culture, our group, among others, has shown that AF‐MSCs undergo senescence following repetitive passaging, are not pluripotent, and cannot be differentiated directly into CMs using established protocols [ 60 ]. In contrast, AFSCs have been elegantly shown to be pluripotent on the basis of a transgene‐free, chemical‐based approach using the histone deacetylase inhibitor, valproic acid [ 61 ].…”

Section: Discussionmentioning

confidence: 99%

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Human Cardiomyocytes Prior to Birth by Integration-Free Reprogramming of Amniotic Fluid Cells

Jiang

Herron

Bernardo

et al. 2016

Stem Cells Translational Medicine

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The establishment of an abundant source of autologous cardiac progenitor cells would represent a major advance toward eventual clinical translation of regenerative medicine strategies in children with prenatally diagnosed congenital heart disease. In support of this concept, we sought to examine whether functional, transgene-free human cardiomyocytes (CMs) with potential for patient-specific and autologous applications could be reliably generated following routine amniocentesis. Under institutional review board approval, amniotic fluid specimens (8-10 ml) at 20 weeks gestation were expanded and reprogrammed toward pluripotency using nonintegrating Sendai virus (SeV) expressing OCT4, SOX2, cMYC, and KLF4. Following exposure of these induced pluripotent stem cells to cardiogenic differentiation conditions, spontaneously beating amniotic fluid-derived cardiomyocytes (AF-CMs) were successfully generated with high efficiency. After 6 weeks, quantitative gene expression revealed a mixed population of differentiated atrial, ventricular, and nodal AF-CMs, as demonstrated by upregulation of multiple cardiac markers, including MYH6, MYL7, TNNT2, TTN, and HCN4, which were comparable to levels expressed by neonatal dermal fibroblast-derived CM controls. AF-CMs had a normal karyotype and demonstrated loss of NANOG, OCT4, and the SeV transgene. Functional characterization of SIRPA + AF-CMs showed a higher spontaneous beat frequency in comparison with dermal fibroblast controls but revealed normal calcium transients and appropriate chronotropic responses after b-adrenergic agonist stimulation. Taken together, these data suggest that somatic cells present within human amniotic fluid can be used to generate a highly scalable source of functional, transgene-free, autologous CMs before a child is born. This approach may be ideally suited for patients with prenatally diagnosed cardiac anomalies. STEM CELLS TRANSLATIONAL MEDICINE 2016;5:1595-1606 SIGNIFICANCEThis study presents transgene-free human amniotic fluid-derived cardiomyocytes (AF-CMs) for potential therapy in tissue engineering and regenerative medicine applications. Using 8-10 ml of amniotic fluid harvested at 20 weeks gestation from normal pregnancies, a mixed population of atrial, ventricular, and nodal AF-CMs were reliably generated after Sendai virus reprogramming toward pluripotency. Functional characterization of purified populations of beating AF-CMs revealed normal calcium transients and appropriate chronotropic responses after b-adrenergic agonist stimulation in comparison with dermal fibroblast controls. Because AF-CMs can be generated in fewer than 16 weeks, this approach may be ideally suited for eventual clinical translation at birth in children with prenatally diagnosed cardiac anomalies.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Human Cardiomyocytes Prior to Birth by Integration-Free Reprogramming of Amniotic Fluid Cells

Jiang

Herron

Bernardo

et al. 2016

Stem Cells Translational Medicine

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“…AFSC were isolated based on previously published methods from our group[8, 13]. Primary human amniotic fluid was obtained from patients in their second trimester undergoing planned amnioreduction as part of a therapeutic treatment for twin-twin transfusion syndrome (TTTS).…”

Section: Methodsmentioning

confidence: 99%

“…Specifically, AFSC express mesenchymal stem cell markers (CD29, CD44, CD90, and CD105), certain pluripotent markers (SOX2), and are capable of differentiating into all three germ layers[8]. While attempts at direct cardiac differentiation have shown gene and protein level similarities (GATA4, Nkx2.5, α-actinin, cTnT), resulting cells ultimately lack contractility[8, 9]. …”

Section: Introductionmentioning

confidence: 99%

Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells

et al. 2017

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Congenital heart defects are the most common birth defect. The limiting factor in tissue engineering repair strategies is an autologous source of functional cardiomyocytes. Amniotic fluid contains an ideal cell source for prenatal harvest and use in correction of congenital heart defects. This study aims to investigate the potential of amniotic fluid-derived stem cells (AFSC) to undergo non-viral reprogramming into induced pluripotent stem cells (iPSC) followed by growth-factor-free differentiation into functional cardiomyocytes. AFSC from human second trimester amniotic fluid were transfected by non-viral vesicle fusion with modified mRNA of OCT4, KLF4, SOX2, LIN28, cMYC and nuclear GFP over 18 days, then differentiated using inhibitors of GSK3 followed 48 hours later by inhibition of WNT. AFSC-derived iPSC had high expression of OCT4, NANOG, TRA-1-60, and TRA-1-81 after 18 days of mRNA transfection and formed teratomas containing mesodermal, ectodermal, and endodermal germ layers in immunodeficient mice. By Day 30 of cardiomyocyte differentiation, cells contracted spontaneously, expressed connexin 43 and β-myosin heavy chain organized in sarcomeric banding patterns, expressed cardiac troponin T and β-myosin heavy chain, showed upregulation of NKX2.5, ISL-1 and cardiac troponin T with downregulation of POU5F1, and displayed calcium and voltage transients similar to those in developing cardiomyocytes. These results demonstrate that cells from human amniotic fluid can be differentiated through a pluripotent state into functional cardiomyocytes.

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“…The special section combines original research papers and review articles that present recent progress in development of cardiac tissue engineering biomaterials. It discusses potential cell sources in original papers [4, 5] and reviews [6]. The use of various hydrogels for cardiovascular tissue engineering is reviewed [7, 8] and original papers on development of brand new scaffold materials that incorporate architectural complexity of the native myocardium are presented [9, 10].…”

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confidence: 99%

Biomaterials for cardiac tissue engineering

Radišić

2015

Biomed. Mater.

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Amniotic fluid-derived stem cells demonstrate limited cardiac differentiation following small molecule-based modulation of Wnt signaling pathway

Cited by 13 publications

References 31 publications

Human Cardiomyocytes Prior to Birth by Integration-Free Reprogramming of Amniotic Fluid Cells

Human Cardiomyocytes Prior to Birth by Integration-Free Reprogramming of Amniotic Fluid Cells

Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells

Biomaterials for cardiac tissue engineering

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