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Amniotic membrane as novel scaffold for human iPSC-derived cardiomyogenesis.
In Vitro Cell Dev Biol Anim. 2019 Apr; 55(4):272-284.VC

Abstract

Recent approaches of using decellularized organ matrices for cardiac tissue engineering prompted us to culture human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) on the human amniotic membrane (hAM). Since hAM has been used lately to patch diseased hearts in patients and has shown anti-inflammatory and anti-fibrotic benefits, it qualifies as a cardiac compatible and clinically relevant heart tissue scaffold. The aim of this study was to test the ability of the hAM to support attachment, differentiation, and maturation of hiPSC-derived CMs in vitro. hAMs were prepared from term placenta. An in-house generated hiPSC line was used for CM derivation. hiPSC-derived cardiac progenitors were cultured on the surface of cryopreserved hAMs and in the presence of cytokines promoting cardiac differentiation. CMs grown on hAM and popular basement membrane matrix (BMM) Matrigel™ were compared for the following aspects of cardiac development: the morphology of cardiomyocytes with respect to shape and cellular alignments, levels of cardiac-related gene transcript expression, functionality in terms of spontaneous calcium fluxes and mitochondrial densities and distributions. hAM is biocompatible with hiPSC-derived CMs. hAM increased cardiac transcription regulator and myofibril protein transcript expression, accelerated intracellular calcium transients, and enhanced cellular mitochondrial complexity of its cardiomyocytes in comparison to cardiomyocytes differentiated on Matrigel™. Our data suggests that hAM supports differentiation and improves cardiomyogenesis in comparison to Matrigel™. hAMs are natural, easily and largely available. The method of preparing hAM cardiac sheets described here is simple with potential for clinical transplantation. Graphical abstract A An outline of the differentiation protocol with stage-specific growth factors and culture media used. B Cell fates from pluripotent stem cells to cardiomyocytes during differentiation on the amniotic membrane. C-FPhotomicrographs of cells at various stages of differentiation. Scale bars represent 100 μm.

Authors+Show Affiliations

Manipal Academy of Higher Education, School of Regenerative Medicine, GKVK post, Bellary Road, Bengaluru, Karnataka, 560065, India. shagufta.parveen@manipal.edu.National Centre for Biological Sciences, TIFR, Bangalore, India.National Centre for Biological Sciences, TIFR, Bangalore, India.Stempeutics Research Pvt. Ltd., Bangalore, India.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30798515

Citation

Parveen, Shagufta, et al. "Amniotic Membrane as Novel Scaffold for Human iPSC-derived Cardiomyogenesis." In Vitro Cellular & Developmental Biology. Animal, vol. 55, no. 4, 2019, pp. 272-284.
Parveen S, Singh SP, Panicker MM, et al. Amniotic membrane as novel scaffold for human iPSC-derived cardiomyogenesis. In Vitro Cell Dev Biol Anim. 2019;55(4):272-284.
Parveen, S., Singh, S. P., Panicker, M. M., & Gupta, P. K. (2019). Amniotic membrane as novel scaffold for human iPSC-derived cardiomyogenesis. In Vitro Cellular & Developmental Biology. Animal, 55(4), 272-284. https://doi.org/10.1007/s11626-019-00321-y
Parveen S, et al. Amniotic Membrane as Novel Scaffold for Human iPSC-derived Cardiomyogenesis. In Vitro Cell Dev Biol Anim. 2019;55(4):272-284. PubMed PMID: 30798515.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Amniotic membrane as novel scaffold for human iPSC-derived cardiomyogenesis. AU - Parveen,Shagufta, AU - Singh,Shishu Pal, AU - Panicker,M M, AU - Gupta,Pawan Kumar, Y1 - 2019/02/24/ PY - 2018/08/28/received PY - 2019/01/08/accepted PY - 2019/2/25/pubmed PY - 2019/7/20/medline PY - 2019/2/25/entrez KW - Human amniotic membrane KW - Improved cardiomyogenesis KW - Placental induced pluripotent stem cells KW - hAM cardiac sheets KW - hiPSC-derived cardiomyocytes SP - 272 EP - 284 JF - In vitro cellular & developmental biology. Animal JO - In Vitro Cell. Dev. Biol. Anim. VL - 55 IS - 4 N2 - Recent approaches of using decellularized organ matrices for cardiac tissue engineering prompted us to culture human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) on the human amniotic membrane (hAM). Since hAM has been used lately to patch diseased hearts in patients and has shown anti-inflammatory and anti-fibrotic benefits, it qualifies as a cardiac compatible and clinically relevant heart tissue scaffold. The aim of this study was to test the ability of the hAM to support attachment, differentiation, and maturation of hiPSC-derived CMs in vitro. hAMs were prepared from term placenta. An in-house generated hiPSC line was used for CM derivation. hiPSC-derived cardiac progenitors were cultured on the surface of cryopreserved hAMs and in the presence of cytokines promoting cardiac differentiation. CMs grown on hAM and popular basement membrane matrix (BMM) Matrigel™ were compared for the following aspects of cardiac development: the morphology of cardiomyocytes with respect to shape and cellular alignments, levels of cardiac-related gene transcript expression, functionality in terms of spontaneous calcium fluxes and mitochondrial densities and distributions. hAM is biocompatible with hiPSC-derived CMs. hAM increased cardiac transcription regulator and myofibril protein transcript expression, accelerated intracellular calcium transients, and enhanced cellular mitochondrial complexity of its cardiomyocytes in comparison to cardiomyocytes differentiated on Matrigel™. Our data suggests that hAM supports differentiation and improves cardiomyogenesis in comparison to Matrigel™. hAMs are natural, easily and largely available. The method of preparing hAM cardiac sheets described here is simple with potential for clinical transplantation. Graphical abstract A An outline of the differentiation protocol with stage-specific growth factors and culture media used. B Cell fates from pluripotent stem cells to cardiomyocytes during differentiation on the amniotic membrane. C-FPhotomicrographs of cells at various stages of differentiation. Scale bars represent 100 μm. SN - 1543-706X UR - https://www.unboundmedicine.com/medline/citation/30798515/Amniotic_membrane_as_novel_scaffold_for_human_iPSC_derived_cardiomyogenesis_ L2 - https://dx.doi.org/10.1007/s11626-019-00321-y DB - PRIME DP - Unbound Medicine ER -