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In Vitro Cell Dev Biol Anim. 2019 Apr;55(4):272-284. doi: 10.1007/s11626-019-00321-y. Epub 2019 Feb 24.

Amniotic membrane as novel scaffold for human iPSC-derived cardiomyogenesis.

Author information

1
Manipal Academy of Higher Education, School of Regenerative Medicine, GKVK post, Bellary Road, Bengaluru, Karnataka, 560065, India. shagufta.parveen@manipal.edu.
2
National Centre for Biological Sciences, TIFR, Bangalore, India.
3
Stempeutics Research Pvt. Ltd., Bangalore, India.

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.

KEYWORDS:

Human amniotic membrane; Improved cardiomyogenesis; Placental induced pluripotent stem cells; hAM cardiac sheets; hiPSC-derived cardiomyocytes

PMID:
30798515
DOI:
10.1007/s11626-019-00321-y

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