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

Aaron J. Velasquez-Mao, Christopher J.M. Tsao, Madeline N. Monroe, Xavier Legras, Beatrice Bissig-Choisat, Karl Dimiter Bissig, Rodrigo Ruano, Jeffrey G. Jacot

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article numbere0177824
JournalPLoS One
Volume12
Issue number5
DOIs
StatePublished - May 1 2017
Externally publishedYes

Fingerprint

amniotic fluid
Amniotic Fluid
Stem cells
Cardiac Myocytes
stem cells
Stem Cells
Fluids
troponin T
Induced Pluripotent Stem Cells
Troponin T
Myosin Heavy Chains
Congenital Heart Defects
myosin heavy chains
Defects
heart
Growth Differentiation Factors
connexins
Germ Layers
tissue engineering
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Velasquez-Mao, A. J., Tsao, C. J. M., Monroe, M. N., Legras, X., Bissig-Choisat, B., Bissig, K. D., ... Jacot, J. G. (2017). Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells. PLoS One, 12(5), [e0177824]. https://doi.org/10.1371/journal.pone.0177824

Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells. / Velasquez-Mao, Aaron J.; Tsao, Christopher J.M.; Monroe, Madeline N.; Legras, Xavier; Bissig-Choisat, Beatrice; Bissig, Karl Dimiter; Ruano, Rodrigo; Jacot, Jeffrey G.

In: PLoS One, Vol. 12, No. 5, e0177824, 01.05.2017.

Research output: Contribution to journalArticle

Velasquez-Mao, AJ, Tsao, CJM, Monroe, MN, Legras, X, Bissig-Choisat, B, Bissig, KD, Ruano, R & Jacot, JG 2017, 'Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells', PLoS One, vol. 12, no. 5, e0177824. https://doi.org/10.1371/journal.pone.0177824
Velasquez-Mao AJ, Tsao CJM, Monroe MN, Legras X, Bissig-Choisat B, Bissig KD et al. Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells. PLoS One. 2017 May 1;12(5). e0177824. https://doi.org/10.1371/journal.pone.0177824
Velasquez-Mao, Aaron J. ; Tsao, Christopher J.M. ; Monroe, Madeline N. ; Legras, Xavier ; Bissig-Choisat, Beatrice ; Bissig, Karl Dimiter ; Ruano, Rodrigo ; Jacot, Jeffrey G. / Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells. In: PLoS One. 2017 ; Vol. 12, No. 5.
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