Human cardiomyocytes prior to birth by integration-free reprogramming of amniotic fluid cells

Guihua Jiang, Todd J. Herron, Julie Di Bernardo, Kendal A. Walker, K. Sue O’Shea, Shaun Kunisaki

Research output: Contribution to journalReview article

Abstract

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 β-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.

Original languageEnglish (US)
Pages (from-to)1595-1606
Number of pages12
JournalStem Cells Translational Medicine
Volume5
Issue number12
DOIs
StatePublished - Dec 1 2016
Externally publishedYes

Fingerprint

Amniotic Fluid
Cardiac Myocytes
Parturition
Transgenes
Sendai virus
Fibroblasts
Induced Pluripotent Stem Cells
Adrenergic Agonists
Skin
Regenerative Medicine
Amniocentesis
Research Ethics Committees
Karyotype
Heart Diseases
Up-Regulation
Stem Cells
Calcium
Gene Expression
Pregnancy
Population

Keywords

  • Amniotic fluid
  • Cardiac
  • Fetal stem cells
  • Induced pluripotent stem cells
  • Pluripotent stem cells

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Human cardiomyocytes prior to birth by integration-free reprogramming of amniotic fluid cells. / Jiang, Guihua; Herron, Todd J.; Di Bernardo, Julie; Walker, Kendal A.; O’Shea, K. Sue; Kunisaki, Shaun.

In: Stem Cells Translational Medicine, Vol. 5, No. 12, 01.12.2016, p. 1595-1606.

Research output: Contribution to journalReview article

Jiang, Guihua ; Herron, Todd J. ; Di Bernardo, Julie ; Walker, Kendal A. ; O’Shea, K. Sue ; Kunisaki, Shaun. / Human cardiomyocytes prior to birth by integration-free reprogramming of amniotic fluid cells. In: Stem Cells Translational Medicine. 2016 ; Vol. 5, No. 12. pp. 1595-1606.
@article{3c11bf97e8e444faa7bcdf4addac02af,
title = "Human cardiomyocytes prior to birth by integration-free reprogramming of amniotic fluid cells",
abstract = "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 β-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.",
keywords = "Amniotic fluid, Cardiac, Fetal stem cells, Induced pluripotent stem cells, Pluripotent stem cells",
author = "Guihua Jiang and Herron, {Todd J.} and {Di Bernardo}, Julie and Walker, {Kendal A.} and O’Shea, {K. Sue} and Shaun Kunisaki",
year = "2016",
month = "12",
day = "1",
doi = "10.5966/sctm.2016-0016",
language = "English (US)",
volume = "5",
pages = "1595--1606",
journal = "Stem cells translational medicine",
issn = "2157-6564",
publisher = "AlphaMed Press",
number = "12",

}

TY - JOUR

T1 - Human cardiomyocytes prior to birth by integration-free reprogramming of amniotic fluid cells

AU - Jiang, Guihua

AU - Herron, Todd J.

AU - Di Bernardo, Julie

AU - Walker, Kendal A.

AU - O’Shea, K. Sue

AU - Kunisaki, Shaun

PY - 2016/12/1

Y1 - 2016/12/1

N2 - 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 β-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.

AB - 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 β-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.

KW - Amniotic fluid

KW - Cardiac

KW - Fetal stem cells

KW - Induced pluripotent stem cells

KW - Pluripotent stem cells

UR - http://www.scopus.com/inward/record.url?scp=84995872843&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84995872843&partnerID=8YFLogxK

U2 - 10.5966/sctm.2016-0016

DO - 10.5966/sctm.2016-0016

M3 - Review article

VL - 5

SP - 1595

EP - 1606

JO - Stem cells translational medicine

JF - Stem cells translational medicine

SN - 2157-6564

IS - 12

ER -