About this item:

184 Views | 103 Downloads

Author Notes:

Corresponding author: Young-Sup Yoon, MD, PhD, Division of Cardiology, Department of Medicine, Emory University School of Medicine, 1639 Pierce Drive, WMB 3009, Atlanta, GA 30322; Telephone: (404) 727-8176; Fax: (404) 727-3988; yyoon5@emory.edu

Subjects:

Research Funding:

This work was supported in part by NIH grant DP3DK094346, HHSN268201000043C, NSF-EBICS grant, CBET-0939511 and ACTSI pilot grant (PHS Grant UL1 RR025008 from the CTSA program, NIH, NCRR).

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Cardiac & Cardiovascular Systems
  • Cardiovascular System & Cardiology
  • Human embryonic stem cells
  • Human induced pluripotent stem cells
  • Cardiomyocytes
  • Directed differentiation
  • HUMAN SOMATIC-CELLS
  • FUNCTIONAL CARDIOMYOCYTES
  • CARDIAC DIFFERENTIATION
  • DEFINED FACTORS
  • BONE-MARROW
  • IN-VITRO
  • ENDODERM
  • MOUSE
  • HEART
  • LINES

Development of a novel two-dimensional directed differentiation system for generation of cardiomyocytes from human pluripotent stem cells

Journal Title:

International Journal of Cardiology

Volume:

Volume 168, Number 1

Publisher:

, Pages 41-52

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Background: Human pluripotent stem cells (hPSCs) hold great promise for treating ischemic heart disease. However, current protocols for differentiating hPSCs either result in low yields or require expensive cytokines. Methods: Here we developed a novel two dimensional (2D) stepwise differentiation system that generates a high yield of cardiomyocytes (CMs) from hPSCs without using special cytokines. Initially, undifferentiated hPSCs were transferred onto Matrigel-coated plates without forming embryoid bodies (EBs) for a few days and were cultured in bFGF-depleted human embryonic stem cells (hESCs) medium. When linear cell aggregation appeared in the margins of the hPSC colonies, the medium was changed to DMEM supplemented with 10% fetal bovine serum (FBS). Thereafter when cell clusters became visible, the medium was changed to DMEM with 20% FBS. Results and Conclusions: At about two weeks of culture, contracting clusters began to appear and the number of contracting clusters continuously increased, reaching approximately 70% of all clusters. These clusters were dissociated by two-step enzyme treatment to monolayered CMs, of which ~90% showed CM phenotypes confirmed by an α –myosin heavy chain reporter system. Electrophysiologic studies demonstrated that the hPSC-derived CMs showed three major CM action potential types with 61 to 78% having a ventricular-CM phenotype. This differentiation system showed a clear spatiotemporal role of the surrounding endodermal cells for differentiation of mesodermal cell clusters into CMs. In conclusion, this system provides a novel platform to generate CMs from hPSCs at high yield without using cytokines and to study the development of hPSCs into CMs.

Copyright information:

© 2012 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommerical-NoDerivs 3.0 Unported License (http://creativecommons.org/licenses/by-nc-nd/3.0/).

Creative Commons License

Export to EndNote