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Regen Med. 2009 Mar;4(2):225-37. doi: 10.2217/17460751.4.2.225.

Cardiomyocyte enrichment from human embryonic stem cell cultures by selection of ALCAM surface expression.

Author information

  • 1Lonza Walkersville, Inc., 8830 Biggs Ford Road, Walkersville, MD 21793, USA. william.rust@lonza.com

Abstract

AIMS:

The production of a homogenous population of human cardiomyocytes that can be expanded in vitro may facilitate development of replacement tissue lost as a result of cardiac disease and injury.

MATERIALS AND METHODS:

We evaluated the utility of activated leukocyte cell-adhesion molecule, CD166 (ALCAM) expression as a marker for isolating cardiomyocytes from differentiating cultures of human embryonic stem cells (hESCs). Using RT-qPCR, immunohistochemistry and DNA methylation studies, we evaluated the developmental age of hESC-derived cardiomyocytes.

RESULTS AND CONCLUSIONS:

We demonstrate that cardiomyocytes derived from hESC cultures express ALCAM and that this surface antigen can be used to select a population of differentiated cells that are enriched for cardiomyocytes. Expression of contractile proteins and ion channels, and DNA methylation patterns, suggest that ALCAM-enriched cardiomyocytes have an embryonic phenotype. Selected cardiomyocyte populations survive sorting, adhere to collagen-coated tissue culture plastic and proliferate in short-term culture. Long-term in vitro survival of cardiomyocytes was achieved by culturing cells in 3D aggregates.

PMID:
19317642
[PubMed - indexed for MEDLINE]
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