| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
EMBRYONIC STEM CELLS |
aWolfson Centre for Stem Cells, Tissue Engineering and Modelling, School of Human Development,
bInstitute of Genetics, and
cNottingham University Research and Treatment Unit, University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom
Key Words. Human embryonic stem cells • Embryoid body • Forced aggregation • Differentiation • Cardiomyocytes • Activin A Basic fibroblast growth factor
Correspondence: Chris Denning, B.Sc., Ph.D., University of Nottingham–Institute of Genetics, Queen's Medical Centre, Nottingham NG7 2UH, U.K. Telephone: 44(0)115-82-30385; Fax: 44(0)115-970-9906; e-mail: chris.denning{at}nottingham.ac.uk
Received September 22, 2006;
accepted for publication December 12, 2006.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLS EXPRESS December 21, 2006.
Although all human ESC (hESC) lines have similar morphology, express key pluripotency markers, and can differentiate toward primitive germ layers in vitro, the lineage-specific developmental potential may vary between individual lines. In the current study, four hESC lines were cultured in the same feeder-free conditions to provide a standardized platform for interline analysis. A high-throughput, forced-aggregation system involving centrifugation of defined numbers of hESCs in V-96 plates (V-96FA) was developed to examine formation, growth, and subsequent cardiomyocyte differentiation from >22,000 EBs. Homogeneity of EBs formed by V-96FA in mouse embryo fibroblast-conditioned medium was significantly improved compared with formation in mass culture (p < .02; Levene's test). V-96FA EB formation was successful in all four lines, although significant differences in EB growth were observed during the first 6 days of differentiation (p = .044 to .001; one-way analysis of variance [ANOVA]). Cardiomyocyte differentiation potential also varied; 9.5% ± 0.9%, 6.6% ± 2.4%, 5.2% ± 3.1%, and 1.6% ± 1.0% beating EBs were identified for HUES-7, NOTT2, NOTT1, and BG01, respectively (p = .008; one-way ANOVA). Formation of HUES-7 V-96FA EBs in defined medium containing activin A and basic fibroblast growth factor resulted in 23.6% ± 3.6% beating EBs, representing a 13.1-fold increase relative to mass culture (1.8% ± 0.7%), consistent with an observed 14.8-fold increase in MYH6 (
MHC) expression by real-time polymerase chain reaction. In contrast, no beating areas were derived from NOTT1-EBs and BG01-EBs formed in defined medium. Thus, the V-96FA system highlighted interline variability in EB growth and cardiomyocyte differentiation but, under the test conditions described, identified HUES-7 as a line that can respond to cardiomyogenic stimulation.
This article has been cited by other articles:
|
|
 |
|
  G. M. Hoben, E. J. Koay, and K. A. Athanasiou Fibrochondrogenesis in Two Embryonic Stem Cell Lines: Effects of Differentiation Timelines Stem Cells, February 1, 2008; 26(2): 422 - 430. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Allegrucci, Y.-Z. Wu, A. Thurston, C. N. Denning, H. Priddle, C. L. Mummery, D. Ward-van Oostwaard, P. W. Andrews, M. Stojkovic, N. Smith, et al. Restriction landmark genome scanning identifies culture-induced DNA methylation instability in the human embryonic stem cell epigenome Hum. Mol. Genet., May 15, 2007; 16(10): 1253 - 1268. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| STEM CELLS | THE ONCOLOGIST | CME | ALPHAMED PRESS JOURNALS |
