Molecular Biology and Genetics

Biotechnology Industry

Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 10 No. 2, Issue of January 15, 2007
© 2007 by Pontificia Universidad Católica de Valparaíso -- Chile Received May 11, 2006 / Accepted October 10, 2006
DOI: 10.2225/vol10-issue2-fulltext-2

Single embryoid body formation in a multi-well plate

Uthayashanker R. Ezekiel*
GeneProTech, Inc.
9103 Lawn Avenue
St. Louis, MO 63144
Tel: 314 977 8476
Fax: 314 962 3749

Mariappan Muthuchamy
Department of Systems Biology and Translational Medicine
Cardiovascular Research Institute
College of Medicine
Texas A and M University System Health Science Center
College Station, Texas 77843
Tel: 979 847 9251
Fax: 979 862 4638

Jan S. Ryerse
Department of Pathology
Saint Louis University Medical Center
Saint Louis, MO 63104
Tel: 314 977 7848
Fax: 314 977 8740

Rita M. Heuertz
Department of Clinical Laboratory Science
Doisy College of Health Sciences
Saint Louis University Medical Center
Saint Louis, MO 63104
Tel: 314 977 8610
Fax: 314 977 8503

*Corresponding author

Keywords: differentiation, EB, ES, murine embryonic stem cell.


BSA: bovine serum albumin
d.p.c.: days post coitum
EB: embryoid bodies
ES: embryonic stem
LIF: leukemia inhibitory factor
MAP2: microtubule associated protein 2
PCR: polymerase chain reaction
SEM: standard error of the mean
TBS: Tris-buffered saline

Full Text

Embryoid bodies (EB) formed from murine embryonic stem (ES) cells recapitulate many aspects of a developing embryo. Of specific importance, synchronous differentiation of EB recapitulates organ-specific development and is achieved in culture by formation of uniformly sized EB. The method described here demonstrates a simple and cost-effective way of generating EB from murine ES cells. Single EB are formed in a multi-well plate format and large numbers of EB are generated using a 96-well multi-well plate. Uniform single-sized EB formed in the multi-well are an ideal system for screening compounds and determining differentiation effects. Since EB contain all three germ layers, they are appropriate for studying small molecule effects on differentiation of ES such as is performed in high-throughput screening protocols.

Supported by UNESCO / MIRCEN network