Controlling embryonic stem cell growth and differentiation by automation: enhanced and more reliable differentiation for drug discovery.
Despite significant use in basic research, embryonic stem cells have just begun to be used in the drug discovery process. Barriers to the adoption of embryonic stem cells in drug discovery include the difficulty in growing cells and inconsistent differentiation to the desired cellular phenotype. Embryonic stem cell cultures require consistent and frequent handling to maintain the cells in a pluripotent state. In addition, the preferred hanging drop method of embryoid body (EB) differentiation is not amenable to high-throughput methods, and suspension cultures of EBs show a high degree of variability. Murine embryonic stem cells passaged on an automated platform maintained ≥ 90% viability and pluripotency. We also developed a method of EB formation using 384-well microplates that form a single EB per well, with excellent uniformity across EBs. This format facilitated high-throughput differentiation and enabled screens to optimize directed differentiation into a desired cell type. Using this approach, we identified conditions that enhanced cardiomyocyte differentiation sevenfold. This optimized differentiation method showed excellent consistency for such a complex biological process. This automated approach to embryonic stem cell handling and differentiation can provide the high and consistent yields of differentiated cell types required for basic research, compound screens, and toxicity studies.
Beckman Coulter, Inc., Indianapolis, IN 46268, USA. mkowalski@beckman.com
Michael P
MP
Kowalski
Amy
A
Yoder
Li
L
Liu
Laura
L
Pajak
D000818:Animals; D001331:Automation / Q000379:methods*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D055808:Drug Discovery / Q000379:methods*; D058732:Embryoid Bodies; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism