Development of a reproducible in vitro method to grow and study differentiation of mouse embryonic stem cells in the presence of the vitamin A derivative, retinoic acid

Christopher Wesley Norwood, Tennessee State University

Abstract

Mouse embryonic stem cells (mESC) are pluripotent giving them the ability to differentiate into various specialized cell types. mESC can be grown in medium containing fetal bovine serum (FBS) and Leukemia Inhibitory Factor (LIF) on a mouse embryonic fibroblast (MEF) feeder layer. These growth conditions confound studies of growth factor effects because there are undefined factors in FBS and MEFs. Austin Smith's laboratory has developed a serum-free medium containing three defined growth factors, LIF, Bone Morphogenetic Factor (BMP4) and Glycogen Synthase Kinase 3β inhibitor (GSK3βi) that allows growth of mESCs on gelatinized culture dishes. We confirmed that C57/BL6 mESC do maintain pluripotency when grown using Smith's media, and extended this work by developing a method for growing the cells on non-gelatinized slides that permitted better growth. Development of this reproducible method was then used to begin studying the effects of retinoic acid (RA) on neuron differentiation. ^

Subject Area

Biology, Neuroscience|Biology, Cell|Health Sciences, Human Development

Recommended Citation

Christopher Wesley Norwood, "Development of a reproducible in vitro method to grow and study differentiation of mouse embryonic stem cells in the presence of the vitamin A derivative, retinoic acid" (2011). ETD Collection for Tennessee State University. Paper AAI1492284.
http://digitalscholarship.tnstate.edu/dissertations/AAI1492284

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