Monolayer Culture Condition for Mouse Embryonic Stem Cells Differentiation into Neural Crest Cells (Method)
Understanding the molecular mechanisms underlying craniofacial development would encourage the development of alternative in vitro embryo-toxicity tests and contribute to a decreased incidence of drug-induced congenital anomalies in the craniofacial region, such as cleft lip or cleft palate. During craniofacial development, neural crest-derived mesenchymal cells migrate to the branchial arches and contribute extensively to formation of maxillofacial structures, neural cells, smooth muscle cells, bone and cartilage cells, and odontoblasts. The neural crest (NC) is a group of cells located in the neural folds at the boundary between the neural and epidermal ectoderm. The molecular mechanisms underlying NC induction during early development remain poorly understood. We previously established a defined serum-free culture condition for mouse embryonic stem (mES) cells without feeders. Use of culture media containing undefined or undisclosed components has limited the development of applications for pluripotent cells because of our lack of knowledge of their responses to specific cues that control self-renewal, differentiation, and lineage selection. Using the defined condition in this study, we developed a protocol to promote mES cell differentiation into NC cells in an adherent monolayer culture. In the culture conditions, the effect of exogenous factors can be analyzed without confounding influences of undefined components. This protocol could be used to help clarify the mechanisms underlying craniofacial development and to assist the development of in vitro alternative developmental toxicity test for drug-induced congenital anomalies in the craniofacial region.
KeywordsNeural Crest Leukemia Inhibitory Factor Neural Crest Cell Mouse Embryonic Stem Cell Craniofacial Development
I thank Prof. Makoto Asahima and Prof. Hideyuki Okano for their valuable comments on the concept of neural cell differentiation. I thank Dr. J. Denry Sato for the valuable comments on the concept of growth factor defined serum-free culture methods and also editorial assistance.
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