title: Differentiation of newborn mouse skin derived stem cells into germ-like cells in vitro.
abstract: Studying germ cell formation and differentiation has traditionally been very difficult due to low cell numbers and their location deep within developing embryos. The availability of a "closed" in vitro based system could prove invaluable for our understanding of gametogenesis. The formation of oocyte-like cells (OLCs) from somatic stem cells, isolated from newborn mouse skin, has been demonstrated and can be visualized in this video protocol. The resulting OLCs express various markers consistent with oocytes such as Oct4 , Vasa , Bmp15, and Scp3. However, they remain unable to undergo maturation or fertilization due to a failure to complete meiosis. This protocol will provide a system that is useful for studying the early stage formation and differentiation of germ cells into more mature gametes. During early differentiation the number of cells expressing Oct4 (potential germ-like cells) reaches ~5%, however currently the formation of OLCs remains relatively inefficient. The protocol is relatively straight forward though special care should be taken to ensure the starting cell population is healthy and at an early passage.
authors.affiliation: Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, The University of Western Ontario, Canada. Paul.Dyce@schulich.uwo.ca
authors.forename: Paul William
authors.initials: PW
authors.lastname: Dyce
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000502:physiology; D003584:Cytological Techniques / Q000379:methods*; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D012867:Skin / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse
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