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Zhang S.,Northwest University, China | Zhang S.,Shaanxi Center for Stem Cell Engineering and Technology | Teng Y.,Arizona State University
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis | Year: 2013

Generating homozygous mutants in mammalian cells has been complicated by their diploid genome. If one allele of an autosomal gene was disrupted, the resulting heterozygous mutant is unlikely to display a phenotype due to the existence of the other allele. Although embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are excellent cellular models for analyzing developmental events or disease phenotypes in vitro, a direct analysis of recessive phenotypes has been limited by their diploidy. Recently, four independent research groups reported successful derivation of haploid mouse embryonic stem cells which provide an effective platform for studying mammalian gene function. The aim of this review is to describe the strategies for deriving haploid ESCs and compare their characteristics with diploid ESCs, and further discuss the potential application of haploid ESCs in genetic screening and homozygous mutant animal production. © 2013 Elsevier B.V.

Jia W.,Shaanxi Center for Stem Cell Engineering and Technology | Cheng D.,Shaanxi Center for Stem Cell Engineering and Technology | Chen S.,Shaanxi Center for Stem Cell Engineering and Technology | Lei L.,Shaanxi Center for Stem Cell Engineering and Technology | And 2 more authors.
Cell Biology International | Year: 2011

Spermatogonia and sperm-like cells can be derived in vitro via the addition of RA (retinoic acid) to pluripotent ES and EG cells. At present, however, these cells have not been derived from unipotent cells. Here, we have generated premeiotic Stra8-positive cells from C2C12 myoblasts following treatment with 10 mM all-trans-RA for 8 days. The differentiated C2C12 cells exhibited spherical morphology similar to spermatogonia, and they expressed gene markers of premeiosis, meiosis and postmeiosis. In addition, some of the transdifferentiated Stra8-positive cells had a tail-like phenotype. Flow cytometry results indicated that up to 20% of RA-induced C2C12 cells were Stra8-positive. Mvh (mouse vasa homologue) protein, a germ cell-specific ATP-dependent RNA helicase and Prm1 (protamine 1) were detected in transdifferentiated cells. The DNA content in induced C2C12 cells showed that Stra8-positive cells were diploid, suggesting that the myoblast transdifferentiation was in the premeiotic stage of spermatogenesis. The derivation of Stra8-positive cells from C2C12 myoblasts has important implications for studying unipotent cell differentiation. Furthermore, C2C12 myoblasts may provide a useful in vitro cell model to study signal transduction and transdifferentiation during RA treatments. © The Author(s) Journal compilation. © 2011 Portland Press Limited.

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