title: Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells.
abstract: This chapter describes standard techniques to (1) maintain mouse embryonic stem cell culture, (2) deliver transgenes into mouse embryonic stem cells mediated by electroporation, nucleofection, lipofection, and retro/lentiviruses, and (3) assess the pluripotency of mouse embryonic stem cells. The last part of this chapter presents induction of random cell differentiation followed by the alkaline phosphatase and embryoid body formation assays, immunofluorescence microscopy, and the teratoma formation assay.
authors: Department of Biological Sciences, Chemical and Biomolecular Engineering, University of Notre Dame, 49 Galvin Life Sciences, Notre Dame, IN, 46556, USA. Tetsuya.Tanaka.9@nd.edu.
authors: Tetsuya S
authors: TS
authors: Tanaka
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D018274:Electroporation / Q000379:methods; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D018014:Gene Transfer Techniques; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D019076:Transgenes
chemical_list: D004247:DNA
keywords: Alkaline phosphatase; Electroporation; Embryoid body; Immunofluorescence microscopy; Lentivirus; Lipofection; Mouse embryonic stem cells (mESCs); Nucleofection; Pluripotency; Retrovirus; Self-renewal; Teratoma