title: Isolation and characterization of embryonic stem cell-like cells from in vitro produced goat (Capra hircus) embryos.
abstract: The aim of the present study was to isolate and characterize goat embryonic stem cell-like cells from in vitro produced goat embryos. Inner cell mass (ICM) cells were isolated either mechanically or by enzymatic digestion from 150 blastocysts and 35 hatched blastocysts whereas 100 morulae were used for blastomeres isolation mechanically. The ICM derived cells or blastomeres were cultured on a feeder layer. The primary colony formation was significantly higher (P < 0.01) for hatched blastocysts (77.14%) than early/expanded blastocysts (54%) or morula (14%). When ICMs were isolated mechanically the primary colony formation for hatched blastocysts (90%) as well as blastocysts (66%) were significantly more than when ICMs were isolated by enzymatic digestion (60% and 30%, respectively). The colonies were disaggregated either mechanically or by enzymatic digestion for further subculture. When mechanical method was followed, the colonies remained undifferentiated up to 15 passages and three ES cell-like cell lines were produced (gES-1, gES-2, and gES-3). However, enzymatic disaggregation resulted in differentiation. The undifferentiated cells showed stem cell like morphological features, normal karyotype, and expressed stem cell specific surface markers like alkaline phosphatase, TRA-1-61, TRA-1-81, and intracellular markers Oct4, Sox2, and Nanog. Following prolonged culture of the ES cell-like cells were differentiated into several types of cells including neuron like and epithelium-like cells. In conclusion, goat embryonic stem cell-like cells can be isolated from in vitro produced goat embryos and can be maintained for long periods in culture.
authors: Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors: Arun
authors: A
authors: Kumar De
authors: Dhruba
authors: D
authors: Malakar
authors: Yogesh S
authors: YS
authors: Akshey
authors: Manoj Kumar
authors: MK
authors: Jena
authors: Rahul
authors: R
authors: Dutta
mesh_terms: D000818:Animals; D053624:Blastocyst Inner Cell Mass / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation; D002478:Cells, Cultured; D004587:Electrophoresis, Agar Gel; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D058732:Embryoid Bodies; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology; D006041:Goats / Q000196:embryology*; D059785:Karyotype; D008853:Microscopy; D009028:Morula / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction
chemical_list: D050814:Octamer Transcription Factor-3