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Production of Cloned and Transgenic Embryos Using Buffalo (Bubalus bubalis) Embryonic Stem Cell-Like Cells Isolated from In Vitro Fertilized and Cloned Blastocysts.
MedLine Citation:
PMID:  21548826     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Abstract Here, we report the isolation and characterization of embryonic stem (ES) cell-like cells from cloned blastocysts, generated using fibroblasts derived from an adult buffalo (BAF). These nuclear transfer embryonic stem cell-like cells (NT-ES) grew in well-defined and dome-shaped colonies. The expression pattern of pluripotency marker genes was similar in both NT-ES and in vitro fertilization (IVF) embryo-derived embryonic stem cell-like cells (F-ES). Upon spontaneous differentiation via embryoid body formation, cells of different morphology were observed, among which predominant were endodermal-like and epithelial-like cell types. The ES cell-like cells could be passaged only mechanically and did not form colonies when plated as single cell suspension at different concentrations. When F-ES cell-like, NT-ES cell-like, and BAF cells of same genotype were used for hand-made cloning (HMC), no significant difference (p > 0.05) was observed in cleavage and blastocyst rate. Following transfer of HMC embryos to synchronized recipients, pregnancies were established only with F-ES cell-like and BAF cell-derived embryos, and one live calf was born from F-ES cell-like cells. Further, when transfected NT-ES cell-like cells and BAF were used for HMC, no significant difference (p > 0.05) was observed between cleavage and blastocyst rate. In conclusion, here we report for the first time the derivation of ES cell-like cells from an adult buffalo, and its genetic modification. We also report the birth of a live cloned calf from buffalo ES cell-like cells.
Authors:
Aman George; Ruchi Sharma; Karn P Singh; Sudeepta K Panda; Suresh K Singla; Prabhat Palta; Radhaysham Manik; Manmohan S Chauhan
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-5-6
Journal Detail:
Title:  Cellular reprogramming     Volume:  -     ISSN:  2152-4998     ISO Abbreviation:  -     Publication Date:  2011 May 
Date Detail:
Created Date:  2011-5-9     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101528176     Medline TA:  Cell Reprogram     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Embryo Biotechnology Lab, Animal Biotechnology Centre , National Dairy Research Institute, Karnal-132001, India .
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