Document Detail

Derivation and characterization of two genetically unique human embryonic stem cell lines on in-house-derived human feeders.
MedLine Citation:
PMID:  18699724     Owner:  NLM     Status:  MEDLINE    
This study describes the successful derivation of two human embryonic stem (hES) cell lines using 53 frozen and 18 fresh "slow-growing" surplus embryos, obtained from collaborating in vitro fertilization clinics, on in-house-derived human feeder layers. The cell lines have been derived by whole embryo culture followed by further expansion of manually dissected inner cell mass from the surrounding trophoectodermal cells. Immunocytochemical localization of cell surface markers like SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81, staining for alkaline phosphatase and reverse transcriptase polymerase chain reaction (RT-PCR) analysis of pluripotency state markers viz. Oct-4, TERT, Nanog, Rex1, and Sox2 indicate that both cell lines possess typical features of embryonic stem cells. Both cell lines exhibit normal female karyotype after 40 passages in culture. Pluripotent nature of the cell lines was confirmed both in vitro and in vivo. Embryoid bodies, formed in suspension culture, express markers for all three lineages as indicated by RT-PCR analysis for SOX 1 (ectoderm), HAND 1 (mesoderm), AFP (endoderm), and CDX2 (trophoectoderm). Teratoma formed in vivo in severe combined immunodeficient mice revealed cells of all the three embryonic germ layers. Comparison of the STR and human leukocyte antigen profiles of these cell lines with the existing human ES cell lines indicate that they are genetically distinct. The addition of our hES cell lines contributes usefully to the globally restricted repertoire of human ES cell lines.
Neeraj Kumar; Indira Hinduja; Punam Nagvenkar; Lakshmi Pillai; Kusum Zaveri; Leena Mukadam; Jyoti Telang; Sadhana Desai; Vijay Mangoli; Ranjana Mangoli; Shreyas Padgaonkar; Gurvinder Kaur; Chander Puri; Deepa Bhartiya
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Stem cells and development     Volume:  18     ISSN:  1557-8534     ISO Abbreviation:  Stem Cells Dev.     Publication Date:  2009 Apr 
Date Detail:
Created Date:  2009-04-20     Completed Date:  2009-06-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101197107     Medline TA:  Stem Cells Dev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  435-45     Citation Subset:  IM    
Stem Cell Biology Department, National Institute for Research in Reproductive Health, Mumbai, India.
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MeSH Terms
Cell Culture Techniques*
Cell Differentiation
Cell Line*
Embryonic Stem Cells / cytology*,  physiology
Fibroblasts / cytology
Mice, SCID
Pluripotent Stem Cells / cytology,  physiology
Stem Cell Transplantation
Teratoma / pathology

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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