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    
Abstract/OtherAbstract:
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.
Authors:
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
Related Documents :
3667144 - Dynamic heterogeneity: characterization of two cell lines derived from experimental lun...
9007314 - Trisomy 13/trisomy 18 mosaicism in an infant.
2507334 - Mutant cells defective in poly(adp-ribose) synthesis due to stable alterations in enzym...
20074564 - Phenotypic characterization of mammosphere-forming cells from the human ma-11 breast ca...
20628524 - Cleavage of e-cadherin by matrix metalloproteinase-7 promotes cellular proliferation in...
21360794 - Microrna-183 family expression in hair cell development and requirement of micrornas fo...
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    
Affiliation:
Stem Cell Biology Department, National Institute for Research in Reproductive Health, Mumbai, India.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques*
Cell Differentiation
Cell Line*
Embryonic Stem Cells / cytology*,  physiology
Female
Fibroblasts / cytology
Humans
Karyotyping
Mice
Mice, SCID
Pluripotent Stem Cells / cytology,  physiology
Pregnancy
Stem Cell Transplantation
Teratoma / pathology

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


Previous Document:  Radioprotection in vitro and in vivo by minicircle plasmid carrying the human manganese superoxide d...
Next Document:  Phylogenetic modeling of heterogeneous gene-expression microarray data from cancerous specimens.