Document Detail

Chromosomal spread preparation of human embryonic stem cells for karyotyping.
MedLine Citation:
PMID:  19734841     Owner:  NLM     Status:  MEDLINE    
Although human embryonic stem cells (hESC) have been shown to present a stable diploid karyotype, many studies have reported that depending on culture conditions they become prone to acquire chromosomal anomalies such as addition of whole or parts of chromosomes. Indeed, during long-term culture, karyotypic alterations are observed when enzymatic or chemical dissociation are used, while manual dissection of colonies for passaging retains a stable karyotype. Besides, changes in the environment such as the removal of feeder cells also seem to compromise the genetic integrity of hESC. Once chromosomal alterations could affect cellular physiology, the characterization of the genetic integrity of hESC in vitro is crucial considering hESC as an essential tool in embryogenesis studies and drug testing. Furthermore, for future therapeutic purposes chromosomal changes are a real concern as it is frequently associated to carcinogenesis. Here we show a simple and useful method to obtain high quality chromosome spreads for subsequent analysis of chromosome set by G-banding, FISH, SKY or CGH techniques. We recommend checking the chromosomal status routinely with intervals of 5 passages in order to monitor the appearance of translocations and aneuploidies.
Priscila B Campos; Rafaela C Sartore; Stacie N Abdalla; Stevens K Rehen
Publication Detail:
Type:  Video-Audio Media     Date:  2009-09-04
Journal Detail:
Title:  Journal of visualized experiments : JoVE     Volume:  -     ISSN:  1940-087X     ISO Abbreviation:  J Vis Exp     Publication Date:  2009  
Date Detail:
Created Date:  2009-09-07     Completed Date:  2009-12-28     Revised Date:  2011-09-13    
Medline Journal Info:
Nlm Unique ID:  101313252     Medline TA:  J Vis Exp     Country:  United States    
Other Details:
Languages:  eng     Pagination:  -     Citation Subset:  IM    
Institute of Biomedical Sciences, Federal University of Rio De Janeiro-UFRJ.
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MeSH Terms
Cells, Cultured
Chromosome Aberrations
Chromosomes, Human*
Embryonic Stem Cells / cytology,  ultrastructure*
Karyotyping / methods*

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