| Chromosomal spread preparation of human embryonic stem cells for karyotyping. | |
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MedLine Citation:
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PMID: 19734841 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Priscila B Campos; Rafaela C Sartore; Stacie N Abdalla; Stevens K Rehen |
Publication Detail:
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Type: Video-Audio Media Date: 2009-09-04 |
Journal Detail:
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Title: Journal of visualized experiments : JoVE Volume: - ISSN: 1940-087X ISO Abbreviation: J Vis Exp Publication Date: 2009 |
Date Detail:
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Created Date: 2009-09-07 Completed Date: 2009-12-28 Revised Date: 2011-09-13 |
Medline Journal Info:
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Nlm Unique ID: 101313252 Medline TA: J Vis Exp Country: United States |
Other Details:
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Languages: eng Pagination: - Citation Subset: IM |
Affiliation:
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Institute of Biomedical Sciences, Federal University of Rio De Janeiro-UFRJ. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Cells, Cultured Chromosome Aberrations Chromosomes, Human* Embryonic Stem Cells / cytology, ultrastructure* Humans Karyotyping / methods* |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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