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Updated: Apr 11, 2026
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Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping
Published on: September 4, 2009
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Genomic instability of human embryonic stem cell lines using different passaging culture methods
Lucie Tosca1, Olivier Feraud2, Aurélie Magniez2
1AP-HP, Histologie-Embryologie-Cytogénétique, Hôpitaux Universitaires Paris Sud, Clamart, F-92141 France ; Université Paris Sud, Le Kremlin-Bicêtre, F-94275 France ; Esteam Paris Sud INSERM UMR-S 935, Villejuif, F-94801 France.
Molecular Cytogenetics
|June 9, 2015
Summary
Human embryonic stem cell genomic instability is similar with mechanical or enzymatic dissociation. Observed genomic variations do not impact malignancy development, regardless of dissociation method.
Area of Science:
- Stem cell biology
- Genomics
- Genomic instability
Background:
- Human embryonic stem cells (hESCs) can develop genomic instability.
- Culture duration and dissociation methods influence hESC genomic stability.
- Investigating dissociation impact on hESC genomic instability is crucial.
Purpose of the Study:
- To assess the effect of mechanical versus enzymatic (collagenase IV) dissociation on hESC genomic instability.
- To compare genomic variations in H1 and H9 cell lines at early and late passages.
Main Methods:
- Cultured H1 and H9 hESCs using mechanical/manual or enzymatic/collagenase IV dissociation.
- Evaluated genomic instability using oligonucleotide-based array-comparative genomic hybridization (105K, 50Kb resolution).
- Assessed variations at early (
Main Results:
- DNA variations primarily in subtelomeric/pericentromeric regions (<100 Kb).
- Nine recurrent genomic variations identified, including duplication 20q11.21.
- No significant differences in variation number/size, gain/loss frequencies, or gene content between dissociation methods.
Conclusions:
- Mechanical and collagenase IV enzymatic dissociation yield similar hESC genomic instability.
- Acquired genomic variations did not lead to malignancy.
- Dissociation method does not influence the development of malignancy in hESCs.
