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Updated: Jul 7, 2026
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Enrichment and Purging of Human Embryonic Stem Cells by Detection of Cell Surface Antigens Using the Monoclonal Antibodies TG30 and GCTM-2
Published on: December 6, 2013
Human embryonic stem cells passaged using enzymatic methods retain a normal karyotype and express CD30
Alison Thomson1, Davina Wojtacha, Zoë Hewitt
1Division of Gene Function and Development, Roslin Institute, Roslin, Midlothian, Scotland. Alison.Thomson@bbsrc.ac.uk
Cloning and Stem Cells
|February 5, 2008
Summary
Single cell dissociation methods impact human embryonic stem cell (hESC) karyotype stability. Trypsin-EGTA (TEG) dissociation maintained normal karyotypes, while EDTA caused rearrangements, highlighting dissociation
Area of Science:
- Stem cell biology
- Genetics
- Cell culture
Background:
- Human embryonic stem cells (hESCs) are prone to chromosomal abnormalities after single cell dissociation.
- Current dissociation methods may compromise hESC genomic integrity.
Purpose of the Study:
- To compare dissociation methods for maintaining hESC karyotype stability.
- To assess the impact of single cell dissociation on hESC genomic and phenotypic characteristics.
Main Methods:
- Compared collagenase, EDTA, and trypsin-EGTA (TEG) dissociation methods over 10 passages.
- Assessed karyotype, telomere length, pluripotency, and CD30 expression.
- Utilized competitive genomic hybridization for subkaryotypic analysis.
Main Results:
- TEG dissociation maintained normal karyotypes; EDTA caused isochromosome p7 in replicates.
- All methods retained telomere length and pluripotency.
- Subkaryotypic deletions/amplifications accumulated over time in all cultures.
- CD30 expression was upregulated on karyotypically abnormal hESCs.
Conclusions:
- Single cell dissociation using TEG can preserve karyotypically normal hESCs.
- Existing culture conditions remain suboptimal for long-term genomic stability.
- CD30 expression may serve as a marker for karyotypic abnormalities in hESCs.
