Detection of recurrent chromosome 12 aneuploidy in human induced pluripotent stem cells using FISH with custom centromeric DNA-probes

Background. Approximately 20% of induced pluripotent stem cell (iPSC) lines spontaneously acquire chromosomal abnormalities, which hinders their use in research and medical applications. One rational approach for routine monitoring of genetic stability in iPSCs is the targeted detection of recurrent karyotype anomalies, 20-60% of which may involve trisomy of chromosome 12. Trisomy 12 leads to impaired differentiation capacity and replication dynamics and is associated with the rapid displacement of the euploid clone.

Aim. To develop a protocol for the production of fluorescent DNA probes for the detection of chromosome 12 aneuploidy.

Results. A two-stage protocol for obtaining DNA probes based on PCR, using genomic DNA as the template, has been developed, followed by direct click-labeling with a fluorochrome. Hybridization conditions have been optimized for iPSC lines with known euploid and aneuploid karyotypes, confirming the effectiveness and specificity of detecting chromosome 12 on metaphase spreads and in     interphase nuclei.

Conclusions. FISH using custom DNA probes enhances the availability of chromosome 12 copy number analysis in laboratories, facilitating the timely detection of functionally significant genetic abnormalities in cell lines.

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