Non-clonal mosaicism in human somatic and embryonic stem cells revealed by single-cell array-based copy-number variation analysis

ABSTRACT: Although it is known that cultured human cells acquire copy number variations over time, little is known about the mutation frequency in individual cells. Here we describe that human somatic and embryonic stem cell cultures show significant fractions of cells carrying unique chromosomal abnormalities, forming a non-clonal genetic mosaic. We studied 85 human single cells by array-based comparative genomic hybridisation and found that 14-31% of hESC and 8-26% of somatic cells are chromosomally abnormal. Remarkably, only 2 cells showed full-chromosome aneuploidy, while 93% of detected abnormalities were segmental, most of them telomere-spanning. Furthermore, fluorescent in situ hybridisation confirmed this finding and revealed an increased instability of the subtelomeric regions in hESC as compared to somatic cells. We first validated the aCGH method by studying single cells with known genetic imbalances of different sizes. We analysed 2 amniocytes (Amniocyte C 01 - Amniocyte C 02) with a trisomy 18, eight amniocytes carrying a deletion of 5p33.3p15.55 and a duplication of 9q33q34.3 (Amniocyte A 01 - Amniocyte A 08), two amniocytes carrying a deletion of 5p15.33p15.1 (Amniocyte B 01 - Amniocyte B 02) and three hESC carrying a duplication of 3q26.33q27.3 (VUB07 P165BBB7 01 - VUB07 P165BBB7 03). From this, we found that we can reliably detect at the single-cell level CNVs that span seven consecutive BAC clones on the microarray.

ORGANISM(S): Homo sapiens

SUBMITTER: Kurt Jacobs 

PROVIDER: E-GEOD-51126 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress