Cultures of hESM human embryonic stem cells: Chromosomal aberrations and karyotype stability

Abstract

Cytogenetic analysis of karyotypes of hESM01-hESM04 human embryonic stem cells and substrains derived from these strains showed that all these strains retained normal karyotype during long-term culturing. Two substrains of embryonic stem cells with chromosome aberrations indicating clonal origin of these strains were detected. The potentialities of using analysis of chromosome variability of embryonic stem cells for evaluation of predisposition of the corresponding genotypes to the formation of chromosome abnormalities are discussed.

Chromosome rearrangements in sublines of human embryonic stem cell lines hESM01 and hESM03

“…Chromosomal rearra ngements in hESM0309 cell line Using DAPI staining, it was shown previously that in all cells of hESM0309 cell line chromosomes 4 and 9 were rearranged [5]. The patterns revealed by the staining allowed to assume that chromosome 4 contained a partial deletion and chromosome 9 a partial duplication.…”

“…Karyotyping of hESM01r18 and hESM0309 cell lines demonstrated that chromosomes r(18) and dup (9), respectively, were present in all cells from given cell lines [5]. hESM01r18 and hESM0309 cells showed a higher proliferation rate and more robust cultivation conditions compared with their parental cell lines.…”

“…(ii) Another abnormal chromosome found in hESM0309 cells was previously described as dup(9)(pter→q3::q12→qter) by DAPI banding [5]. C-banding revealed two C-positive regions confi rming the location of one of the breakpoints in a C-positive region 9q12 (Fig.…”

“…Cell lines: Sublines hESM01r18 (46,ХХ,-18,+mar) and hESM0309 (46,ХХ,del(4),dup(9)) were derived from hESM01 and hESM03 parental cell lines, respectively, as described previously [5]. The used cell lines were cultivated on 35mm petri dishes (Costar) covered with gelatine in KO-DMEM (Invitrogen), with 20% serum replacement (Invitrogen), 0.1 mM β-mercaptoethanol (Sigma), 1 mM glutamine (Gibco), 1 mM non-essential amino acids (Gibco), 4 ng/ml basic fi broblast growth factor (bFGF) (Chemicon) and antibiotics in 6.5% СО 2 on mitotically inactivated mouse embryonic fi broblasts (MEF) feeder cells.…”

“…Chromosome rearrangements in human stem cell lines were described as 46,ХХ,-18,+?r(18) [5]. The ring organization of the marker chromosome was confi rmed by the telomere specifi c probe (TTAGGG)n, which labeled termini of all normal chromosomes but gave no signal on the derivative chromosome (Fig.…”