Chromosomal integrity maintained in five human embryonic stem cell lines after prolonged in vitro culture

• There have been recent reports of human embryonic stem cell (hESC) lines developing chromosomal aberrations after long-term culture, indicating an unstable genomic status due to the in vitro milieu. This raises concern, since it would limit their use in therapeutics. In this study the chromosomal status of five well-characterized hESC lines, SA002, SA002.5, AS034.1.1, SA121 and SA461, was monitored during long-term in vitro culture. The criteria of defined hESCs were met by all of the five hESC lines (four diploid and one trisomic for chromosome 13). The genomes were screened for chromosomal aberrations and rearrangements using comparative genomic hybridization (CGH), interphase fluorescence in situ hybridization (FISH) and traditional karyotyping on several occasions while in culture. The genomic integrity was shown to be maintained after repeated freeze-thaw procedures and continuous culture in vitro for up to 22 months (148 passages). We discuss the most common de novo chromosomal aberrations reported in hESCs, as well as their possible origin.

Subject headings

NATURVETENSKAP  -- Matematik (hsv//swe)

NATURAL SCIENCES  -- Mathematics (hsv//eng)

Keyword

*Cell Culture Techniques

Cell Line

Cell Proliferation

*Chromosomal Instability

Chromosome Aberrations

Chromosomes

Human

Embryo/*cytology

Female

Humans

In Situ Hybridization

Fluorescence

Karyotyping

Nucleic Acid Hybridization

Retrospective Studies

Stem Cells/*metabolism

Publication and Content Type

ref (subject category)

art (subject category)