TY - JOUR
T1 - Trisomy eight in ES cells is a common potential problem in gene targeting and interferes with germ line transmission
AU - Liu, Xin
AU - Wu, Hong
AU - Loring, Janet
AU - Hormuzdi, Sheriar
AU - Disteche, Christine M.
AU - Bornstein, Paul
AU - Jaenisch, Rudolph
N1 - Medline is the source for the MeSH terms of this document.
PY - 1997/5/1
Y1 - 1997/5/1
N2 - The ability to contribute to the germ line is the most important experimental feature of embryonic stem (ES) cells. Using ES cells, it is possible to introduce targeted mutations into any gene and to derive the corresponding mutant mice. A common problem with this technology is that the ES cells often lack or have only a low efficiency of germ line transmission. To address this issue, we examined the relationship between the growth rate and karyotype of ES cells, and their ability to contribute to the germ line. We found that chromosomal abnormalities occurred rather frequently in ES cells. Cells having an abnormal number of chromosomes, in particular trisomy 8, were found in three independently derived ES cell lines, and this abnormality conferred a selective growth advantage on these cells. Selection of abnormal cells led to depletion and eventual loss of normal ES cells during consecutive passages. In comparison with parental ES cells, ES cells with trisomy 8 contributed rarely to the germ line. This realization allowed us to select, based upon ES cell clone morphology, those clones with the highest probability of contributing to the germ line. This insight is of practical value for any given gene targeting experiment as it permits optimization of the rate of success without having to rely on more elaborate tests such as karyotyping individual clones prior to blastocyst injection.
AB - The ability to contribute to the germ line is the most important experimental feature of embryonic stem (ES) cells. Using ES cells, it is possible to introduce targeted mutations into any gene and to derive the corresponding mutant mice. A common problem with this technology is that the ES cells often lack or have only a low efficiency of germ line transmission. To address this issue, we examined the relationship between the growth rate and karyotype of ES cells, and their ability to contribute to the germ line. We found that chromosomal abnormalities occurred rather frequently in ES cells. Cells having an abnormal number of chromosomes, in particular trisomy 8, were found in three independently derived ES cell lines, and this abnormality conferred a selective growth advantage on these cells. Selection of abnormal cells led to depletion and eventual loss of normal ES cells during consecutive passages. In comparison with parental ES cells, ES cells with trisomy 8 contributed rarely to the germ line. This realization allowed us to select, based upon ES cell clone morphology, those clones with the highest probability of contributing to the germ line. This insight is of practical value for any given gene targeting experiment as it permits optimization of the rate of success without having to rely on more elaborate tests such as karyotyping individual clones prior to blastocyst injection.
UR - http://www.scopus.com/inward/record.url?scp=0030944004&partnerID=8YFLogxK
U2 - 10.1002/(SICI)1097-0177(199705)209:1<85::AID-AJA8>3.0.CO;2-T
DO - 10.1002/(SICI)1097-0177(199705)209:1<85::AID-AJA8>3.0.CO;2-T
M3 - Article
AN - SCOPUS:0030944004
SN - 1058-8388
VL - 209
SP - 85
EP - 91
JO - Developmental Dynamics
JF - Developmental Dynamics
IS - 1
ER -