Transgenic Research

, Volume 6, Issue 5, pp 321–328

The chromosome make-up of mouse embryonic stem cells is predictive of somatic and germ cell chimaerism

  • LETIZIA Longo
  • ANNE Bygrave
  • FRANK G. Grosveld
  • PIER PAOLO Pandolfi
Article

Abstract

Mouse pluripotent embryonic stem (ES) cells, once reintroduced into a mouse blastocyst, can contribute to the formation of all tissues, including the germline, of an organism referred to as a chimaeric. However, the reasons why this contribution often appears erratic are poorly understood. We have tested the notion that the chromosome make-up may be important in contributing both to somatic cell chimaerism and to germ line transmission. We found that the percentage of chimaerism of ES cell-embryo chimaeras, the absolute number of chimaeras and the ratio of chimaeras to total pups born all correlate closely with the percentage of euploid metaphases in the ES cell clones injected into the murine blastocyst. The majority of the ES cell clones that we tested, which were obtained from different gene targeting knockout experiments and harboured 50 to 100% euploid metaphases, did transmit to the germline; in contrast, none of the ES cell clones with more than 50% of chromosomally abnormal metaphases transmitted to the germline. Euploid ES cell clones cultured in vitro for more than 20 passages rapidly became severely aneuploid, and again this correlated closely with the percentage of chimaerism and with the number of ES cell--embryo chimaeras obtained per number of blastocysts injected. At the same time, the ability of these clones to contribute to the germline was lost when the proportion of euploid cells dropped below 50%. This study suggests that aneuploidy, rather than ‘loss of totipotency’, in ES cells, is the major cause of failure in obtaining contributions to all tissues of the adult chimaera, including the germline. Because euploidy is predictive of germline transmission, karyotype analysis is crucial and time/cost saving in any gene-targeting experiment

karyotype mouse ES cells 

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Copyright information

© Chapman and Hall 1997

Authors and Affiliations

  • LETIZIA Longo
    • 1
  • ANNE Bygrave
    • 3
  • FRANK G. Grosveld
    • 4
  • PIER PAOLO Pandolfi
    • 2
  1. 1.Department of HaematologyRoyal Postgraduate Medical SchoolLondonUK
  2. 2.Department of Human Genetics, Molecular and Cell Biology ProgramsMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  3. 3.National Institute for Medical ResearchLondonUK
  4. 4.Department of Cell Biology and Genetics, Faculty of MedicineErasmus UniversityRotterdamNetherlands

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