Establishment of embryonic stem cell lines from preimplantation mouse embryos homozygous for lethal mutations in the t-complex

Gail R. Martin, Lee M. Silver, Howard S. Fox, Alexandra L. Joyner

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Abstract

We have determined the frequency at which embryonic stem cell (ESC) lines can be established from inner cell masses (ICMs) isolated from blastocysts homozygous for lethal mutations in the mouse t-complex. Approximately one-third of the expected number, 3 29, of the ESC lines established from embryos obtained by inter-se mating of + tw18 mice are homozygous for the tw18 haplotype. These tw18 tw18 ESC lines form a variety of cell types in vitro and in vivo, including mesodermal derivatives such as cartilage and muscle. On the basis of these and data from other studies, we suggest that the normal function of the gene represented by the tw18 lethal allele is required for multiplication/survival of mesodermal precursors in the embryo rather than the specification of the mesodermal lineage, and that the lethal effects of this mutation are expressed in only the highly structured environment of the early postimplantation embryo. In studies of the lethal tw5 haplotype, we found that 2 2 ESC lines obtained are mutant homozygotes. Analysis of these data, in conjunction with the results of our earlier study (Magnuson, T., Epstein, C. J., Silver, L. M., and Martin, G. R. (1982), Nature (London) 298, 750-753), suggests that homozygosity for the genes found in the tw5 haplotype does not reduce cell viability. By contrast, 0 16 ESC lines isolated from embryos obtained from matings of + t0 mice are mutant homozygotes. Analysis of the genotypes of ICM-derived primary stem cell colonies suggests that t0 homozygous ICM cells are unable to undergo sufficient proliferation in vitro to give rise to ESC lines.

Original languageEnglish (US)
Pages (from-to)20-28
Number of pages9
JournalDevelopmental Biology
Volume121
Issue number1
DOIs
StatePublished - May 1987

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Blastocyst
Embryonic Stem Cells
Cell Line
Mutation
Embryonic Structures
Haplotypes
Homozygote
Blastocyst Inner Cell Mass
Lethal Genes
Genes
Cartilage
Cell Survival
Stem Cells
Genotype
Muscles

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

Establishment of embryonic stem cell lines from preimplantation mouse embryos homozygous for lethal mutations in the t-complex. / Martin, Gail R.; Silver, Lee M.; Fox, Howard S.; Joyner, Alexandra L.

In: Developmental Biology, Vol. 121, No. 1, 05.1987, p. 20-28.

Research output: Contribution to journalArticle

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