Establishment of an embryonic stem cell line from 8-cell stage mouse embryos.
The aim of this study was to try establishing mouse ES cell lines from the early developmental stage. Fifty-two uncompacted 8-cell stage embryos were dissociated and single blastomeres were seeded on primary embryonic fibroblasts in DMEM/F12 completed with 10% foetal calf serum, 10% new born calf serum. 10(-4) M beta-mercaptoethanol. After approximately 5 days of culture, multiple cell clones exhibiting stem cell morphology grew out and were dissociated. One cell line was established (MSB1) and characterised. The karyotype and the G-banding revealed a male diploid cell line. MSB1 cells were injected into syngenic mice and produced teratocarcinomas. Detailed histological examination of the tumours showed a great variety of cell types including representatives of all three primary germ layers. Several nests of undifferentiated stem cells were also present. Microinjections of MSB1 cells into 52 blastocysts produced 2 chimeras, 1 male and 1 female. These results demonstrate that a highly pluripotents ES cell line can be derived from 8-cell stage mouse embryos. However, the male chimera appeared sterile. More experiments would thus be necessary to prove that the cell line obtained is capable to colonise the germ line.
Delhaise
F
F
Université Catholique de Louvain, Unité des Sciences Vétérinaires, Louvain-la-Neuve, Belgique.
Bralion
V
V
Schuurbiers
N
N
Dessy
F
F
D000818:Animals; D001284:Atrophy; D001757:Blastomeres / Q000166:cytology*; D015197:Carcinogenicity Tests; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line / Q000166:cytology / Q000637:transplantation; D002678:Chimera; D005260:Female; D007621:Karyotyping; D008297:Male; D051379:Mice; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology*; D018243:Teratocarcinoma; D013737:Testis / Q000473:pathology