Immunomagnetic isolation of primordial germ cells and the establishment of embryonic germ cell lines in the mouse.
The stage-specific embryonic antigen 1 (SSEA-1) is a cell marker of primordial germ cells (PGCs). In the present study, it is shown that isolation and purification of PGCs from 8.5-11.5 days post coitum (dpc) embryos can be achieved by a immunomagnetic cell sorting method using SSEA-1 antibody-conjugated magnetic beads, and then the sorted PGCs can be used for long-term culture under strict culture conditions to derive embryonic germ (EG) cell lines. Five independent EG cell lines with male karyotypes have been established. They show both a strong alkaline phosphatase activity and expression of the SSEA-1 antigen, and are karyotypically stable with a modal number of chromosomes in more than 80% of the cells. One of the EG cell lines from 8.5-dpc embryos produced chimeras after injections of the cells into 8-cell host embryos. These procedures could provide a useful and simple method for isolation of undifferentiated cells from a heterogeneous cell population and for establishment of embryo-derived stem cell lines.
Durcova-Hills
G
G
Laboratory of Reproductive Biotechnology, National Institute of Animal Industry, Tsukuba Japan.
Tokunaga
T
T
Kurosaka
S
S
Yamaguchi
M
M
Takahashi
S
S
Imai
H
H
D000818:Animals; D015415:Biomarkers; D002460:Cell Line; D002678:Chimera / Q000502:physiology*; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D005854:Germ Cells / Q000166:cytology*; D018189:Immunomagnetic Separation / Q000379:methods* / Q000662:veterinary; D016256:Lewis X Antigen / Q000276:immunology; D051379:Mice
D015415:Biomarkers; D016256:Lewis X Antigen