Immunomagnetic isolation of primordial germ cells and the establishment of embryonic germ cell lines in the mouse

• Published in: Cloning Vol. 1; no. 4; p. 217
• Main Authors: Durcova-Hills, G, Tokunaga, T, Kurosaka, S, Yamaguchi, M, Takahashi, S, Imai, H
• Format: Journal Article
• Language: English
• Published: United States 1999
• Subjects: Animals; Biomarkers; Cell Line; Chimera - physiology; Embryo, Mammalian - cytology; Embryo, Mammalian - physiology; Germ Cells - cytology; Immunomagnetic Separation - methods; Immunomagnetic Separation - veterinary; Lewis X Antigen - immunology; Mice
• ISSN: 1520-4553
• DOI: 10.1089/15204559950019852

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.