title: Isolation and culture of rabbit primordial germ cells.
abstract: Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they can also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin on inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells.
authors.affiliation: Graduate School of Biology-Oriented Science and Technology, Kinki University, Wakayama, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Kakegawa
authors.forename: Takeshi
authors.initials: T
authors.lastname: Teramura
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Takehara
authors.forename: Masayuki
authors.initials: M
authors.lastname: Anzai
authors.forename: Tasuku
authors.initials: T
authors.lastname: Mitani
authors.forename: Kazuya
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Saeki
authors.forename: Norimasa
authors.initials: N
authors.lastname: Sagawa
authors.forename: Kanji
authors.initials: K
authors.lastname: Fukuda
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Hosoi
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D002469:Cell Separation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005854:Germ Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008297:Male; D051379:Mice; D011247:Pregnancy; D011817:Rabbits / Q000196:embryology*; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism
chemical_list: D015415:Biomarkers; D036341:Intercellular Signaling Peptides and Proteins