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vol.19 issue2TEACHING HUMAN ANATOMY: EXPERIENCE AND CHALLENGE IN A MEDICAL SCHOOLMORPHOLOGIC STUDY OF THE BULBOURETHRAL GLAND OF THE RABBIT (Oryctolagus cuniculus) author indexsubject indexarticles search
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Revista chilena de anatomía

Print version ISSN 0716-9868

Abstract

ROJAS, Mariana et al. CHARACTERIZATION, ISOLATION AND CULTURE OF RABBIT PRIMORDIAL GERMINAL CELLS. Rev. chil. anat. [online]. 2001, vol.19, n.2, pp.213-220. ISSN 0716-9868.  http://dx.doi.org/10.4067/S0716-98682001000200014.

Primordial germ cells (PGC) directly obtained from the embryo are not able to start pluripotent cell lines but they do so if cultured in vitro. The aims of this work are. 1) characterize by immunohistochemistry the rabbit PGC in their different "in vivo" embryonary stages; 2) identify the optimal site to obtain these cells according to embryonic age and 3) to evaluate if murine feeder cells permit proliferation and survival of PGC cultured "in vitro". Embryos were obtained from 16 White New Zealand rabbits of 7, 9, 10-14 and 16 days post coitum (dpc). A group of each litter was processed "in toto"and in frozen sections for stainning for alkaline phosphatase and two monoclonal antibodies, TEC-1 (identifies the PGC surface antigen SSEA-1), PG-2 that stains PGC perimitochondrial cytoplasm. Another group of embryos of each litter were cultured "in vitro" for 22 days using STO and MI-220 as feeder cells. In 7 days embryos, the epiblast of the embryonic disk present TEC-1 possitive and PG-2 negative cells. This mark is retained during the transitory stage in the alantoid and migration along the intestinal mesentery. When PGC colonize the gonad they become TEC-1 negative and PG-2 positive.  "In vitro"  proliferative ability of rabbit PGC is less than that observed in mice. The former cells have a reduced proliferation also "in vivo". The "in vitro" proliferative efficiency of PGC correlates well with the age of the embryo from which they derive and so does also their morphology.  In conclusion, the optimal age for best proliferation and survival "in vitro" was at 14 days, that is the period when PGC are proliferating and migrating along the intestinal mesentery "in vivo". The murine culture medium with STO and MI-220 allow proliferation and survival of rabbit PGC.

Keywords : Embryology; Primordial germ cells; Cell culture; Antibodies TEC-1, PG-2.

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