title: In vitro culture of embryonic disc cells from porcine blastocysts.
abstract: The aim of the present study was to define the conditions of preparation and in vitro culture of embryonic discs allowing proliferation of ES-like cells. G5-6 porcine blastocysts (G0 = day of AI) were cultured in toto; in G10-11 blastocysts, trophectoderm and primitive endoderm were microsurgically removed from embryonic discs (ED) which were cultured either on plastic or on a feeder layer. Feeder cells were foetal G30 porcine fibroblasts which had been previously irradiated. Culture medium was DMEM supplemented with 0.1 mM beta-mercaptoethanol, 5% foetal calf serum, 5% Ultroser G and 10(3) IU LIF; cultures were performed at 38 degrees C. Colonies were reseeded weekly. Few embryonic discs from G5-6 and no elongating blastocysts gave rise to ES-like cells. At least 50% G10-11 ED attached and developed multilayered colonies (100 cells) of small ovoid ES-like cells. Colonies from 4 sows were maintained in culture for at least 8 wk. Addition of PDGF, insulin or both, induced a transitory stimulation of growth in G6 or G10-11 ED; TGF beta did not modify growth of G6 ICM. Uterine G10-11 flushing medium or retinol induced differentiation of ES-like cells. These cells introduced in nude mice induced teratoma.
authors.affiliation: URA INRA-CNRS, 1291 Physiologie de la Reproduction des Mammifères Domestiques, Nouzilly, France.
authors.forename: M T
authors.initials: MT
authors.lastname: Hochereau-de Reviers
authors.forename: C
authors.initials: C
authors.lastname: Perreau
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods; D005347:Fibroblasts / Q000528:radiation effects; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D008819:Mice, Nude; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D013552:Swine / Q000196:embryology*; D013724:Teratoma / Q000209:etiology / Q000473:pathology; D016211:Transforming Growth Factor alpha / Q000494:pharmacology; D014801:Vitamin A / Q000494:pharmacology
chemical_list: D003470:Culture Media; D007328:Insulin; D010982:Platelet-Derived Growth Factor; D016211:Transforming Growth Factor alpha; D014801:Vitamin A