マウス胚の予定生殖巣および生殖巣原基の体外培養

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Explantation of tissues of prospective gonads and gonad primordia of mouse embroys was performed with the view of studying the morphogenic mechanism of sex differentiation of mammalian gonads. The explants taken from mouse embroys ranging in age from the 12th to the 16th day after impregnation was cultured in vitro, employing the rolling tube culture method. The fluid medium used in the present study consisted of Tyrode solution and horse serum at the ratio of 1:1 or 2:1. A gaseous medium of 98% oxygen and 2% carbon dioxide was also employed to adjust the pH of the culture fluid. Penicillin G in concentration of 2 mg/1 was added to the fluid medium for the purpose of asepsis. Cultures were placed at a slant and rolled at a speed of 1/3 r.p.m., being held at 37.5℃ in the incubator. Culture medium was not changed throughout the period of incubation. In all of the experiments the explants carried on their development for a period of 21-44 hours, irrespective of the ratio of the constitution of the fluid medium, and realized the sex differentiation which was almost similar in their histologic features to that of the gonad in vivo of the corresponding ages, though their general development was retarded in some degree. The normal development of the cultures is limited within 70 hours of incubation, and thereafter the explants showed somewhat atrophic features. It was found that the prospective gonad of the embryos at the 12th day after gestation which was histologically indifferent in regard to sex, had a capacity to differentiate sexually even in vitro. Namely, the gonad of the said developmental stage has a good capability of auto-differentiation. When the explant reached the stage of the 14-15th day in age, their Mullerian ducts atrophied in compliance with its developmental fate just in the same way as is ordinarily observed in the normal development of gonads of the mouse. It was ascertained that the experimental technique employed in the present study is as a whole a good expedient in the analysis of the gonad differentiation of the mammal, at least of mouse embryos, though the constitution of culture media leaves something to be improved. Explanation of Plates Figures of series a show the general appearance of the prospective gonads (or the gonads) and the surrounding tissues in vivo or in vitro. The magnification of all photographs is ×100. Figures of series b show the detail of the gonadal part of the figure of the corresponding number, in higher magnification, ×400. Abbreviation: bc blastem cell; bl basal layer; g gonadal tissue; gep germinal epithelium; ib interstitial blastema; ic interstitial cell; ict interstitial connective tissue; it interstitial tissue; l liver; md Mullerian duct; mes mesonephros; met metanephros; og oogonia; pep peritoneal epithelium; pgc primordial germ cell; psc primary sex cord; pst primodial seminiferous tubule; sc sertoli's cell; sg spermatogonia; sga abnomal spermatogonia; ssc secondary sex cord; ta tunica albuginea; tv tunica vasculosa; wd wolffian duct. Plate II 1. Prospective gonad and gonad-forming area of the embryo at the 12th day after impregnation (designated in the following as 12-day embryo). 2. Cultured gonad-forming area derived from 12-day embryo, cultured for 21 hours. 3. Gonad of primary testicular differentiation, from 14-day embryo. 4. Gonad of primordial ovarial differentiation, from 14-day embryo. Plate III 5. Cultured gonad differentiating into primary testis, derived from 12-day embryo, cultured for 44 hours. 6. Cultured gonad showing ovarial differentiation, derived from 12-day embryo cultured for 44 hours. 7. Cultured gonad showing testicular development, from 13-day embryo cultured for 21 hours. 8. Uncultured gonad of testicular development, from 15-day embryo. Plate IV 9. Cultured gonad presenting the testicular differentiation, from 13-day embryo, cultured for 44 hours. 10. Uncultured gonad in testicular development, from 16-day embryo. 11. Cultured testis, from 1
社団法人日本動物学会の論文
1960-09-15

マウス胚の予定生殖巣および生殖巣原基の体外培養

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