Hendriksen P J, Drews U, Frankenhuis M T, Veerhuis R, Hengst S M, Wagner U, Braun S, Booman P
DLO-Institute for Animal Science and Health (ID-DLO), Research Branch Zeist, The Netherlands.
Anat Embryol (Berl). 1994 Apr;189(4):317-25. doi: 10.1007/BF00190588.
In vitro cultures of intact chick gonads (organ cultures) and reaggregation cultures of dispersed gonad cells (roller cultures) were made. Gonads or gonad cells from 7-day-old chick embryos, at the stage when sex-specific differentiation begins, were cultured in the presence of presumed H-Y antigen-containing supernatants, or co-cultured in the presence of H-Y antigen-producing cell lines. The H-Y antigen-producing cells tested were of human, mouse, bovine and chicken origin. During organ culture, addition of supernatant of the human lymphoma cell line Daudi, or co-culture with Daudi cells, stimulated a clear proliferation of the germinal epithelium in male gonads, indicating feminization. A similar effect was obtained by treatment with estradiol. In reaggregation culture, the increase in nuclear size of germ cells was chosen as a parameter for feminization. A significant increase of germ cell nuclear size was observed in gonads cultured in the presence of Daudi supernatant. In both organ cultures and reaggregation cultures, other tested H-Y antigen sources and semi-purified H-Y antigen fractions did not exert significant effects on differentiation of the gonads or on the average area of the germ cell nuclei. These findings suggest that it is not H-Y antigen, but another protein produced by Daudi cells, that might be responsible for the sex-reversing effects.
制备了完整鸡性腺的体外培养物(器官培养)和分散性腺细胞的重聚集培养物(旋转培养)。使用来自7日龄鸡胚胎的性腺或性腺细胞,此时正是性别特异性分化开始的阶段,将其在含有假定H-Y抗原的上清液存在下进行培养,或者在产生H-Y抗原的细胞系存在下进行共培养。所测试的产生H-Y抗原的细胞来源于人、小鼠、牛和鸡。在器官培养期间,添加人淋巴瘤细胞系Daudi的上清液,或者与Daudi细胞共培养,刺激雄性性腺中生发上皮的明显增殖,表明出现雌性化。用雌二醇处理也获得了类似的效果。在重聚集培养中,选择生殖细胞核大小的增加作为雌性化的参数。在存在Daudi上清液的情况下培养的性腺中观察到生殖细胞核大小显著增加。在器官培养和重聚集培养中,其他测试的H-Y抗原来源和半纯化的H-Y抗原组分对性腺分化或生殖细胞核的平均面积均未产生显著影响。这些发现表明,可能是Daudi细胞产生的另一种蛋白质而非H-Y抗原导致了性别逆转效应。