Tamura S
Nihon Jibiinkoka Gakkai Kaiho. 1989 Jul;92(7):1084-92. doi: 10.3950/jibiinkoka.92.1084.
Rat embryos were explanted on late 11 day of gestation and cultured for 24 hours in rotating bottles with the yolk sac opened. Rat serum was used as culture medium and culture bottles were filled with 5% CO2 + 95% O2 gas mixture as gas phase. At the time of explantation and the end of the culture period, differentiation and growth of the embryos were monitored by counting somites and measuring crown rump length. About part of the embryos, protein determinations were made to measure growth. These data were compared with values found for 12 and 13 day embryos. The results for cultured embryos showed slight retardation in their differentiation and slight depression in the growth. At the same time to study early inner ear development in vitro, 9 of the other cultured embryos were serially sectioned and observed by light microscopy. In the cultured embryos endolymphatic duct elongated and acoustico-facial ganglion enlarged remarkably. Otocysts became flattened and elongated ventrally. Vestibular and cochlear portion were identifiable in the otocysts. Nerves arose from the ganglion reached to brain centrally and wall of pharynx peripherally. There was no significant difference in inner ear development between 13 day and the cultured embryos. This culture system should prove useful for studies on early inner ear development of mammalian embryos.
将妊娠第11天晚期的大鼠胚胎取出，在卵黄囊打开的旋转瓶中培养24小时。以大鼠血清作为培养基，培养瓶中充入5%二氧化碳 + 95%氧气的混合气体作为气相。在植入时和培养期结束时，通过计数体节和测量顶臀长度来监测胚胎的分化和生长。对部分胚胎进行蛋白质测定以测量生长情况。将这些数据与12天和13天胚胎的数值进行比较。培养胚胎的结果显示其分化略有延迟，生长略有抑制。同时，为了在体外研究早期内耳发育，对另外9个培养胚胎进行连续切片并通过光学显微镜观察。在培养的胚胎中，内淋巴管延长，听面神经节明显增大。耳囊腹侧变平并拉长。在耳囊中可识别出前庭和耳蜗部分。从神经节发出的神经向中枢延伸至脑，向外周延伸至咽壁。13天胚胎和培养胚胎的内耳发育没有显著差异。这种培养系统对于研究哺乳动物胚胎的早期内耳发育应该是有用的。
