Lolait S J, Lim A T, Dahl D D, Funder J W, Toh B H
Neurosci Lett. 1983 Dec 30;43(2-3):137-42. doi: 10.1016/0304-3940(83)90177-5.
Hypothalami from 1 day neonatal rats were dissociated and cultured for 4-16 days. Using immunofluorescence and antisera against neurofilament (NF) peptides, glial fibrillary acidic protein (GFAP), galactocerebroside and fibronectin we have distinguished neurons, astrocytes, oligodendrocytes and fibroblast-like cells in culture. Astrocytes initially grew as islets of 15-30 cells which dispersed as the culture aged. These cells, together with fibronectin-reactive flat cells, formed a monolayer upon which ovoid and process-bearing cells grew after 4 days in culture. Neurofilament-positive neurons constituted 5-10% of the total cell population. In maturing cultures the number of neurons decreased and fibroblasts increased. Oligodendrocytes represented less than 1% of total cell population. These studies emphasize the necessity of using the complementary techniques of morphology and immunocytochemistry for the characterization of hypothalamic neural cells in vitro.
取1日龄新生大鼠的下丘脑进行解离,并培养4 - 16天。利用免疫荧光法以及针对神经丝(NF)肽、胶质纤维酸性蛋白(GFAP)、半乳糖脑苷脂和纤连蛋白的抗血清,我们在培养物中区分出了神经元、星形胶质细胞、少突胶质细胞和成纤维样细胞。星形胶质细胞最初以15 - 30个细胞的小岛形式生长,随着培养时间延长而分散。这些细胞与纤连蛋白反应性扁平细胞一起形成单层,培养4天后,卵形和有突起的细胞在其上生长。神经丝阳性神经元占细胞总数的5 - 10%。在成熟培养物中,神经元数量减少而成纤维细胞数量增加。少突胶质细胞占细胞总数不到1%。这些研究强调了使用形态学和免疫细胞化学互补技术来表征体外下丘脑神经细胞的必要性。