Nishimura Shotaro, Yamahira Shungo, Chowdhury Vishwajit S, Hosaka Yoshinao Z
Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
School of Agriculture, Kyushu University, Fukuoka, Japan.
Anim Sci J. 2023 Jan-Dec;94(1):e13814. doi: 10.1111/asj.13814.
Chicken adenohypophyseal cells were cultured in plates coated with different materials, and their morphologies were examined to confirm the characteristics of chicken folliculo-stellate (FS) cells in vitro. The adenohypophyseal cells were dispersed with a collagenase/trypsin mixture in media and seeded in plates coated in either poly L-lysine (PLL), collagen, or laminin. After 7 days of culture, the cells were fixed and immunocytochemistry was performed. 5-Bromo-2'-deoxyuridine incorporation test indicated that the proliferation activity of the culture cells was different based on the coating materials, and it was higher in the collagen-coated plate than two other coating materials. Fluorescence immunocytochemistry was also performed using mixed antibodies against growth hormone, prolactin, luteinizing hormone β-subunit, basic cytokeratin (bCK), and S100B. The culture cells on the PLL- and laminin-coated surfaces were round or oval in shape, and bCK-immunopositive FS cells were morphologically indistinguishable from endocrine cells. In the collagen-coated plate, many endocrine cells were round or oval in shape, but FS cells displayed a larger and flattened morphology. S100B-immunoreactions were localized in the nuclei of bCK-immunopositive FS cells. These results suggest that culturing the chicken adenohypophyseal cells in the collagen-coated plate enables the distinction of FS cells from endocrine cells.
将鸡腺垂体细胞培养在涂有不同材料的培养皿中,并检查其形态,以确认鸡卵泡星状(FS)细胞在体外的特征。腺垂体细胞在培养基中用胶原酶/胰蛋白酶混合物分散,并接种到涂有聚-L-赖氨酸(PLL)、胶原或层粘连蛋白的培养皿中。培养7天后,固定细胞并进行免疫细胞化学检测。5-溴-2'-脱氧尿苷掺入试验表明,培养细胞的增殖活性因包被材料而异,在胶原包被的培养皿中增殖活性高于其他两种包被材料。还使用针对生长激素、催乳素、黄体生成素β亚基、碱性细胞角蛋白(bCK)和S100B的混合抗体进行荧光免疫细胞化学检测。在PLL和层粘连蛋白包被表面的培养细胞呈圆形或椭圆形,bCK免疫阳性的FS细胞在形态上与内分泌细胞无法区分。在胶原包被的培养皿中,许多内分泌细胞呈圆形或椭圆形,但FS细胞呈现出更大且扁平的形态。S100B免疫反应定位于bCK免疫阳性FS细胞的细胞核中。这些结果表明,将鸡腺垂体细胞培养在胶原包被的培养皿中能够区分FS细胞和内分泌细胞。