Seaver S S, van der Bosch J, Sato G
Exp Cell Res. 1984 Nov;155(1):241-51. doi: 10.1016/0014-4827(84)90785-7.
A simple three-enzyme treatment of collagenase, dispase and hyaluronidase on finely minced chick oviduct yields clumps of 50-150 cells. These cells attach to collagen-treated dishes and survive in culture for at least 2 weeks without subculturing. Oviduct cell cultures can also be induced to grow. Estradiol or epidermal growth factor (EGF) induce a 40% increase in cells in 4 days when cultures are grown in serum levels that do not support growth. Serum from estrogen-stimulated chicks promotes rapid cellular proliferation (doubling times of 1-2 days). Sera from estrogen withdrawn chicks, laying hen or horse do not support as rapid proliferation. The oviduct growth-promoting factors in serum from estrogen-stimulated chicks are not steroids or fibroblast growth factors (FGF). Removal of steroids from these sera by charcoal treatment or delipidization does not decrease the rate of growth. The addition of 1-100 nM estradiol does not increase a serum's ability to promote growth. Purified FGF or platelet-derived growth factor (PDGF) do not induce oviduct proliferation. These results were reproduced in oviduct cell cultures started from estrogen-stimulated and withdrawn chicks as well as laying hens. Thus the factors in serum from estrogen-stimulated chicks that promote rapid oviduct growth are induced by estrogen treatments in vivo, but do not seem to be only steroids.
用胶原酶、分散酶和透明质酸酶对精细切碎的鸡输卵管进行简单的三酶处理,可得到50 - 150个细胞的团块。这些细胞附着在经胶原处理的培养皿上,在不进行传代培养的情况下可在培养中存活至少2周。输卵管细胞培养物也可被诱导生长。当培养物在不支持生长的血清水平中生长时,雌二醇或表皮生长因子(EGF)在4天内可使细胞数量增加40%。来自雌激素刺激的小鸡的血清可促进细胞快速增殖(倍增时间为1 - 2天)。来自雌激素撤除的小鸡、产蛋母鸡或马的血清则不支持如此快速的增殖。来自雌激素刺激的小鸡血清中的输卵管生长促进因子不是类固醇或成纤维细胞生长因子(FGF)。通过活性炭处理或脱脂从这些血清中去除类固醇并不会降低生长速率。添加1 - 100 nM的雌二醇不会增加血清促进生长的能力。纯化的FGF或血小板衍生生长因子(PDGF)不会诱导输卵管增殖。这些结果在从雌激素刺激和撤除的小鸡以及产蛋母鸡开始的输卵管细胞培养中得到了重现。因此,来自雌激素刺激的小鸡血清中促进输卵管快速生长的因子是由体内雌激素处理诱导产生的,但似乎不仅仅是类固醇。