Thordarson G, Folger P, Talamantes F
Thimann Laboratories, University of California, Santa Cruz 95064.
Placenta. 1987 Nov-Dec;8(6):573-85. doi: 10.1016/0143-4004(87)90028-2.
Fetal placental tissue from 11 days pregnant mice was dissociated in collagenase and DNase solution and then separated on a 40 per cent Percoll gradient. Trophoblast cells banded at a density of 1.05 g/ml. When cultured on rat tail collagen, these cells formed colonies of mono- and binucleate cells varying in size from 40 to 70 microns. At the time of plating, about 13 per cent of the trophoblast cells secreted mouse placental lactogen II (mPL-II) as determined by reverse haemolytic plaque assay. The ratio of mPL-II-producing cells increased significantly in culture and reached 63 per cent after 48 h. The secretion of mPL-II increased continuously during six days of culture, whereas the total protein release was highest after the first day, declined the second day and then remained relatively constant for the last four days of culture. The DNA content of the cells did not change significantly during the six-day period. When the trophoblast cells were incubated with insulin (1 ng/ml to 5 micrograms/ml), a modest but significant reduction in mPL-II secretion was observed. No change in the mPL-II secretion was seen when epidermal growth factor was administered to the culture in concentrations from 1 ng/ml to 10 micrograms/ml. It is concluded that this in vitro culture system is suitable for studying both mPL-II secretion and the differentiation of mPL-II-producing cells.
将怀孕11天的小鼠的胎盘组织在胶原酶和DNA酶溶液中解离，然后在40%的 Percoll 梯度上进行分离。滋养层细胞在密度为1.05 g/ml处形成条带。当在大鼠尾胶原上培养时，这些细胞形成了大小从40到70微米不等的单核和双核细胞集落。接种时，通过反向溶血空斑试验测定，约13%的滋养层细胞分泌小鼠胎盘催乳素II（mPL-II）。培养过程中，产生mPL-II的细胞比例显著增加，48小时后达到63%。培养六天期间，mPL-II的分泌持续增加，而总蛋白释放量在第一天最高，第二天下降，然后在培养的最后四天保持相对稳定。细胞的DNA含量在六天期间没有显著变化。当滋养层细胞与胰岛素（1 ng/ml至5 μg/ml）一起孵育时，观察到mPL-II分泌有适度但显著的减少。当以1 ng/ml至10 μg/ml的浓度向培养物中加入表皮生长因子时，mPL-II的分泌未见变化。结论是，这种体外培养系统适用于研究mPL-II的分泌以及产生mPL-II的细胞的分化。
