Lapp C A, Stachura M E, Tyler J M, Lee Y S
Department of Medicine, Medical College of Georgia, Augusta 30910.
In Vitro Cell Dev Biol. 1987 Oct;23(10):686-90. doi: 10.1007/BF02620981.
GH3 cell secretory activity was studied in long-term perifusion to define previously reported spontaneous increases in growth hormone (GH) and prolactin production (PRL). Mechanically harvested cells (1 X 10(7)/column) were perifused at 4 ml/h for 72 h. A basal period of variable duration (8 to 12 h), during which hormone secretion was stable, was followed by steadily increasing secretion rates. Changes in cell number were not sufficient to account for increased hormone secretion rates: a) there was no significant change in cell count after 72 h (0.97 +/- 0.03 X 10(7); n = 18); b) mean cell column DNA content increased 25.5% above the base value, whereas GH secretion rose 385% and PRL rose 178% (n = 5). Observed differences in the duration of the basal secretion period, the basal secretory rate, and the magnitude of secretory rate increase were associated with several variables: a) variability within a subline was a function of passage number: GH secretion decreased and PRL secretion increased with subculture number; b) cells with identical lot and freeze numbers, but received at different times, behaved differently; c) the presence of an antifungal agent (nystatin) altered hormone secretion reproducibly.
a) rates of GH and PRL secretion rise spontaneously in perifusion without a proportional increase in GH3 cell number; b) fluctuations in the rate of GH3 cell secretion of GH and PRL are not entirely random but are determined by several definable variables.
通过长期灌流研究GH3细胞的分泌活性,以确定先前报道的生长激素(GH)和催乳素分泌(PRL)的自发增加。机械收获的细胞(1×10⁷/柱)以4 ml/h的速度灌流72小时。在持续时间可变的基础期(8至12小时),激素分泌稳定,随后分泌率稳步增加。细胞数量的变化不足以解释激素分泌率的增加:a)72小时后细胞计数无显著变化(0.97±0.03×10⁷;n = 18);b)平均细胞柱DNA含量比基础值增加了25.5%,而GH分泌增加了385%,PRL分泌增加了178%(n = 5)。观察到的基础分泌期持续时间、基础分泌率和分泌率增加幅度的差异与几个变量有关:a)亚系内的变异性是传代次数的函数:随着传代次数的增加,GH分泌减少,PRL分泌增加;b)批号和冻存编号相同但接收时间不同的细胞表现不同;c)抗真菌剂(制霉菌素)的存在可重复性地改变激素分泌。
a)在灌流过程中,GH和PRL的分泌率自发升高,而GH3细胞数量没有成比例增加;b)GH3细胞分泌GH和PRL的速率波动并非完全随机,而是由几个可定义的变量决定。