Cyrino J E, Mulvaney D R
Departamento de Produção Animal, ESALQ-USP, C.P. 09, Piracicaba, SP, Brazil.
Rev Bras Biol. 1999 Aug;59(3):517-25. doi: 10.1590/s0034-71081999000300017.
Bioassays were performed to assess the effects of different levels of growth medium supplementation with fetal bovine serum (FBS), fish fry extract (FE), combinations of FBS and FE, and addition of insulin-like growth factor I (IGF-I) and fibroblast growth factor (FGF) on the proliferation of brown bullhead catfish cells (BB line). Treatments (n = 4) were: 2.5, 5, 10, and 15.0% FBS or FE and 5/2.5, 5/5, 10/2.5, and 10/5 of a FBS/FE combination as supplement to the growth medium, or the addition of 0.1, 1, 2.5, 10, 25, and 75 ng/ml of either IGF-I or FGF to the growth media. Initial cell density was 1.1 x 10(6) cells per well on uncoated 24-well plates. Incubation temperature was 29.5 +/- 0.7 degrees C. Six hours after plating, initial culture medium was removed, plates rinsed with Dulbecco's phosphate buffered saline, treatment media added, and cells allowed to proliferate for 24 hours. Another bioassay was performed with rat myoblast omega cells (RMo) using the same levels of growth medium supplemented with FBS, FE and FBS/FE. Base growth medium was Dulbecco's MEM. The initial cell density was 7.2 x 10(6) cells per well, and the bioassay was carried out at 36.0 +/- 0.5 degrees C, on a 95% air, 5% CO2 incubator. Increasing levels of FBS had a positive effect (P < 0.05) on the proliferation of both BB and RMo cells. Increasing levels of FE had a negative effect (P < 0.05) on the proliferation of BB cells and totally inhibited the proliferation of RMo cells at any level of supplementation. Higher levels of FE on the FBS/FE combinations presented a negative effect on the proliferation of both BB and RMo cells (P < 0.05). Insulin-like growth factor I had a positive quadratic effect (P < 0.05) on the proliferation of BB cells. Apparently, mammalian growth factors slightly stimulated mitogenic activity in fish cells, while FE contained factors which inhibited the mitogenic activity of RMo and BB cell lines.
进行了生物测定，以评估不同水平的添加胎牛血清（FBS）、鱼苗提取物（FE）、FBS与FE的组合以及添加胰岛素样生长因子I（IGF-I）和成纤维细胞生长因子（FGF）的生长培养基对褐首鲶细胞（BB系）增殖的影响。处理组（n = 4）为：2.5%、5%、10%和15.0%的FBS或FE，以及5/2.5、5/5、10/2.5和10/5的FBS/FE组合作为生长培养基的补充剂，或者向生长培养基中添加0.1、1、2.5、10、25和75 ng/ml的IGF-I或FGF。初始细胞密度为每孔1.1×10⁶个细胞，接种于未包被的24孔板中。孵育温度为29.5±0.7℃。接种6小时后，去除初始培养基，用杜氏磷酸盐缓冲盐水冲洗平板，加入处理培养基，使细胞增殖24小时。使用相同水平添加FBS、FE和FBS/FE的生长培养基对大鼠成肌细胞ω细胞（RMo）进行了另一项生物测定。基础生长培养基为杜氏MEM培养基。初始细胞密度为每孔7.2×10⁶个细胞，生物测定在36.0±0.5℃的95%空气、5%二氧化碳培养箱中进行。FBS水平的增加对BB和RMo细胞的增殖有积极影响（P < 0.05）。FE水平的增加对BB细胞的增殖有负面影响（P < 0.05），并且在任何补充水平下都完全抑制了RMo细胞的增殖。FBS/FE组合中较高水平的FE对BB和RMo细胞的增殖均有负面影响（P < 0.05）。胰岛素样生长因子I对BB细胞的增殖有正二次效应（P < 0.05）。显然，哺乳动物生长因子对鱼类细胞的有丝分裂活性有轻微刺激作用，而FE含有抑制RMo和BB细胞系有丝分裂活性的因子。
