Collodi P, Barnes D W
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331-6503.
Proc Natl Acad Sci U S A. 1990 May;87(9):3498-502. doi: 10.1073/pnas.87.9.3498.
An extract of 21-day rainbow trout embryos stimulated growth of several piscine cell lines in the absence of added serum. Established lines from trout (RTG-2 and STE-137), salmon (CHSE-214), carp (EPC), and goldfish (CAR) and early-passage cells initiated from trout embryos grew in serum-free medium containing the embryo extract. In addition the extract was sufficient for maintaining long-term cultures of CHSE-214 cells for several months through a minimum of 20 passages (approximately 50 population doublings) in the absence of serum. Optimal response was achieved with 100 micrograms of extract protein per ml, but a significant growth-promoting effect was observed with as little as 2.5 micrograms/ml. The activity was nondialyzable, protease-sensitive, and stable in 200 mM acetic acid. The level of mitogenic response induced by the extract could not be duplicated with purified mammalian growth factors added individually or in combination, and the extract did not stimulate DNA synthesis in quiescent mouse fibroblasts. These results suggest that trout embryo extract may contain a novel growth-promoting activity for fish cells.
21日龄虹鳟胚胎的提取物在不添加血清的情况下刺激了几种鱼类细胞系的生长。来自虹鳟(RTG-2和STE-137)、鲑鱼(CHSE-214)、鲤鱼(EPC)和金鱼(CAR)的已建立细胞系,以及从虹鳟胚胎起始的早期传代细胞,在含有胚胎提取物的无血清培养基中生长。此外,该提取物足以在无血清的情况下通过至少20次传代(约50次群体倍增)维持CHSE-214细胞的长期培养数月。每毫升100微克提取物蛋白可实现最佳反应,但低至2.5微克/毫升时也观察到显著的促生长作用。该活性不可透析,对蛋白酶敏感,且在200 mM乙酸中稳定。提取物诱导的促有丝分裂反应水平不能通过单独或组合添加纯化的哺乳动物生长因子来复制,并且该提取物不会刺激静止的小鼠成纤维细胞中的DNA合成。这些结果表明,虹鳟胚胎提取物可能含有一种对鱼类细胞具有新型促生长活性的物质。