Soonpaa M H, Oberpriller J O, Oberpriller J C
Department of Anatomy and Cell Biology, University of North Dakota School of Medicine, Grand Forks 58202.
J Mol Cell Cardiol. 1992 Sep;24(9):1039-46. doi: 10.1016/0022-2828(92)91870-b.
The adult newt cardiac ventricular myocyte has been successfully placed in cell culture and has been shown to undergo in vitro DNA synthesis. Although several growth factors have been reported to increase DNA synthesis in cardiac myocytes in vitro, PDGF has not been reported to do so, but has been shown to be active in other systems. Ventricles were removed from the adult red-spotted newt and were enzymatically and mechanically dissociated in a solution containing trypsin and collagenase. Cells were preplated on to plastic to remove non-myocytes. Myocytes were then plated onto laminin. Groups of myocytes were fed control medium and medium containing porcine PDGF. Myocytes were given 1 microCi/ml of tritiated thymidine 6 or 24 h before fixation. Control myocytes showed a peak DNA synthesis at 12-14 days in culture. One ng/ml of PDGF increased DNA synthesis significantly to 22% above control. Myocytes responded to PDGF with significantly increased DNA synthesis in about 12 h. PDGF did not induce earlier DNA synthesis, but increased synthesis at all days of culture tested. These results indicate that PDGF acts upon cardiac myocytes, increasing their DNA synthesis.
成年蝾螈的心室肌细胞已成功培养于细胞培养物中，并已证明其可在体外进行DNA合成。虽然有报道称几种生长因子可在体外增加心肌细胞中的DNA合成，但血小板衍生生长因子（PDGF）尚未见有此报道，不过它已被证明在其他系统中具有活性。从成年红斑蝾螈中取出心室，在含有胰蛋白酶和胶原酶的溶液中进行酶解和机械解离。细胞先接种于塑料培养皿上以去除非心肌细胞。然后将心肌细胞接种于层粘连蛋白上。将心肌细胞分组，分别给予对照培养基和含有猪PDGF的培养基。在固定前6或24小时，给心肌细胞加入1微居里/毫升的氚标记胸腺嘧啶核苷。对照心肌细胞在培养12 - 14天时显示出DNA合成高峰。1纳克/毫升的PDGF可使DNA合成显著增加，比对照高出22%。心肌细胞在约12小时内对PDGF做出反应，DNA合成显著增加。PDGF并未诱导更早的DNA合成，但在所有测试的培养天数均增加了合成。这些结果表明，PDGF作用于心肌细胞，增加其DNA合成。
