胰酶消化法培养大鼠胸主动脉平滑肌细胞

Culture of Vascular Smooth Muscle Cells From the Rat Thoracic Aorta By Trypsin Digestion Method

建立胰酶消化法培养大鼠胸主动脉平滑肌细胞 ,并和其他大鼠胸主动脉平滑肌细胞的培养方法相比较。在无菌条件下分离雄性Wistar大鼠胸主动脉血管中膜 ,剪碎中膜 ,在 37℃用 10mL 0 .2 5 %胰酶消化大约 3.5h。中止消化后 ,在 10 0g条件下离心 5min ,收集细胞种入 2 5cm2 细胞培养瓶。结果发现 ,胸主动脉平滑肌细胞至少可以传 2 0代以上 ,并且细胞形态、生长特点、平滑肌a 肌动蛋白表达不发生明显的改变。结果提示 ,与目前的平滑肌细胞培养方法相比 ,胰酶消化法培养平滑肌细胞的方法重复性好 ,原代培养的平滑肌细胞具有数量多、生长迅速的特点。

Aim To develop a method of culturing vascular smooth muscle cells (VSMC) from isolated rat thoracic aorta and compare their growth with that of aortic VSMC by other culture method. Methods The vascular media of rat thoracic aorta was isolated from male Wistar rats in sterile condition. Mince the media, and suspend the media in 10 mL 0.25% trypsin for about 3.5 h at 37℃. After terminating the digestion, centrifuge the solution at 100 g for 5 min, and seed the cell into a 25 cm2 vented cap flask. Results Thoracic aorta smooth muscle cell (SMC) were successfully passaged more than 20 times, without noticeable changes in morphology, growth characteristics, and smooth muscle α-actin expression. Conclusions Compared with the currently employed methods, our technique has the advantages of good reproducibility, high yield of pure SMC and quick growth of primary culture.