豚鼠耳蜗螺旋动脉平滑肌细胞的原代培养及鉴定.

肖靖杰; 张治平; 王 洋; 司 超; 张 亮; 张 伟; 李 丽; 李新芝; 马克涛

Objective To obtain vascular smooth muscle cells (VSMCs) from spiral modiolar artery of guinea pigs with a combination of two common cultural methods so as to provide materials for related experimental studies. Methods Spiral modiolar artery was separated from the cochlea of guinea pigs first and then digested with 1.25g/L of trypsin to be allowed to adhere. It was then purified by comprehensive application of natural purification and differential adherence ability. Finally, the cells were confirmed by morphological observation, immunofluorescence and immunocytochemistry analysis, as well as Western blot. Results 85% of the explants survived. The smooth muscle cells at passage 4 had a purity of over 95%. The cells were observed to present a typical “hill-and-valley” growth pattern under the microscope. Moreover, immunofluorescence, immunohistochemical staining and Western blot showed the specific and intense positive expression of α-SM-actin, a cell type-specific marker. Conclusion By this protocol, we can obtain a variety of highly purified vascular smooth muscle cells from the spiral modiolar artery of guinea pigs, providing an ideal experiment material for studies on the pathogenesis of diseases related to the circulation disturbance of the inner ear in vitro, as well as evaluation of drug's effect on vascular smooth muscle.

Journal of Xi'an Jiaotong University (Medical Sciences), 2018, Vol 39, Issue 6, p917

1671-8259

Academic Journal

10.7652/jdyxb201806029