实时观察Rho激酶对离体培养施万细胞形态和迁移的作用

Role of Rho kinase in morphology and migration of cultured Schwann cells based on time-lapse imaging

目的探讨Rho激酶(ROCK)对离体培养大鼠施万细胞形态和迁移的作用。方法运用原代培养施万细胞技术,并采用细胞免疫荧光化学技术鉴定施万细胞的纯度,同时建立实时观察施万细胞形态和迁移的单细胞迁移平台。基于该平台,在迁移施万细胞前方给予ROCK抑制剂Y-27632浓度梯度,显微实时观察该抑制剂对施万细胞形态和迁移的作用。结果 1.细胞免疫荧光化学显示原代培养的细胞为高纯度的施万细胞;2.单细胞迁移平台非常适合实时研究施万细胞形态和迁移;3.在迁移施万细胞前方给予ROCK抑制剂Y-27632浓度梯度能明显地促进施万细胞突起分支增多,突起变细长,并抑制施万细胞迁移。结论 ROCK能维持施万细胞形态,并调节施万细胞运动、迁移能力。

Objective To investigate the effect of Rho kinase （ROCK） on the morphology and migration of cultured Schwann cells. Methods Primary culture Schwann cells were performed and the purity of Schwann cell was identified by immunofluorescenee. Single-cell migration assay for Schwann cells was established. Based on this assay, a gradient of ROCK inhibitor Y-27632 was produced in front of migrating Sehwann cells. The behavior of morphology and motility in Schwann cells by Y-27632 were recorded based on time-lapse imaging. Results Immunofluorescence showed that cultured cells were of high purity of Schwann cell and single-cell migration assay was suitable for studying the morphology and motility of Schwann cells. Y-27632 dramatically promoted the more braehes, thinner and longer processes, and inhibited Sehwann cell migration. Conclusion ROCK can maintain the morphology and regulate the motility of Schwann cells.