孔源性视网膜脱离患者视网膜表面膜组织的体外培养

Isolation and culture of epiretinal membranes from patients withrhegmatogenous retinal detachment in vitro

目的 建立体外培养增生性玻璃体视网膜病变(PVR)患者病变膜组织的方法,为提供PVR研究奠定基础。方法 孔源性视网膜脱离伴有PVR分级D级患者进行玻璃体切割术同时获取其视网膜表面膜病变(ERM)标本,患者病程7－120天;ERM膜片应用切块贴皿法进行接种,培养皿短时间斜置使ERM组织贴壁,每3－4天换培养液1次,观察记录膜组织中细胞的迁出、分裂和增生状况,部分病变标本进行免疫细胞化学鉴定。结果7例患者的ERM组织提供体外原代培养,培养期间膜片色素变淡,其中3例患者的膜组织有细胞从膜片边缘向外迁移,生长缓慢,迁出的细胞呈现两种形态:铺路石样和梭形。cytokeratin染色呈阳性。结论 患者ERM体外培养方法可行,细胞生长情况与多因素有关。ERM中的细胞主要是RPE细胞,其增殖能力有可能评估手术预后。

Objective To develop the techniques of culture in vitro for epiretinal membrane(ERM) tissues from patients with proliferative vitreoretinopathy(PVR). Methods All specimens were surgically removed by vitrectomy in the patients with D grade PVR in rhegmatogenous retinal detachment. The duration of retinal detachment was 7 to 120 days. The tissue pieces were cultured using microdissection techniques on 3. 5 mm dishes. The cultures were observed daily with inverted phase microscopy. Some were studied further by immunocytochemistry. Results 7 ERMs were obtained for primary culture. The pigment of tissues began to fade after 24 hours. Cells migrated from the explants in three specimens, but the cells were hard to attain confluence. The cells,which grew from the explants, showed two variable appearances including cobblestone-like and spindle-shaped cells. The staining for cytokeratin in these cells was positive. Conclusion Our study suggests that culture of ERMs in vitro is feasible. The condition of the cell growth possibly allows coupling with prognosis, and the propensity of cell migration and growth, however, depends on duration and operated techniques for ERMs. RPE cells are the predominant cell types in ERM from PVR.