增殖性玻璃体视网膜病变视网膜前膜体外培养及免疫组化鉴定

In Vitro Culture and Immunohistochemistry Identification of Epiretinal Membranes with PVR

目的探讨视网膜前膜组织培养中存在的细胞迁出率低、传代困难等问题；观察培养细胞的成分及体外培养细胞迁出率和增殖力与PVR发病时间的相关性。方法使用组织块培养法对手术中剥离的PVR患者视网膜前膜组织进行体外培养；双目倒置显微镜下观察细胞生长状况；DAB显色法对细胞进行免疫组化鉴定。结果PVR在3个月内的细胞迁出率与3个月以上的细胞迁出率和细胞传代率都有统计学意义；共培养31例前膜组织其中23例有细胞迁出，14例成功进行了细胞传代，8例传至第四代；免疫组化鉴定见：s-100、CK为阳性，Vimt、GFAP仅少部分细胞呈阳性，CD34呈阴性反应。结论PVR3个月内的患者的视网膜前膜组织是进行视网膜前膜组织体外培养的最佳选材；培养的细胞视网膜色素上皮细胞为主，含有少部分成纤维细胞及神经胶质细胞。

Objective To develop the technique of culture in vitro for ERMs from PVR and identify the cell types.Compare the cell migratory capabilities in different stages of PVR.Methods 31 ERMs specimens were surgically removed by vitrectomy in the patients with D1-D3 grade PVR.The tissue pieces were cultured with DMEM-F12 nutritive in six pore culture plate.The cells were identified with immunohistochemistry staining method.Results The cell migratory capabilities have the significant difference between in 3month and above 3 month for ERMs of PVR（x^2= 3.876 α〈 0.05 ;x^2= 6.626 α〈 0.05 ）. Total culture results show the cell will emigrate from the tissue in 10 days after seeding,there are 23 ERMs specimens with cell emigration, 14specimens with well growth and passage,8 specimens getting the fourth passage with well growth condition.The cells would grow well in metaacid microenvironment.The immunohistochemistry staining show most of cells were positive for S-100 and CK,a few cells were positive for GFAP and Vimt,all cells were negative for CD34.Conclusion The ERMs specimens were the best material for culture in vitro in 3month of PVR.The main type of cells is RPE cell and part of fibroblasts cell,RG cell.