大鼠结膜杯状细胞的体外培养及鉴定

发布时间：2018-04-23 05:08

本文选题：结膜杯状细胞 + 细胞培养　；参考：《中国老年学杂志》2017年07期

【摘要】：目的体外分离、培养大鼠结膜杯状细胞,观察其形态、结构等生物学特征。方法取大鼠穹隆部结膜,加入含10%胎牛血清的RPMI 1640培养液中行组织块培养,5~7 d后酶消化法传代。观察原代及传代细胞的形态及生长情况,并应用特殊免疫组化染色方法分别对原代、传代培养的杯状细胞进行鉴定,用Image J软件计算细胞纯度。结果倒置相差显微镜下可见杯状细胞呈鹅卵石样,5~7 d在组织块周围形成环状结节。免疫组化结果显示,培养的原代细胞角蛋白(CK)4染色阴性,CK7染色阳性,且CK7阳性细胞数占(97.3±2.0)%。传代杯状细胞中,绝大多数凝集素UEA-I染色为阳性,阳性细胞占(91.2±2.7)%。结论大鼠穹窿部结膜可以成功培养出杯状细胞,传代细胞仍保持原代细胞特性,且纯度较高。
[Abstract]:Objective to isolate and culture rat conjunctival goblet cells in vitro and observe their morphological and structural characteristics. Methods the conjunctiva of rat fornix was cultured in RPMI 1640 medium containing 10% fetal bovine serum for 5 days and then subcultured by enzyme digestion. The morphology and growth of primary and passage cells were observed, and the primary and cultured goblet cells were identified by special immunohistochemical staining. The purity of the cells was calculated by Image J software. Results under inverted phase contrast microscope, goblet cells were found to form circular nodules around the tissue mass on the 7th day as cobblestone. The results of immunohistochemistry showed that the primary cultured cells were positive for CK7, and the number of CK7 positive cells accounted for 97.3 卤2.0%. Most of the goblet cells were positive for lectin UEA-I, and the positive cells accounted for 91.2 卤2.7. Conclusion the rat fornix conjunctiva can culture goblet cells successfully.
【作者单位】：山西医科大学第二医院眼科;山西医科大学第二医院耳鼻喉科;
【基金】：山西省回国留学人员科研资助项目(2016-062)
【分类号】：R77

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