BXSB狼疮小鼠胸腺上皮细胞培养方法的建立

Establishment of thymic epithelial cell culture system of BXSB systemic lupus erythematosus mice

目的 :比较不同培养条件下原代狼疮小鼠胸腺上皮细胞生长情况。方法 :分别采用剪切法、剪切加胶原酶消化法、剪切加胰蛋白酶消化法建立培养体系获取自发性系统性红斑狼疮小鼠 (BXSB小鼠 )胸腺上皮细胞 ;用光镜、免疫细胞化学染色法进行细胞鉴定。结果 :含血清的培养基与含生长因子的培养基均能培养出较纯的胸腺上皮细胞 ;剪切法成纤维细胞污染较多 ,剪切加胰蛋白酶消化法细胞生长状态较差 ,经剪切加胶原酶消化的BXSB胸腺植块 ,生长状态好 ,成纤维细胞污染少 ;当原代培养的细胞长满瓶壁的 90 %左右时可传代 ,角蛋白染色阳性细胞纯度达 95 %以上。结论 :剪切加胶原酶消化法仅用含血清的培养基即可培养出符合实验要求的 BXSB小鼠胸腺上皮细胞 ,是低成本、简便易行的系统性红斑狼疮模型鼠胸腺上皮细胞培养体系。

Objective To study and establish the culture system of thymic epithelial cell of BXSB systemic lupus erythematosus mice. Methods The thymic epithelial cells of systemic lupus erythematosus model (BXSB mice) were cultured with merely shearing, shearing and collagenase digestion, shearing and trypsinization in different nutrient media . The cells were identified with microscopy and immunohistochemical methods. Results The pure thymic epithelial cells were obtained not only in the medium with serum but also in the medium with growth factor. The thymic epithelial cells grew very well with shearing and collagenase digestion, and there was seldom contaminated by fibroblast. Passage was operated when the primary culture cells spreaded more than 90% pavement of culture flask. The purity of cells dyed with keratin was more than 95% in the medium with serum by shearing and collagenase digestion. Conclusion The method of shearing and collagenase digestion process is easy and cheap to obtain thymic epithelial cell of BXSB mice. A stable thymic epithelial cell culture system of BXSB systemic lupus erythematosus mice is established.