目的探讨犬上皮细胞原代培养的可行性,并从中分选出一类上皮细胞亚群。方法酶消化法获取121腔黏膜上皮细胞,流式分选整合素a6+/CD71-的细胞亚群,观察其生物学特性,对照组细胞未用流式分选。结果成功获取犬上皮细胞,细胞亚群形态一致,可见角蛋白丝和桥粒结构,克隆形成率(11.63%)高于对照组(6.83%),生长周期也更长;第3代亚群细胞整合素仅6阳性表达率(92.3%)高于对照组(86.4%),且整合素B1荧光强度也明显增强。结论犬上皮细胞原代培养成功,并分选出一种新型、优质的种子细胞。
Objective To explore the method of primary culture of canine epithelial cells, and to sort a subset of cell groups. Methods Oral mucosa was dissociated into single cell suspensions with enzyme. Cell subsets (integrin a6 +/CDT1-) were isolated by using flow eytometry, and their biological characteristics were observed. The control group was not sorted by flow cytometry. Results Epithelial cells were successfully isolated and cultured. Cell subsets had similar morphological characteristics, and keratin filaments and desmosome structure could be seen. Compared with the control group (6. 83% ) , clone for- mation rate of cell subpopulations was 11.63 % , and cell supopulations also showed a longer growth period. Integrin c~6 expression rate (92. 3% ) in the third generation of cell subsets was higher than that in the con- trol group (86. 4% ), and integrin 131 fluorescence intensity was significantly higher too. Conclusion Ca- nine epithelial cells could be successfully obtained by primary culture, and we sorted out a kind of new and good seeding cells.