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目的:探讨胎牛血清对小鼠角膜上皮细胞生长和分化的影响。方法:分别在无血清低钙培养基(KSFM)和含100mL/L胎牛血清的KSFM中培养小鼠角膜上皮细胞。比较两种培养基中角膜上皮细胞的群体倍增(PD)。通过RT-PCR和Western blotting方法检测p63、角蛋白19以及involucrin的表达。结果:在KSFM中,小鼠角膜上皮细胞可稳定传代超过25代,但在含胎牛血清的培养基中,细胞仅能存活3代。在KSFM中,培养细胞呈典型铺路石样外观,RT-PCR和Western blotting结果显示细胞表达p63、角蛋白19以及in-volucrin。当培养基中加入胎牛血清后,细胞形态明显增大,呈扁平状; involucrin表达显著增强。结论:胎牛血清可抑制小鼠角膜上皮细胞增生、诱导其分化。
Objective: To investigate the effects of fetal calf serum on the growth and differentiation of mouse corneal epithelial cells. Methods: Mouse corneal epithelial cells were cultured in serum free low calcium medium (KSFM) and KSFM containing 100 mL / L fetal bovine serum respectively. Corneal epithelial cell population doublings (PD) were compared in both media. The expression of p63, keratin 19 and involucrin were detected by RT-PCR and Western blotting. Results: In KSFM, mouse corneal epithelial cells can be passaged more than 25 passages steadily, but in medium containing fetal bovine serum, cells can only survive for 3 passages. In KSFM, the cultured cells showed a typical paving stone-like appearance. RT-PCR and Western blotting showed that the cells expressed p63, keratin 19, and in-volucrin. When fetal bovine serum was added into the culture medium, the cell morphology was significantly increased, flat; involucrin expression was significantly enhanced. Conclusion: Fetal calf serum can inhibit mouse corneal epithelial cell proliferation and induce its differentiation.