大鼠胰腺导管上皮细胞体外诱导分化

Inducing rat pancreatic ductal epithelial cells to differentiate into islet cells in vitro

目的探讨上皮细胞生长因子(EGF)、尼克酰胺(NIC)、肝细胞生长因子(HGF)、葡萄糖(Glu)、角脘蛋白生长因子(KGF)和β细胞素在大鼠胰腺导管上皮细胞体外诱导分化的作用。方法采用健康SD大鼠胰腺导管上皮细胞,分于70个培养孔中,加入含有多种营养因子(不含血清)的PRMI1640培养液中培养。每个孔中6种诱导剂均进行随机组合。在胰岛样细胞团(ICCs)形成前后的不同天数进行细胞计数、电镜观察、胰岛素测定以及免疫细胞化学和荧光分析。结果胰腺导管上皮细胞培养7d后,单个贴壁上皮细胞分裂增殖,呈鹅卵石样细胞片,单层覆盖;加入诱导和促生长因子后,单层覆盖上皮细胞中逐渐产生一定数量类似胰岛样球状细胞团,直径为80～130μm。结论6种促生长和诱导因子中,Glu、NIC、KGF、EGF作用较强;在所设定的浓度范围内,其作用均随着浓度的增加而增强。

Objective To study the role of epidermal growth factor（EGF）, hepatocyte growth factor（HGF）, β-cellulin, nicotinamide, keratinocyte growth factor （KGF） and glucose （Glu） in differentiation of rat pancreatic β-cells in-vitro. Methods Duct epithelial cells were cultivated in serum-free RPMI 1640 medium with the addition of nutrition factors and distributed randomly to 70 holes of petri dish. In the period of islet-like cell clusters（ICCs） formation,we measured the quantity and function of insulin-producing cells by light and electron microscopy, cytometry, basal secreted insulin measurement, and immunocytochemistry. Results After 7 days of cultivation, single epithelial cell adhered to the dish surface extended to monolayers by a characteristic ＂cobblestone＂ appearance. Many ICCs emerged among the monolayers of epithelial cells after adding growth factors and inducers. Each diameter was about 80-130μm. Conclusion Amoung 6 factors and inducers, the effects of NIC,KGF, EGF and Glu were stronger and concentration-dependent.