论文部分内容阅读
目的:研究SOX9的表达在诱导肝细胞向胰腺干祖样细胞转化过程中的意义。方法:分离培养小鼠原代肝细胞,包装、浓缩表达OCT4的慢病毒并感染肝细胞,使用RT-PCR监测慢病毒感染后肝细胞中SOX9的表达,并根据SOX9表达情况尝试向胰腺干祖样细胞诱导。使用PCR和免疫荧光鉴定生成的胰腺干祖样细胞。结果:肝细胞感染OCT4-慢病毒后,肝细胞内颗粒逐渐释出,细胞折光性增强。SOX9表达从第6 d持续至第8 d。从SOX9阴性的第4 d开始向胰腺诱导,无集落样的胰腺干祖样细胞生成,RT-PCR检测其标志物CK19为阴性;从SOX9阳性的第6 d开始向胰腺诱导,可见集落样生长的胰腺干祖样细胞,RT-PCR检测CK19为阳性,免疫荧光染色显示CK19表达在胞浆中。结论:肝细胞向胰腺干祖样细胞转化过程中SOX9的表达提示了“兼性肝细胞”的出现,此细胞具有向胰腺谱系诱导分化的潜能,这为进一步生成有功能的胰岛内分泌细胞和实现糖尿病的细胞替代治疗提供了理论和实验依据。
Objective: To investigate the significance of SOX9 expression in inducing hepatocytes to transform into pancreatic stem-progenitor cells. Methods: Primary hepatocytes of mice were isolated and cultured. The lentivirus expressing OCT4 was concentrated and infected into hepatocytes. The expression of SOX9 in hepatocytes was detected by RT-PCR. According to the expression of SOX9, Like cell induction. The generated pancreatic stem-progenitor cells were identified by PCR and immunofluorescence. Results: After the hepatocytes were infected with OCT4-lentivirus, the intragranular granules gradually released and the refraction of cells increased. SOX9 expression continued from the 6th day to the 8th day. From the fourth day after SOX9-negative induction, the pancreatic stem-like progenitor cells were induced to grow without colony formation, and CK19 was negative by RT-PCR. From the sixth day after SOX9-positive induction, Of pancreatic stem-progenitor cells, positive for CK19 detected by RT-PCR, immunofluorescent staining showed CK19 expression in the cytoplasm. CONCLUSIONS: The expression of SOX9 during the transformation of hepatocytes into pancreatic stem-progenitor cells suggests the emergence of “facultative hepatocytes”, which have the potential to induce differentiation into the pancreatic lineage, which further contributes to the generation of functional islet endocrine cells And to provide a theoretical and experimental basis for the replacement therapy of diabetes cells.