CAJ | 학술논문

目的: 探讨人骨髓源干细胞向具有功能的胰岛素分泌细胞分化的可能性.方法: 从人骨髓分离间充质干细胞.采用表皮生长因子、β-巯基乙醇和高糖培养基诱导其向胰岛素分泌细胞分化.经诱导后,用RT-PCR检测胰岛β细胞相关基因的表达,并采用免疫细胞化学染色检测胞浆胰岛素的表达.此外,诱导后细胞分泌的胰岛素定量及胰岛素释放实验将采用化学发光法进行检测.将经诱导后的细胞移植到糖尿病小鼠的右侧肾被膜下.在移植后16 d持续检测小鼠的血糖水平,最后对右侧肾脏进行免疫组化检测.结果: 经诱导后,细胞能表达胰岛β细胞相关基因;免疫细胞化学染色也能检测到胞浆有胰岛素的表达;而且这些细胞能对糖刺激有所反应.细胞被移植到糖尿病小鼠的肾被膜下,能起降血糖作用.其肾脏的免疫组化显示:肾被膜下有胰岛素阳性细胞.结论: 人骨髓源干细胞具有向胰岛素分泌细胞分化的潜能,这将为糖尿病细胞治疗提供丰富的细胞来源.
목적: 탐토인골수원간세포향구유공능적이도소분비세포분화적가능성.방법: 종인골수분리간충질간세포.채용표피생장인자、β-구기을순화고당배양기유도기향이도소분비세포분화.경유도후,용RT-PCR검측이도β세포상관기인적표체,병채용면역세포화학염색검측포장이도소적표체.차외,유도후세포분비적이도소정량급이도소석방실험장채용화학발광법진행검측.장경유도후적세포이식도당뇨병소서적우측신피막하.재이식후16 d지속검측소서적혈당수평,최후대우측신장진행면역조화검측.결과: 경유도후,세포능표체이도β세포상관기인;면역세포화학염색야능검측도포장유이도소적표체;이차저사세포능대당자격유소반응.세포피이식도당뇨병소서적신피막하,능기강혈당작용.기신장적면역조화현시:신피막하유이도소양성세포.결론: 인골수원간세포구유향이도소분비세포분화적잠능,저장위당뇨병세포치료제공봉부적세포래원.
AIM: To explore the possibility of differentiating functional insulin-producing cells from human BM-derived stem cells. METHODS: Mesenchymal stem cells were isolated from human bone marrow. Then these cells were induced with epidermal growth factor, β-mercaptoethanol and high concentration of glucose. The gene expression related to islet β cells was detected by RT-PCR. Insulin in the treated cells was examined by immunocytochemistry. In addition, the levels of insulin secretion and glucose-stimulated insulin release were examined by microparticle enzyme immunoassay. Finally, the induced cells were implanted into the right renal subcapsular space of diabetic mice. Blood glucose levels were monitored 16 d after implantation. The right kidneys of the treated mice were harvested for immunohistochemistry. RESULTS: The key genes related to pancreatic β cells had been confirmed to express by PCR and insulin was detected by immunocytochemistry in differentiated human BM-derived stem cells induced by high glucose, which responded to glucose challenge. Furthermore, implantation of the cells in renal subcapsular space was able to lower the glucose levels in hyperglycemic mice. After 16 days, the implanted cells were determined still to be insulin positive cells by immunohistochemistry. CONCLUSION: These results indicate human BM-derived stem cells are capable of differentiating into functional insulin-producing cells and may represent a pool of cells for the treatment of diabetes.