摘要
目的探讨大鼠胰腺提取物(RPE)是否可以使小鼠骨髓间充质干细胞(BMMSCs)分化为胰岛素产生细胞(IPCs),以及小鼠BMMSCs的分化条件,为糖尿病的干细胞治疗提供新的方法。方法传代纯化培养昆明小鼠的骨髓间充质干细胞,应用SD大鼠的RPE诱导分化小鼠BMMSCs为IPCs,首先取18瓶小鼠BMMSCs随机分为6组,每组3瓶,分别加入RPE 10mg/L、20mg/L、30mg/L、40mg/L、50mg/L、100mg/L,筛选出最佳的分化剂量,然后用最佳的分化剂量(50mg/L)分化BMMSCs,免疫酶细胞化学鉴定细胞内胰岛素的表达;双硫腙染色检测胰岛素产生细胞的分化和纯度。放射免疫分析检测C肽片段表达。结果50mg/L RPE可以使间充质干细胞分化良好,形态较规则,1周后细胞分化成熟,双硫腙染色呈现猩红色;免疫酶细胞化学和免疫荧光细胞化学显示:胰岛素表达阳性,C肽放射免疫学显示有C肽释放。结论RPE可以使小鼠的骨髓间充质干细胞分化为胰岛素产生细胞。
Objective To research the method of mice bone marrow mesenchymal stem cells （BMMSCs） trans- differenting into insulin producing cells （IPCs） by rat pancreatic extract （RPE）. Methods Mesenchymal stem cells were isolated from Kunming mouse, and RPE was from SD rats＇ pancreases. Passaged BMMSCs were induced to be insulin-producing cells（IPCs） with RPE. First, derided mice＇s BMMSCs to 6 groups,3 culture flasks per group at random, and added RPE 10mg/L, 20mg/L, 30mg/L, 40mg/L, 50mg/L, 100mg/L to each group. After 1 week, decided which concentration of RPE was better. And induce mouse BMMSCs to IPCs by it（50mg/L）. The expression of insulin in IPCs was detected by immunoenzyme cytochemistry and immunofluorescence cytochemistry, the trans-differentiation purity of IPCs was detected by dithizone （DTZ）-stain,C-peptide secreting was detected by radioimmunoassay（RIA）. Results Mesenchymal stem cells of mice could be induced into IPCs by 50mg/L PRE. Typical islet-like clustered cells were observed after lweeks, and could be stained scarlet by DTZ. The insulin expression of BMMSCs was positive by immunoenzyme cytochemistry and immunofluorescence cytochemistry. RIA showed the expression of C-peptide in the IPCs could be detected. Conclusion RPE could induce BMMSCs of mice to differenciate into IPCs.
出处
《解剖学报》
CAS
CSCD
北大核心
2009年第5期789-793,共5页
Acta Anatomica Sinica
