AIM: To explore the possibility of marrow mesenchymal stem cells (MSC) in vitro differentiating into functional isletlike cells and to test the diabetes therapeutic potency of Islet-like cells. METHODS: Rat MSCs were ...AIM: To explore the possibility of marrow mesenchymal stem cells (MSC) in vitro differentiating into functional isletlike cells and to test the diabetes therapeutic potency of Islet-like cells. METHODS: Rat MSCs were isolated from Wistar rats and cultured. Passaged MSCs were induced to differentiate into islet-like cells under following conditions: pre-induction with L-DMEM including 10 mmol/L nicotinamide+l mmol/L β-mercaptoethanol+200 mL/L fetal calf serum (FSC) for 24 h, followed by induction with serum free H-DMEM solution including 10 mmol/L nicotinamide+1 mmol/L,β-mercaptoethanol for 10 h. Differentiated cells were observed under inverse microscopy, insulin and nestin expressed in differentiated cells were detected with immunocytochemistry. Insulin excreted from differentiated cells was tested with radioimmunoassay. Rat diabetic models were made to test in vivo function of differentiated MSCs. RESULTS: Typical islet-like clustered cells were observed. Insulin mRNA and protein expressions were positive in differentiated cells, and nestin could be detected in predifferentiated cells. Insulin excreted from differentiated MSCs (446.93±102.28 IU/L) was much higher than that from pre-differentiated MSCs (2.45+0.81 IU/L (P<0.01). Injected differentiated MSCs cells could down-regulate glucose level in diabetic rats. CONCLUSION: Islet-like functional cells can be differentiated from marrow mesenchymal stem cells, which may be a new procedure for clinical diabetes stem -cell therapy, these cells can control blood glucose level in diabetic rats. MSCs may play an important role in diabetes therapy by islet differentiation and transplantation.展开更多
目的:体外分离纯化人牙周膜干细胞,研究其生物学特性及表型特点并进行初步鉴定。方法:采用胶原酶和D ispase酶联合消化法培养人牙周膜干细胞,有限稀释法分离纯化,将克隆形成的细胞通过透射电镜观察其超微结构、流式细胞仪细胞周期分析...目的:体外分离纯化人牙周膜干细胞,研究其生物学特性及表型特点并进行初步鉴定。方法:采用胶原酶和D ispase酶联合消化法培养人牙周膜干细胞,有限稀释法分离纯化,将克隆形成的细胞通过透射电镜观察其超微结构、流式细胞仪细胞周期分析及免疫组织化学染色技术检测抗波形丝蛋白(V im en-tin)、STRO-1、骨粘素(ON)、骨涎蛋白(BSP)的表达。结果:获得纯化人牙周膜干细胞,在透射电镜下,这种细胞核质比例大,核大,细胞器少;流式细胞仪细胞周期分析大多数细胞处于G0/G1期,为慢周期性;该细胞抗波形丝蛋白、STRO-1表达阳性、骨粘连素、骨桥蛋白弱阳性表达。结论:提供了比较有效的分离纯化牙周膜干细胞的方法。该方法分离的人牙周膜干细胞具有干细胞的超微结构、细胞周期及表型特点。展开更多
基金Supported by the National Natural Science Foundation of China,No.30170911
文摘AIM: To explore the possibility of marrow mesenchymal stem cells (MSC) in vitro differentiating into functional isletlike cells and to test the diabetes therapeutic potency of Islet-like cells. METHODS: Rat MSCs were isolated from Wistar rats and cultured. Passaged MSCs were induced to differentiate into islet-like cells under following conditions: pre-induction with L-DMEM including 10 mmol/L nicotinamide+l mmol/L β-mercaptoethanol+200 mL/L fetal calf serum (FSC) for 24 h, followed by induction with serum free H-DMEM solution including 10 mmol/L nicotinamide+1 mmol/L,β-mercaptoethanol for 10 h. Differentiated cells were observed under inverse microscopy, insulin and nestin expressed in differentiated cells were detected with immunocytochemistry. Insulin excreted from differentiated cells was tested with radioimmunoassay. Rat diabetic models were made to test in vivo function of differentiated MSCs. RESULTS: Typical islet-like clustered cells were observed. Insulin mRNA and protein expressions were positive in differentiated cells, and nestin could be detected in predifferentiated cells. Insulin excreted from differentiated MSCs (446.93±102.28 IU/L) was much higher than that from pre-differentiated MSCs (2.45+0.81 IU/L (P<0.01). Injected differentiated MSCs cells could down-regulate glucose level in diabetic rats. CONCLUSION: Islet-like functional cells can be differentiated from marrow mesenchymal stem cells, which may be a new procedure for clinical diabetes stem -cell therapy, these cells can control blood glucose level in diabetic rats. MSCs may play an important role in diabetes therapy by islet differentiation and transplantation.
文摘目的:体外分离纯化人牙周膜干细胞,研究其生物学特性及表型特点并进行初步鉴定。方法:采用胶原酶和D ispase酶联合消化法培养人牙周膜干细胞,有限稀释法分离纯化,将克隆形成的细胞通过透射电镜观察其超微结构、流式细胞仪细胞周期分析及免疫组织化学染色技术检测抗波形丝蛋白(V im en-tin)、STRO-1、骨粘素(ON)、骨涎蛋白(BSP)的表达。结果:获得纯化人牙周膜干细胞,在透射电镜下,这种细胞核质比例大,核大,细胞器少;流式细胞仪细胞周期分析大多数细胞处于G0/G1期,为慢周期性;该细胞抗波形丝蛋白、STRO-1表达阳性、骨粘连素、骨桥蛋白弱阳性表达。结论:提供了比较有效的分离纯化牙周膜干细胞的方法。该方法分离的人牙周膜干细胞具有干细胞的超微结构、细胞周期及表型特点。
