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目的探讨回输至链脲佐菌素(STZ)诱导的糖尿病小鼠体内的骨髓干细胞能否在体内被诱导分化为胰岛素阳性细胞。方法将实验小鼠分为3组,即正常组、对照组(糖尿病组)和实验组(糖尿病并接受骨髓干细胞回输组)。待回输的骨髓干细胞取自3周龄雄性Balb/c小鼠,按改良的Verfaillie实验室的培养条件分离培养骨髓干细胞。STZ诱导的6~8周龄雌性Balb/c糖尿病小鼠为回输骨髓干细胞的受者。实验组小鼠尾静脉注射1~2×106CFSE标记的骨髓干细胞。观察各组小鼠血糖、体重变化。4周后实验结束时,取胰腺行Y染色体PCR扩增、胰岛素免疫荧光和CFSE检测,了解骨髓干细胞在胰腺的定位及诱导分化情况;计算、比较3组胰岛素免疫组化切片中胰腺组织胰岛面积和胰岛素阳性面积,了解体内回输骨髓干细胞能否诱导分化为胰岛素阳性细胞。结果实验过程中,实验组和对照组小鼠的血糖无明显下降,体重无明显增加,而正常组血糖保持在正常范围,体重逐渐增加;实验组中,胰腺组织Y染色体(n=6)的检出比例为6/6(100%);实验组小鼠胰腺组织有CFSE标记细胞检出,每视野下胰岛素染色阳性同时CFSE染色阳性细胞数为1.2±1.1;胰岛面积,实验组为(10172±6303)μm2,对照组为(9857±4444)μm2,正常组为(17175±10495)μm2。实验组与对照组比较,差异无统计学意义;与正常组比较,实验组和对照组显著小于正常组(均P<0.05);胰岛素染色面积,实验组为(474±380)μm2,对照组为(329±499)μm2,正常组为(1527±1788)μm2;实验组大于对照组(P<0.05),小于正常组(P<0.05)。结论在STZ诱导的糖尿病小鼠回输骨髓干细胞,有少部分可定位于胰腺,具备胰岛素阳性细胞的特征。
Objective To investigate whether bone marrow stem cells reintroduced into streptozotocin (STZ) -induced diabetic mice can be induced to differentiate into insulin-positive cells in vivo. Methods The experimental mice were divided into 3 groups: normal group, control group (diabetic group) and experimental group (diabetes mellitus receiving bone marrow stem cell transfusion group). Bone marrow stem cells to be transfused were taken from 3-week-old male Balb / c mice and bone marrow stem cells were isolated and cultured according to the modified Verfaillie laboratory conditions. STZ-induced 6-8 week old female Balb / c diabetic mice were recipients of transfused bone marrow stem cells. The experimental mice were injected with 1 ~ 2 × 106CFSE labeled bone marrow stem cells through tail vein. The blood glucose and body weight of mice in each group were observed. At the end of the experiment at the end of 4 weeks, pancreas was obtained by PCR amplification of Y chromosome, immunofluorescence of insulin and detection of CFSE to understand the localization and differentiation of bone marrow stem cells in the pancreas. Comparisons of pancreatic islet area And insulin-positive area, understand the in vivo return of bone marrow stem cells can differentiate into insulin-positive cells. Results During the experiment, there was no significant decrease in blood glucose and no increase in body weight in the experimental group and control group, while the normal group maintained normal blood glucose level and increased body weight. In the experimental group, the Y chromosome (n = 6) The detection rate was 6/6 (100%). The CFSE-labeled cells were detected in the pancreas of the experimental group. The numbers of CFSE-positive cells in every field were 1.2 ± 1.1. The islet area in the experimental group was (10172) ± 6303) μm2 in the control group and (9857 ± 4444) μm2 in the control group and 17175 ± 10495 μm2 in the normal group. Compared with the normal group, the experimental group and the control group were significantly smaller than the normal group (all P <0.05); the area of insulin staining was (474 ± 380) μm2 in the experimental group and the control group (329 ± 499) μm2 in the normal group and (1527 ± 1788) μm2 in the normal group. The experimental group was larger than the control group (P <0.05) and smaller than the normal group (P <0.05). Conclusions A small number of bone marrow stem cells were transfused from STZ-induced diabetic mice and localized to the pancreas with the characteristics of insulin-positive cells.