Dorisetty Raj-K, Kiran Sashi-G, Umrani Malathi-R, Boindala Sesikeran, Bhonde Ramesh-R, Venkatesan Vijayalakshmi
Department of Biochemistry, National Institute of Nutrition, Tarnaka, Hyderabad 500007, India.
World J Gastroenterol. 2008 Dec 14;14(46):7112-6. doi: 10.3748/wjg.14.7112.
To localize nestin positive cells (NPC) in pancreatic tissue of mice of different ages.
Paraffin sections of 6-8 mum of fixed pancreatic samples were mounted on poly-L-lysine coated slides and used for Immunolocalization using appropriate primary antibodies (Nestin, Insulin, Glucagon), followed by addition of a fluorescently labeled secondary antibody. The antigen-antibody localization was captured using a confocal microscope (Leica SP 5 series).
In 3-6 d pups, the NPC were localized towards the periphery of the endocrine portion, as evident from immunolocalization of insulin and glucagon, while NPC were absent in the acinar portion. At 2 wk, NPC were localized in both the exocrine and endocrine portions. Interestingly, in 4-wk-old mice NPC were seen only in the endocrine portion, towards the periphery, and were colocalised with the glucagon positive cells. In the pancreas of 8- wk-old mice, the NPC were predominantly localized in the central region of the islet clusters, where immunostaining for insulin was at a maximum.
We report for the first time the immunolocalization of NPC in the pancreas of mice of different ages (3 d to 8 wk) with reference to insulin and glucagon positive cells. The heterogeneous localization of the NPC observed may be of functional and developmental significance and suggest(s) that mice pancreatic tissue can be a potential source of progenitor cells. NPC from the pancreas can be isolated, proliferated and programmed to differentiate into insulin secreting cells under the appropriate microenvironment.
在不同年龄小鼠的胰腺组织中定位巢蛋白阳性细胞(NPC)。
将固定的胰腺样本制成6 - 8微米的石蜡切片,置于经聚-L-赖氨酸包被的载玻片上,使用适当的一抗(巢蛋白、胰岛素、胰高血糖素)进行免疫定位,随后添加荧光标记的二抗。使用共聚焦显微镜(徕卡SP 5系列)捕捉抗原-抗体定位。
在3 - 6日龄幼鼠中,从胰岛素和胰高血糖素的免疫定位可见,NPC定位于内分泌部分的周边,而腺泡部分不存在NPC。在2周龄时,NPC定位于外分泌和内分泌部分。有趣的是,在4周龄小鼠中,NPC仅在内分泌部分的周边被观察到,并与胰高血糖素阳性细胞共定位。在8周龄小鼠的胰腺中,NPC主要定位于胰岛簇的中央区域,此处胰岛素免疫染色最强。
我们首次报道了不同年龄(3日龄至8周龄)小鼠胰腺中NPC相对于胰岛素和胰高血糖素阳性细胞的免疫定位。观察到的NPC的异质性定位可能具有功能和发育意义,并提示小鼠胰腺组织可能是祖细胞的潜在来源。胰腺中的NPC可以在适当的微环境下被分离、增殖并编程分化为胰岛素分泌细胞。