Gao Fan, Wu Yangnan, Wen Hebao, Zhu Wanwan, Ren Han, Guan Weijun, Tian Xiuzhi
College of Human Movement Science, Harbin Sport University, Harbin, Heilongjiang, 150040, China.
Institute of Animal Science of CAAS, Beijing 100193, China.
Tissue Cell. 2019 Feb;56:60-70. doi: 10.1016/j.tice.2018.12.001. Epub 2018 Dec 6.
Stem cells are most likely to solve all three of diabetes's problems at once, but the previous studies have mostly focused on bone marrow mesenchymal stem cells (MSCs) and adipose tissue-derived MSCs, and few studies have been done on pancreatic MSCs. In this study, pancreatic was collected to isolate MSCs from bovine, and then their biological characteristics such as growth kinetics, surface antigen, and multilineage potential were examined. Pancreatic MSCs of bovine (B-PMSCs) could be cultured for 65 passages in vitro. Growth kinetics analyses indicated that B-PMSCs had a strong capacity for self-renewal in vitro and their proliferation capacity appeared to decrease by passaging. Surface antigen detection showed that B-PMSCs expressed CD29, CD44, CD73, CD90, CD106, CD166, Vimentin, Nestin and Insulin, but not expressed CD34 and CD45. Furthermore, B-PMSCs could be induced to differentiate into adipocytes, osteoblasts and smooth muscle cells as indicated by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence. Most importantly, insulin-secreting cell differentiation of B-PMSCs exhibited islet-like clusters and dithizone staining displayed scarlet, and the response of the islet-like clusters to glucose suggested that high concentration glucose (20 mM) could quickly and persistently stimulate insulin release, and from the 2.0 h of the stimulation, the insulin of 20 mM glucose group were significantly higher than the 5.5 mM group. The B-PMSCs were isolated successfully, and the cells owned powerful self-renewal ability and multiple differentiative potential. Therefore, the present study plays an important role by providing a PMSCs choice for cell therapy of diabetes and tissue engineering.
干细胞最有可能一次性解决糖尿病的所有三个问题，但先前的研究大多集中在骨髓间充质干细胞（MSCs）和脂肪组织来源的间充质干细胞上，而对胰腺间充质干细胞的研究较少。在本研究中，收集牛胰腺以分离间充质干细胞，然后检测其生长动力学、表面抗原和多向分化潜能等生物学特性。牛胰腺间充质干细胞（B-PMSCs）可在体外培养65代。生长动力学分析表明，B-PMSCs在体外具有很强的自我更新能力，其增殖能力似乎会随着传代而降低。表面抗原检测显示，B-PMSCs表达CD29、CD44、CD73、CD90、CD106、CD166、波形蛋白、巢蛋白和胰岛素，但不表达CD34和CD45。此外，逆转录-聚合酶链反应（RT-PCR）和免疫荧光显示，B-PMSCs可被诱导分化为脂肪细胞、成骨细胞和平滑肌细胞。最重要的是，B-PMSCs向胰岛素分泌细胞的分化表现为胰岛样簇，双硫腙染色呈猩红色，胰岛样簇对葡萄糖的反应表明，高浓度葡萄糖（20 mM）可快速持续刺激胰岛素释放，从刺激2.0小时起，20 mM葡萄糖组的胰岛素水平显著高于5.5 mM组。成功分离出B-PMSCs，这些细胞具有强大的自我更新能力和多向分化潜能。因此，本研究为糖尿病细胞治疗和组织工程提供了一种胰腺间充质干细胞选择，具有重要意义。
