Isolation of tissue progenitor cells from duct-ligated salivary glands of swine
Isolation of tissue progenitor cells from duct-ligated salivary glands of swine
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DOI：
10.1089/clo.2006.0022
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发表时间：
2007-03-01
期刊：
影响因子：
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通讯作者：
Endo, Fumio
中科院分区：
文献类型：
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作者：
Matsumoto, Shirou;Okumura, Kenji;Endo, Fumio
组织干细胞参与损伤后组织的再生。组织损伤刺激正常静止的组织干细胞分化和增殖，在替换和/或修复受损细胞的过程中，从而影响组织再生。唾液腺保留了频繁再生的能力。先前，我们从小鼠和大鼠的受损唾液腺中分离出分化为肝脏和胰腺谱系的祖细胞。分离的祖细胞cd49f阳性，细胞内层粘连蛋白阳性，在I型胶原上增殖，同时保持其多能性。本研究对结扎猪唾液腺主排泄管诱导的组织干细胞进行了分析。腺导管结扎后，腺泡细胞因凋亡而萎缩，上皮细胞随后增殖。我们培养了从导管结扎的唾液腺中获得的细胞，并通过有限的稀释纯化了细胞。分离的细胞CD29、CD49f、细胞内层粘连蛋白、AFP、CK19、CK18和Thy-1（CD90）呈阳性，c-Kit （CD117）呈弱阳性。三维形成后，细胞表达胰岛素和白蛋白。我们将这些细胞命名为猪唾液腺源性祖细胞。在球形培养中，在烟酰胺和/或GLP-1处理的分化培养基中，胰岛素和白蛋白的基因表达显著增加（5倍），胰岛素的基因表达也显著增加（3.8倍）。白蛋白和胰岛素的表达分别是猪肝细胞和胰腺内分泌细胞的1/10和1/4倍。分化后的SGP细胞在葡萄糖和钾的刺激下可释放胰岛素。这些结果表明，猪SGP细胞在功能上可以分化为肝细胞和β细胞。猪SGP细胞是大型动物内胚层再生模型治疗和分析的有用工具。
Tissue stem cells participate in the repopulation of tissue after injury. Tissue injury stimulates the normally quiescent tissue stem cells to differentiate and proliferate, in the process of replacing and/or repairing the damaged cells, and hence effecting tissue regeneration. The salivary glands retain the ability for frequent regeneration. Previously, we isolated progenitor cells from the injured salivary glands of mice and rats that differentiated into hepatic and pancreatic lineages. The isolated progenitors were CD49f-positive and intracellular laminin-positive, and proliferated on type I collagen while maintaining their multipotency. In this study, we analyzed the tissue stem cells induced by ligating the main excretory duct of the salivary gland in swine. After duct ligation of the gland, acinar cells receded due to apoptosis, and epithelial cells subsequently proliferated. We cultured cells obtained from the duct-ligated salivary gland and purified the cells by limited dilution. The isolated cells were positive for CD29, CD49f, intracellular laminin, AFP, CK19, CK18, and Thy-1(CD90), and weakly positive for c-Kit (CD117). After three-dimensional formation, the cells expressed insulin and albumin. We designated the cells as swine salivary gland-derived progenitor cells. Gene expression of insulin and albumin was significantly increased (five-fold) and that of insulin was also increased (3.8-fold) with differentiation medium with nicotinamide and/or GLP-1 treatment in spherical culture. The expressions of albumin and insulin were 1/10-fold and 1/4-fold compared to porcine hepatocytes and pancreatic endocrine cells. The differentiated SGP cells could release insulin, which were stimulated by glucose and potassium. These results indicate that swine SGP cells could differentiate into hepatocytes and beta-cells, functionally. Swine SGP cells were useful tools for therapy and analyzing endodermal regenerative models in large animals.