Isolation and characterization of human islet stellate cells
Isolation and characterization of human islet stellate cells
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人胰岛星状细胞的分离和表征
DOI：
10.1016/j.yexcr.2015.11.002
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发表时间：
2016
影响因子：
3.7
通讯作者：
Sun Zilin
中科院分区：
文献类型：
--
作者：
Zha Min;Xu Wei;Jones Peter M.;Sun Zilin
背景与目的我们已经证明，可以从大鼠胰岛中分离出与经典胰星状细胞(PSCs)表型相似的胰岛星状细胞(ISCs)，它们可能参与了2型糖尿病(T2 DM)的胰岛纤维化。本研究旨在确定人的胰岛是否也含有干细胞。材料和方法采用标准的组织块技术，从新鲜分离的人胰岛中获取丰富的人胰岛素样干细胞。采用免疫荧光方法检测α-平滑肌肌动蛋白(α-SMA)、结蛋白、波形蛋白、胶质纤维酸性蛋白(GFAP)对人间质干细胞的影响。采用细胞计数试剂盒-8(CCK-8)检测细胞增殖情况。采用创面愈合实验和跨孔迁移实验评价ISC的迁移能力。I型胶原(COL-I)、III型胶原(COL-III)和纤维连接蛋白(FN)免疫荧光染色鉴定ISC合成的细胞外基质(ECM)。成脂和成骨分化检测干细胞潜能。结果在培养条件下，人胰岛细胞分化为三角形的ISC。传代后的间质细胞表达α-SMA、结蛋白、波形蛋白、胶质原纤维酸性蛋白，表达Col-I、Col-III和Fn。ISC的增殖和迁移能力明显慢于PSC。人PSC和ISC在体外均能分化为脂肪样细胞和成骨细胞样细胞。结论与我们之前的大鼠实验相似，本研究发现人胰岛也含有ISC，其表型与人PSC相似，但不完全相同。
Background and aimsWe have previously demonstrated that islet stellate cells (ISCs) exhibiting a similar phenotype to classical pancreatic stellate cells (PSCs) could be isolated from rat islets, where they may contribute to islet fibrosis in type 2 diabetes mellitus (T2DM). This study was designed to determine whether human islets also contain ISC.Materials and methodsUsing standard explants techniques, human ISCs were enriched from freshly isolated human islets. Immunofluorescence visualization of markers for PSCs(α-smooth muscle actin;α-SMA), desmin, vimentin, glial fibrillary acidic protein (GFAP) was used to characterize the human ISC. Cell counting kit-8 (CCK-8) was used to assess the proliferation of ISC. The wound-healing assay and the transwell migration were used to assess the migration capacity of ISC. Immunofluorescence against collagen typesI (col-I), collagen typesIII (col-III) and fibronectin (FN) was performed to identify extracellular matrix (ECM) component synthesized by ISC. Adipogenic and osteogenic differentiation were tried to detected stem cell potential.ResultsIn culture, ISC with triangular shape grow out from human islets. The passaged ISC expressed α-SMA, desmin, vimentin, GFAP and was positive for col-I, col-III and FN. The proliferation and migration ability of ISC was significantly slower than those of PSC. And both the human PSC and ISC were able to differentiate in vitro into adipocyte- and osteoblast-like cells.ConclusionSimilar to our previous rat experiment, the current study shows that human islets also contain ISC which is phenotypically similar but not identical to human PSC.