Decellularized and solubilized pancreatic stroma promotes the in vitro proliferation, migration and differentiation of BMSCs into IPCs
Decellularized and solubilized pancreatic stroma promotes the in vitro proliferation, migration and differentiation of BMSCs into IPCs
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脱细胞和溶解的胰腺基质促进BMSCs体外增殖、迁移和分化为IPCs
DOI：
10.1007/s10561-019-09777-4
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发表时间：
2019-09-01
影响因子：
1.5
通讯作者：
Wang, Zhiwei
中科院分区：
文献类型：
--
作者：
Guo, Yibing;Chen, Saisai;Wang, Zhiwei
骨髓间充质干细胞(BMSCs)具有向胰岛素分泌细胞(IPC)分化的能力。脱细胞器官衍生的生物支架可作为种子细胞的载体，在再生医学中具有广泛的应用前景。本研究观察从去细胞胰腺获取的天然胰腺基质对骨髓间充质干细胞增殖、迁移和向内皮祖细胞分化的影响，并探讨其潜在的分子机制。以Triton X-100/氢氧化铵为灌注剂，经胃酶和盐酸混合消化法制备基质溶液，得到脱细胞胰腺生物支架。采用三步诱导法将骨髓间充质干细胞分化为胰岛样细胞。通过细胞形态观察、胰岛素释放试验、qRT-PCR分析和免疫印迹分析，评价有无基质对细胞生物学行为的影响。结果表明，去细胞胰腺来源的基质能促进BMSCs的增殖和迁移。此外，还能促进与内源性胰岛形态相似的IPC的形成。在诱导分化过程中，间质的存在显著增加了胰岛素1、2和PDX-1的表达以及胰岛素的释放。伴随而来的是第三阶段细胞团中Akt和ERK的磷酸化增加，这可分别被抑制剂PD98059和LY294002阻止。综上所述，脱细胞的胰腺间质可以促进BMSCs的增殖、迁移和向IPC分化，这涉及Akt和ERK信号通路的激活。
Bone marrow-derived mesenchymal stem cells (BMSCs) have the ability to differentiate into insulin-producing cells (IPCs). Bio-scaffolds derived from decellularized organs can act as a carrier for seed cells and may have broad applications in regenerative medicine. This study investigated the effect of native pancreatic stroma obtained from decellularized pancreas on the proliferation, migration and differentiation of BMSCs into IPCs, and explored the potential underlying molecular mechanism. The decellularized pancreas bio-scaffold was obtained by perfusion with Triton X-100/ammonium hydroxide, followed by digestion with a mixture of pepsin and hydrochloric acid to prepare the stroma solution. Islet-like cells were differentiated from BMSCs by a three-step induction method. The differences on the cytological behavior with or without stroma were evaluated by morphological observation, insulin release assay, qRT-PCR assay and western blot analysis. Our results showed that, stroma derived from decellularized pancreas could promote the proliferation and migration of BMSCs. Furthermore, the formation of IPCs could also be promoted, which possessed similar morphology to endogenous islets. During the induced differentiation process, the presence of stroma significantly increased the expression of insulin 1, insulin 2 and Pdx-1, as well as insulin release. This was accompanied by an increase in the phosphorylation of Akt and ERK in third stage cell clusters, which was prevented by the addition of the inhibitors PD98059 and LY294002, respectively. In summary, decellularized pancreatic stroma could promote the proliferation, migration and differentiation of BMSCs into IPCs, and this involved the activation of Akt and ERK signal pathways.