Three-Dimensional Culture Promotes the Differentiation of Human Dental Pulp Mesenchymal Stem Cells Into Insulin-Producing Cells for Improving the Diabetes Therapy
Three-Dimensional Culture Promotes the Differentiation of Human Dental Pulp Mesenchymal Stem Cells Into Insulin-Producing Cells for Improving the Diabetes Therapy
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三维培养促进人牙髓间充质干细胞分化为胰岛素产生细胞，改善糖尿病治疗
DOI：
10.3389/fphar.2019.01576
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发表时间：
2020-01-24
影响因子：
5.6
通讯作者：
Niu, Bo
中科院分区：
文献类型：
--
作者：
Xu, Bingbing;Fan, Daoyang;Niu, Bo
引言糖尿病是一种发病率高、严重危害人类健康的代谢性疾病。胰岛β细胞功能缺陷可发生在1型糖尿病和2型糖尿病的晚期。研究表明，干细胞是生物工程再生医学中一种有前途的新方法。在干细胞分化的研究中，三维（3D）细胞培养比二维（2D）细胞培养更能模拟体内细胞生长的微环境。细胞与细胞、细胞与细胞外基质之间的自然接触可以调节发育过程，促进人工再生器官和组织的形成。 IV型、VI型胶原蛋白和层粘连蛋白是胰岛中最丰富的细胞外基质成分。 Matrigel，一种富含层粘连蛋白和 IV 型胶原的基底膜基质生物材料。材料与方法我们利用Matrigel生物材料物理包埋人牙髓干细胞（hDPSCs），为细胞提供载体和3D培养条件，探索并比较hDPSCs在2D或3D培养条件下分化为胰岛素生成细胞（IPCs）的制备方法和初步机制。我们首先通过模拟体内胰腺生成的关键事件来设计和筛选策略，成功建立了从hDPSCs获得IPCs的新方法。采用Activin A、Noggin和小分子化合物协同诱导hDPSCs在2D培养条件下逐步分化为定形内胚层样细胞、胰腺祖细胞样细胞和IPCs。然后，我们使用Matrigel模拟体内微环境，在Matrigel中诱导hDPSCs分化为IPCs，评估并比较2D和3D培养条件之间的效率。结果结果显示，生长因子和小分子化合物的协同组合以及3D培养促进了hDPSCs向IPCs的分化，显着增强了IPCs胰岛素和C肽的释放。讨论为获得大量功能性IPC用于再生医学中的疾病建模和最终细胞治疗提供了重要支持。
IntroductionDiabetes is a metabolic disease with a high incidence and serious harm to human health. Islet beta-cell function defects can occur in the late stage of type 1 diabetes and type 2 diabetes. Studies have shown that stem cell is a promising new approach in bioengineering regenerative medicine. In the study of stem cell differentiation, three-dimensional (3D) cell culture is more capable of mimicking the microenvironment of cell growth in vivo than two-dimensional (2D) cell culture. The natural contact between cells and cells, and cells and extracellular matrix can regulate the development process and promote the formation of the artificial regenerative organs and organization. Type IV, VI collagen and laminin are the most abundant extracellular matrix components in islets. Matrigel, a basement membrane matrix biomaterial rich in laminin and collagen IV. Materials and MethodsWe used Matrigel biomaterial to physically embed human dental pulp stem cells (hDPSCs) to provide vector and 3D culture conditions for cells, and we explored and compared the preparation methods and preliminary mechanisms of differentiation of hDPSCs into insulin-producing cells (IPCs) under 2D or 3D culture conditions.We first designed and screened the strategy by mimicking the critical events of pancreatogenesis in vivo, and succeeded in establishing a new method for obtaining IPCs from hDPSCs. Activin A, Noggin, and small molecule compounds were used to synergistically induce hDPSCs to differentiate into definitive endoderm-like cells, pancreatic progenitor like cells and IPCs step by step under 2D culture conditions. Then, we used Matrigel to simulate the microenvironment in vivo, induced hDPSCs to differentiate into IPCs in Matrigel, evaluated and compared the efficiency between 2D and 3D culture conditions. ResultsThe results showed that the synergistic combination of growth factors and small molecule compounds and 3D culture promoted the differentiation of hDPSCs into IPCs, significantly enhancing the release of insulin and C-peptide from IPCs. DiscussionSignificant support is provided for obtaining a large number of functional IPCs for disease modeling and final cell therapy in regenerative medicine.