Three-Dimensional Culture Promotes the Differentiation of Human Dental Pulp Mesenchymal Stem Cells Into Insulin-Producing Cells for Improving the Diabetes Therapy.

Bingbing Xu,Bo Niu,Daoyang Fan,Zhen Guan,Jianhua Wang,Jing Li,Zhendong Wang,Yunshan Zhao,Xiuwei Wang

doi:10.3389/fphar.2019.01576

Abstract

IntroductionDiabetes is a metabolic disease with a high incidence and serious harm to human health. Islet β-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.

Highlights

Diabetes is a metabolic disease with a high incidence and serious harm to human health
Morphology and Multilineage Differentiation Potential human dental pulp stem cells (hDPSCs) exhibited typical mesenchymal stem cells (MSCs) morphology, and shared the similar fibroblast-like, spindle shape and expressed high capacity to adhere to plastic culture flasks (Figure 1A)
The morphology of hDPSCs showed typical morphology of MSCs, which was relatively uniform in morphology, similar to the fusiform shape of fibroblasts, and showed high adhesion to plastic culture flasks (Figure 1A)

Summary

Introduction

Diabetes is a metabolic disease with a high incidence and serious harm to human health. The main treatments for diabetes include diet control, regular blood glucose testing, oral hypoglycemic chemotherapy and insulin supplementation. Such treatment methods can not reconstruct the physiological blood sugar regulation function of the body, and can not avoid the occurrence of severe hypoglycemia and long-term complications (Nathan, 1993). Another treatment method is to replace endogenous b cells by islet transplantation. Islet transplantation has been successfully used in clinical practice, the limited supply of islets and the side effects of immunosuppressive therapy have largely hampered the widespread use of this therapy (Porat and Dor, 2007), prompting us to search for alternative sources of islet cells

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