Composite Scaffolds of Decellularized Placenta/PRP Enhance the Differentiation of Adipose Tissue-Derived Mesenchymal Stem Cells Into Insulin-Producing Cells In Vitro.

Abstract

In the current in vitro experiment, we fabricated and characterized placenta/platelet-rich plasma (PL/Pt) composite scaffolds and evaluated their effect on differentiating adipose stem cells (ASCs) into insulin-producing cells (IPCs) in vitro. The human placenta (PL) was decellularized (dPL), characterized, and digested in pepsin. PRP was extracted using a two-step centrifugation process and then freeze-dried. PL/Pt composite scaffolds were fabricated from PL with various PRP concentrations (1, 2.5, and 5 mg·mL) and characterized. ASCs were isolated from adipose tissue and characterized using flow cytometry and multilineage differentiation assays. ASCs were seeded onto PL/Pt scaffolds and differentiated into IPCs. The decellularized placenta retained its collagen content and had minimal cellular and DNA content. ASCs expressed CD73, CD90, and CD105 but not CD34 and CD45 and differentiated into osteoblasts and adipocytes. PL/Pt scaffolds showed appropriate morphological and chemical properties with improved porosity, swelling, and degradation rates. Seeding cells on the PL scaffolds increased PDX1, GLUT2, and insulin expression significantly compared to cells cultured in plates. Cells seeded on PL/Pt2.5 and PL/Pt5 scaffolds showed a remarkable increase in PDX1, GLUT2, and insulin expression. Additionally, differentiated cells cultivated on PL/Pt scaffolds exhibited enhanced insulin and C-peptide secretion in response to variations in glucose levels. PL/Pt composite scaffolds provide a biomimetic microenvironment supporting ASCs' survival while enhancing their differentiation into IPCs. This approach could be a promising strategy for replacing the damaged β cell population in diabetic patients.