Study of mouse adipose tissue-derived mesenchymal stem cells differentiation to hepatocyte-like using gelatin/laminin 3D scaffold

Using 3D culture to increase cell density and creating a suitable bedrock for the growth and proliferation and differentiation of stem cells is one of the important methods for their application in tissue engineering. In this study, mesenchymal stem cells of adipose tissue because of ease of access and abundance have been used to differentiate into hepatocyte-like cells to investigate the effect of 3D cultures on gelatin/laminin 3D scaffolds.

Mesenchymal stem cells were extracted from subcutaneous fat of the abdominal region of C57 mice by enzymatic digestion using collagenase enzyme. The extracted stem cells were confirmed by flow cytometry analysis and observation of the expression of specific stem cells markers such as CD105, CD44, and the absence of specific leukocyte (CD34) and hematopoietic (CD45). The non-toxicity of the scaffold was also investigated by the MTT method. Stem cells were cultured on a polystyrene surface and laminin and gelatin/laminin scaffold and treated with growth factors for 21 days in two stages. Cell differentiation was investigated by biochemical methods including examination of secretion indices such as urea and glycogen storage.

The expression of specific stem cells markers such as CD105, CD44, and the absence of CD34 and CD45 were proved. The ability to differentiate these cells into adipocytes and osteocytes has been proven.

The results show the presence of a three-dimensional gelatin/laminin scaffold increases the adhesion, proliferation, and differentiation of fatty mesenchymal cells to hepatocyte-like cells.