3D Adipose-Derived Stem Cells to Hepatocyte-Like Cell Scaffold to Investigate the Drug-Induced Hepatotoxicity.

Abstract

In this article, the three-dimensional (3D) adipose-derived stem cells (ADSCs) differentiated into hepatocyte-like cells were fabricated on polycaprolactone (PCL)/sodium alginate/gelatin hydrogel scaffolds. To this end, the 3D scaffold was fabricated applying PCL, sodium alginate, and gelatin hydrogel using electrospinning machine. During the fabrication and testing of scaffold, the human ADSCs were transformed into hepatocyte cells using the mesenchymal process for 21 days. Human placenta extract (HPE) was used in the cell culture to increase cell growth, proliferation, and differentiation. The results indicated the tensile elastic modulus of the scaffold at 85.5 ± 15.5 kPa. The contact angle and porosity values were 67.7° and 71.351%, respectively. The swelling and degradation percentages were 22.308%, 1.778% (1 h), 53.385%, 5.111% (24 h), and finally 71.318% and 17.778% (14 days). Based on the dynamic light scattering image, the diameter size of the HPE and alginate nanoparticles was between 10 and 100 nm. According to the cell viability analysis for 72 h, the optimized HPE value was 0.64%. Based on the results of the oil-red O staining and alizarin red staining, ∼80% of the ADSCs differentiated successfully into adipogenic and osteogenically formats. The orders of different formats of ADSCs for cell proliferations, resistance to cisplatin drug experiments, fold change factor for PXR, CYP1A2, and albumin gene concentration were 3D ADSC plus HPE, 2D ADSC plus HPE, 3D ADSC, and ADSC models as the first to fourth, respectively. Due to flow cytometry test, surface markers CD31 and CD45 were negative, whereas CD90 and CD105 were positive.