The enhancement of differentiating adipose derived mesenchymal stem cells toward hepatocyte like cells using gelatin cryogel scaffold

• Published in: Biochemical and biophysical research communications Vol. 491; no. 4; pp. 1000 - 1006
• Main Authors: Ghaderi Gandomani, Maryam, Sahebghadam Lotfi, Abbas, Kordi Tamandani, Dormohammad, Arjmand, Sareh, Alizadeh, Shaban
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 30-09-2017
• Subjects: Adipose Tissue - cytology; Adult; Cell Differentiation; Cells, Cultured; Cryogel scaffold; Cryogels - chemistry; Female; Flow Cytometry; Gelatin; Gelatin - chemistry; Hepatic differentiation; Hepatocytes - cytology; Humans; Liver tissue engineering; Mesenchymal stem cell; Mesenchymal Stromal Cells - cytology; Tissue Scaffolds - chemistry; Mesenchymal stem cell; PAS; Cryogel scaffold; AFP; MTT; ECM; ACTB; Liver tissue engineering; hADSCs; Hepatic differentiation; Gelatin; TCPS; ALB; SEM; MSC; CK-18; CK-19
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2017.07.167

Liver tissue engineering creates a promising methodology for developing functional tissue to restore or improve the function of lost or damaged liver by using appropriate cells and biologically compatible scaffolds. The present paper aims to study the hepatogenic potential of human adipose derived mesenchymal stem cells (hADSCs) on a 3D gelatin scaffold in vitro. For this purpose, mesenchymal stem cells were isolated from human adipose tissue and characterized by flowcytometry analysis and mesodermal lineage differentiation capacity. Then, porous cryogel scaffolds were fabricated by cryogelating the gelatin using glutaraldehyde as the crosslinking agent. The structure of the scaffolds as well as the adhesion and proliferation of the cells were then determined by Scanning Electron Microscopy (SEM) analysis and MTT assay, respectively. The efficiency of hepatic differentiation of hADSCs on 2D and 3D culture systems has been assessed by means of morphological, cytological, molecular and biochemical approaches.
Based on the results of flowcytometry, the isolated cells were positive for hMSC specific markers and negative for hematopoietic markers. Further, the multipotency of these cells was confirmed by adipogenic and osteogenic differentiation and the highly porous structure of scaffolds was characterized by SEM images. Biocompatibility was observed in the fabricated gelatin scaffolds and the adhesion and proliferation of hADSCs were promoted without any cytotoxicity effects. In addition, compared to 2D TCPS, the fabricated scaffolds provided more appropriate microenvironment resulting in promoting the differentiation of hADSCs toward hepatocyte-like cells with higher expression of hepatocyte-specific markers and appropriate functional characteristics such as increased levels of urea biosynthesis and glycogen storage. Finally, the created 3D gelatin scaffold could provide an appropriate matrix for hepatogenic differentiation of hADSCs, which could be considered for liver tissue engineering applications.
•Gelatin cryogel scaffold enhanced attachment and proliferation of hADSCs.•Gelatin cryogel scaffold promoted hepatogenic differentiation of hADSCs.•The 3D network of the Gelatin cryogel scaffold provides a hepatomimetic environment.•A novel and simple strategy for developing the hADSCs based liver tissue engineering.