Mesenchymal Stem Cells Differentiation Hepatocyte-like Cells 3D Microenvironment

Objectives: Our study aimed to select the optimal growth factor cocktail for the differentiation of MSCs into hepatocyte-like cells (HLCs) in the absence and presence of a 3D microenvironment. Methods: Ninety ml BM was aspirated from the iliac bone for separation of MSCs. This study was conducted on 20 patients with advanced liver cirrhosis, divided into two groups. Group I; MSCs were plated onto 96-well plates without microenvironment (control group). Group II; MSCs were plated onto 96-well plates with a 3D microenvironment. Surface expression of CD271, CD29, and CD34 were analyzed using flow cytometry. MSCs were differentiated in-vitro into HLCs by adding four growth factor cocktails in presence and absence of 3D microenvironment. Hepatogenesis was assessed by immunohistochemical analysis of OV6, AFP, albumin, and cytokeratin 18 expressions. Results: There was a statistically significant increase in the expression of CD271 and CD29 after MSCs culture (P <0.001). A heterogeneous cell population composed of MSCs and differentiated HLCs was encountered (40% hepatocytes and 60% MSCs). Furthermore, our results showed that growth factor cocktails one and two gave the best results for differentiation of MSCs into HLCs. Conclusion: MSCs could be a feasible, readily available, and novel source for differentiation into HLCs for future therapeutic implications.

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