title: Improvement of mesenchymal stem cell differentiation into the endoderm lineage by four step sequential method in biocompatible biomaterial.
abstract: BACKGROUND
  The goal of the study described here, was to investigate the potential of umbilical cord derived mesenchymal stem cell (UC-MSCs) into hepatocyte like cells in a sequential 2D and 3D differentiation protocols as alternative therapy.
  
  
  METHODS
  Mesenchymal stem cells (MSCs) were isolated from the umbilical cord (UC) and CD markers were analyzed by flow cytometry. For hepatic differentiation of UC-MSCs, cells were induced with a sequential 4-step protocol in 3D and 2D culture system. Urea concentration and albumin secretion into the culture medium was quantified by ELISA. Gene expression levels of AFP, ALB, and CK18 were determined by RT-PCR. Data were statistically analyzed by the SPSS software. The difference between the mean was considered significant when p < 0.05.
  
  
  RESULTS
  Growth factor dependent morphological changes from elongated fibroblast-like cells to round epithelial cell morphology were observed in 2D culture. Cell proliferation analysis showed round-shaped morphology with clear cytoplasm and nucleus on the alginate scaffold in 3D culture. The mean valuses of albumin production and urea secretion were significantly higher in the 3D Culture system when compared with the 2D culture (p = 0.005 vs p = 0.001), respectively. Treatment of cells with TSA in the final step of differentiation induced an increased expression of CK18 and a decreased expression of αFP in both the 3D and 2D cultures (p = 0.026), but led to a decreased albumin gene expression, and an increased expression in the 2D culture (p = 0.001).
  
  
  CONCLUSIONS
  Findings of the present study indicated that sequential exposure of UC-MSCs with growth factors in 3D culture improves hepatic differentiation.
authors.affiliation: Cellular and Molecular Research Center (CMRC), Department of Anatomical Science, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences (AJUMS), Ahvaz, Iran.
authors.forename: Saeed
authors.initials: S
authors.lastname: Azandeh
authors.affiliation: School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran.
authors.forename: Anneh
authors.initials: A
authors.lastname: Mohammad Gharravi
authors.affiliation: Cellular and Molecular Research Center (CMRC), Department of Anatomical Science, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences (AJUMS), Ahvaz, Iran.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Orazizadeh
authors.affiliation: Cancer, Petroleum and Environmental Pollutants Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Khodadi
authors.affiliation: Cellular and Molecular Research Center (CMRC), Department of Anatomical Science, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences (AJUMS), Ahvaz, Iran.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Hashemi Tabar
keywords: 3D culture; Liver; Mesenchymal stem cell; Umbilical cord