Author/Authors :
Talaei-Khozani Tahereh نويسنده , Azarnia Mahnaz نويسنده Department of Biology, School of Science, Tarbiat Moalem University, Tehran, Iran , Jalili Firoozinezhad Sasan نويسنده Department of Regenerative Biomedicine at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran , Jaberipour Mansoureh نويسنده Institute Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran. , Bonakdar Shahin نويسنده National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, IR Iran , Rajabi-Zeleti Sareh نويسنده , Aleahmad Fatemeh نويسنده Department of Animal Biology, Faculty of Biological
Sciences, Kharazmi University, Tehran, IR Iran , Sani Mahsa نويسنده Tissue Engineering Lab, Shiraz Medical School, Shiraz
University of Medical Sciences, Shiraz, IR Iran , Heshmat-Azad Sanaz نويسنده Department of Chemistry, Isfahan University of Technology,
Isfahan, IR Iran
Abstract :
Background Recapitulating the native cell niche and extracellular
matrix (ECM) architecture in vitro helps reconstruct injured tissues.
Collagen I and heparin are two important constituents of the ECMs, which
play crucial roles in regulating cell behaviors. Specifically in the
liver, these components can affect the differentiation and functionality
of the cells. Objectives The aims of this study were to first fabricate
and characterize a heparin/collagen scaffold and then investigate the
scaffold’s efficiency in directing the differentiation of Wharton’s
jelly-derived mesenchymal stem cells (WJ-MSCs) towards hepatocyte.
Methods After fabricating the rat tail collagen I sponge-shaped
scaffolds, heparin was chemically immobilized on the scaffolds using
N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC) and
N-hydroxysuccinimide (NHS). The scaffold chemical characteristics and
architecture were evaluated by Fourier-transformed infrared (FTIR)
spectroscopy and scanning electron microscopy (SEM), respectively. In
the next step, Wharton’s jelly-derived mesenchymal stem cells were
seeded on the scaffolds and cell viability and morphology were assessed
using MTT assay and SEM, respectively. Moreover, followed by exposing
the WJ-MSCs to the hepatogenic media for 3 weeks, liver-specific marker
expression and indocyanine green (ICG) clearance tests were performed.
Results The data showed that 0.25 mg/mL heparin had no detrimental
effects on the cell viability and proliferation compared to the observed
effects in non-heparinized conditions. SEM micrographs showed that while
immobilizing heparin had no considerable effect on the porosity of the
scaffolds, the mean value of the pore sizes of heparinized sponges was
higher than that of non-heparinized ones. The hepatocyte differentiation
appeared to be enhanced in the cells cultured in heparinized sponges as
indicated by higher percentage of the cells expressing cytokeratin 19
and albumin as well as improved indocyanine green clearance levels.
Conclusions Heparin/collagen scaffold serves as a good platform for
promoting differentiation into hepatocytes and therefore, it has the
potential to be considered for drug discovery and liver regenerative
medicine.
