Cellular behaviour of hepatocyte-like cells from nude mouse bone marrow-derived mesenchymal stem cells on galactosylated poly(D,L-lactic-co-glycolic acid)

Abstract

Previously, the galactosylation of poly(d,l-lactic-co-glycolic acid) (PLGA) surface was accomplished by grafting allylamine (AA), using inductively coupled plasma-assisted chemical vapour deposition (ICP-CVD) and conjugating lactobionic acid (LA) with AA via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide (EDC/NHS) activation for hepatic tissue-engineering purposes. As a continuation study, the cellular behaviour of hepatocyte-like cells (HLCs) on the surface of the galactosylated PLGA were investigated. Nude mouse bone marrow-derived mesenchymal stem cells (MSCs) were cultured under hepatogenic conditions and the differentiated cells were characterized by reverse-transcription polymerase chain reaction (RT-PCR), immunofluorescence and periodic acid-Schiff (PAS) staining. Galactosylated PLGA enhanced the proliferation rate of HLCs compared to the control; HLCs on the surface of the sample became aggregated and formed spheroids after 3 days of culture. A large number of cells on the surface of the sample exhibited increased liver-specific functional activities, such as albumin and urea secretions. In addition, multicellular spheroids in the sample strongly expressed phospholyated focal adhesion kinase (pFAK) (cell-matrix interactions), E-cadherin (cell-cell interactions) and connexin 32 (Cox32; gap junction).

Visible Light-Cured Glycol Chitosan Hydrogel Containing a Beta-Cyclodextrin-Curcumin Inclusion Complex Improves Wound Healing In Vivo

“…H&E and Masson’s trichrome staining are histopathological techniques for analyzing cellular behavior and collagen synthesis during wound healing, respectively [ 38 , 39 ]. H&E staining is the most commonly used technique in histopathological studies; however, it cannot be used for distinguishing important histopathological changes during wound healing, such as collagen synthesis [ 38 , 39 ]. Masson’s trichrome staining was adopted to analyze collagen synthesized during wound healing [ 38 ].…”

The multiple functional roles of mesenchymal stem cells in participating in treating liver diseases

“…Furthermore, on histological evaluation, they found that number of proliferating hepatocytes and sinusoidal endothelial cells in the MSC‐CM treatment group had increased by 1.2‐ and 1.6‐fold, respectively . It is believed that MSC secretions contain a number of trophic molecules, including soluble ECM glycoproteins, cytokines and growth factors . It remains unknown what specific mediators present in MSC‐CM are responsible for the reduction in cell death and stimulation of liver regeneration.…”

Connexin‐Based Channels in the Liver

“…This zonation pattern increases and upon complete polarization of the hepatocytes, Cx26 is hereby primarily present at the periportal area due to presence of glucagon, which induces Cx26 mRNA expression (18,60). In accordance with the differentiation expression patterns of Cx proteins seen in stem cell-based regenerative processes and during gestation, Cx32 is frequently used as a functional hepatic marker in stem cell-based hepatocyte differentiation studies (38,70,97,140,160,205). The aim of these studies is to form functional hepatocytes from stem cells, such as umbilical cord matrix stem cells, peritoneal adipose mesenchymal stem cells, human embryonic stem cells or oval cells, and to use differentiated cells as in vitro models or for therapeutic purposes (21,134,153,173).…”