Evaluation of a galactose‐carrying gelatin sponge for hepatocytes culture and transplantation

Abstract

This study proposes a new three-dimensional culture of mouse hepatocytes in a porous galactose-carrying modified gelatin sponge matrix. The modification of gelatin using galactose residues significantly increased the attachment of hepatocytes on the substrate. A modified gelatin sponge with lactobionic acid (MGLA) was prepared to increase the specific interaction between the hepatocytes and the matrix. Hepatocytes cultured in a three-dimensional MGLA sponge released much less lactate dehydrogenase than those cultured on a collagen Type I-coated monolayer. Moreover, the survival rate of hepatocytes cultured on an MGLA sponge was longer than the survival rate of hepatocytes cultured on a collagen Type I-coated monolayer. Hepatic specific metabolic functions, namely, the secretion of serum albumin and the synthesis of urea, were well maintained and promoted by spheroidal hepatocytes formed in the MGLA sponge.

Optimization of 3-D hepatocyte culture by controlling the physical and chemical properties of the extra-cellular matrices

“…This is in contrast to Mooney et al's [29] study on 2-D ECM-coated dishes, where they found that increasing collagen coating density down-regulated hepatocyte functions with corresponding cell spreading. While galactose incorporation onto biomaterials has been shown to stimulate hepatocyte functional maintenance [15][16][17]30], our results clearly show that chemical support itself is inadequate. We propose a model in which both physical and chemical supports of the ECM are important for optimal hepatocyte functions in vitro.…”

“…Physical control of the collagen nanofibres was achieved by finetuning the complex-coacervation reaction between methylated collagen and terpolymer of hydroxylethyl methacrylate-methyl methacrylate-methylacrylic acid (HEMA-MMA-MAA) and quantitative characterization using a back-scattering confocal microscopy assay. We further enhanced the chemical properties of the collagen nano-fibres by incorporating galactose onto collagen, which specifically interacts with the asialoglycoprotein receptor (ASGPR) on hepatocytes to promote their functional maintenance [15][16][17]. By correlating fibres with a range of different physicochemical parameters to hepatocyte functions, we have identified a specific combination of methylated and galactosylated collagen nano-fibres with optimal physicochemical characteristics for maintaining hepatocyte functions in vitro.…”

“…To further promote hepatocyte functions, we galactosylated collagen to provide specific chemical interactions between galactose ligands conjugated to collagen and the ASGPR on hepatocytes [15][16][17]. Carboxyl groups on collagen were reacted with 1-N-(lactobionic acyl)-ethylenediamine to incorporate galactose onto the collagen chains (Fig.…”

Myogel, a Novel, Basement Membrane-Rich, Extracellular Matrix Derived from Skeletal Muscle, Is Highly Adipogenic in vivo and in vitro

“…The opportunities for these diverse applications have led to the creation of hydrogels from synthetic and/or naturally occurring polymers. Examples include collagen or gelatin sponges [Hong et al, 2002;Kimura et al, 2003], poly(ethylene glycol)diacrylate with added peptide sequences [Mann and West, 2002;Gobin and West, 2003;Lutolf et al, 2003], hyaluronan-gelatin hydrogels [Lui et al, 2004], polyethylene glycol/polylactic co-glycolic acid [Jeong et al, 2002;Drury and Mooney, 2003] and polyvinyl alcohol [Veronese et al, 2001;Schmedlen et al, 2002]. Other substances formulated as hydrogels include chitosan from crustacean shells, fibrin (a blood-clotting polymer), alginate extracted from seaweed [Eiselt et al, 2000;Kuo and Ma, 2001;Chen et al, 2003;Wang et al, 2003] and Puramatrix, a synthetic polypeptide hydrogel [Bokhari et al, 2005].…”

Polymers in Tissue Engineering

“…The first examples of functional transplantable liver-like structures used prevascularized, non-degradable polyvinyl alcohol sponges to accommodate transplanted hep-atocytes with limited success [131,132]. Modified gelatin sponges with lactobionic acid (MGLA) have also been used to support mouse hepatocyte growth [133]. Microhydrogels made of fibrinogen attached to poly(ethylene glycol) (PEG)-diacrylate side chains were used as a cell carrier for intravascular transplantation of hepatocytes in a rat model [134].…”