Decellularized liver matrix-modified chitosan fibrous scaffold as a substrate for C3A hepatocyte culture.
A bioreactor filled with functional hepatocytes is an important part of the bio-artificial liver device. However, it is a difficult task to maintain a sufficient number of cells and hepatocellular function is active. In this work, we developed a promising scaffold for hepatocyte culture by coating the pork liver extracellular matrix (ECM) in chitosan (CTS) fabric.
Porcine hearts were decellularized using 1% Triton X-100. ECM dissolved CTS heart was immobilized on the fiber surface through cross linking of ECM and CTS with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-Hydroxysuccinimide (NHS). Then scaffold characterized by Fourier transformed infrared spectroscopy attenuated total reflection mode (ATR-FTIR), X-photoelectron spectroscopy (XPS) and water contact angle measurements.
Efficacy scaffold modified to maintain hepatocyte C3A adhesion, proliferation, and functionality in vitro bioactivity detected. FTIR spectra and XPS show their ECM CTS coating on the fabric surface. Covalent coating significantly increases the efficiency of binding between ECM and CTS fabric, compared with a coating by physical absorption.
Furthermore, the C3A hepatocytes cultured on the scaffold showed an increase bioactivity coated cells and liver-specific functions, such as albumin secretion and synthesis of urea, compared to those cultivated untreated scaffold (p <0.05). As a carrier of culture hepatocytes promising, ECM coated fabric CTS can be applied in biological artificial liver reactors.
a comparative study between the lactose-silk fibroin and extracellular matrix conjugate as a substrate for the culture of human induced pluripotent stem cell-derived hepatocytes.
Human induced pluripotent stem cells (hiPSC) -derived hepatocytes a cell source attractive alternative to primary human hepatocytes for studies network regeneration.This gift an app from lactose-silk conjugate (Lac-CY-SF) bearing β-galactose residue as substrate hiPSC derived hepatocytes for culture.
Comparison hiPSC derived hepatocytes were cultured on three different substrates; Lac-CY-SF conjugates, Matrigel and type I collagen is performed.Cell morphology, viability, maturation and function of albumin secretion was assessed by phase contrast microscopy, tetrazolium based colorimetric assay, immunofluorescence staining and characteristics of the enzyme-linked immunosorbent assay.
Morphological of cell- cells were cultured on a conjugate resemble those in Matrigel throughout the 6-day culture period. The number of viable cells cultivated in the conjugate is comparable to that in Matrigel at day 2 and 6. The protein expression of hepatocyte markers of adulthood, asialoglycoprotein receptor 1 and albumin, the cells are cultured on a conjugate resembles that in cells cultured in collagen on days 2 and 6.Connection error.
Albumin function of secretion per cell cultured on a conjugate higher than in collagen and comparable to that in the limited Matrigel.These results showed that Lac-CY-SF conjugates may be useful as Matrigel and collagen for cultivation hiPSC derived hepatocytes.