Alginate-chitosan core-shell microcapsule cultures of hepatic cells in a small scale stirred bioreactor: impact of shear forces and microcapsule core composition

Abstract

A small scale stirred bioreactor was designed and the effect of different agitation rates (30, 60 and 100 rpm) was investigated on HepG2 cells cultured in alginate-chitosan (AC) core-shell microcapsule in terms of the cell proliferation and liver-specific function. The microencapsulated hepatic cells could proliferate well when they were cultured for 10 days at 30 rpm while the cell-laden microcapsules showed no cell proliferation at 100 rpm in the bioreactor system. Albumin production rate, as an important liver function, increased also 1.8- and 1.5- fold under stirring rate of 30 rpm compared to the static culture and 60 rpm of agitation, respectively. Moreover, In comparison with the static culture, about 1.5-fold increment in urea production was observed at 30 rpm. Similarly, the highest expressions of albumin and P450 genes were found at 30 rpm stirring rate, which were 4.9- and 19.2-fold of the static culture. Addition of collagen to the microcapsule core composition (ACol/C) could improve the cell proliferation and functionality at 60 rpm in comparison with the cell-laden microcapsules without collagen. The study demonstrated the hepatic cell-laden ACol/C microcapsule hydrogel cultured in the small scale stirred bioreactor at low mixing rate has a great potential for mass production of the hepatic cells while maintaining liver-specific functions.

Synthesis, optimization, and cell response investigations of natural-based, thermoresponsive, injectable hydrogel: An attitude for 3D hepatocyte encapsulation and cell therapy

“…Hepatocytes cultured in different 3D scaffolds of other studies showed similar enhancements in liver-specific activities (Wu et al, 2014;Wang et al, 2019). The improved microencapsulated cell viability outcomes are consistent with the liver-specific function data, suggesting the benefits of the 3D culture system (Khodabakhshaghdam et al, 2021). This indicates that the 3D culture system with a specific hydrogel combination had no cytotoxic effect on cell metabolic activity.…”

Current strategies with implementation of three-dimensional cell culture: the challenge of quantification

“…The cells in these systems are typically under sheer stress, allowing a high mass transfer rate to be achieved throughout the cultures [37][38][39]. Bioreactors have been successfully implemented for a variety of cell types, including heart, muscle, liver, embryonic stem cells and mesenchymal stem cells [40][41][42][43][44][45]; while showing great potential in the toxicity testing of potential therapeutic drugs [46][47][48].…”

Design considerations of benchtop fluid flow bioreactors for bio-engineered tissue equivalents in vitro

“…The stir speed and shape of the stirrer allows for significant control over the mass transfer and shear stress in the system. Variation of the stir speed in this manner has been shown to impact the levels of different ECM components secreted by chondrocytes [57] and affect properties such as albumin secretion in HepG2 hepatocellular carcinoma cells [58] . One of the most common uses in tissue engineering is for the expansion of stem cells by maintaining stem cell pluripotency and offering an alternative to methods such as feeder layers [59] , [60] , [61] .…”