Alginate‐encapsulated HepG2 Cells in a Fluidized Bed Bioreactor Maintain Function in Human Liver Failure Plasma

Abstract

Alginate-encapsulated HepG2 cells cultured in microgravity have the potential to serve as the cellular component of a bioartificial liver. This study investigates their performance in normal and liver failure (LF) human plasma over 6-8 h in a fluidized bed bioreactor. After 8 days of microgravity culture, beads containing 1.5 x 10(9) cells were perfused for up to 8 h at 48 mL/min with 300 mL of plasma. After exposure to 90% LF plasma, vital dye staining showed maintained cell viability, while a 7% increase in lactate dehydrogenase activity indicated minimal cell damage. Glucose consumption, lactate production, and a 4.3-fold linear increase in alpha-fetoprotein levels were observed. Detoxificatory function was demonstrated by quantification of bilirubin conjugation, urea synthesis, and Cyp450 1A activity. These data show that in LF plasma, alginate-encapsulated HepG2 cells can maintain viability, and metabolic, synthetic, and detoxificatory activities, indicating that the system can be scaled-up to form the biological component of a bioartificial liver.

Storage Temperatures for Cold-Chain Delivery in Cell Therapy: A Study of Alginate-Encapsulated Liver Cell Spheroids Stored at −80°C or −170°C for Up to 1 Year

“…Our group has focused on development of encapsulated liver cell spheroids (ELS) as a part of a bioartificial liver (BAL) to bridge the gap either to transplant or to spontaneous recovery in patients suffering from acute liver failure (ALF). 1 The system requires conditioning culture for up to 10 days to allow formation of 3D ELS that are sufficiently functional to treat ALF. ALF, however, can develop within 1-2 days and requires immediate support.…”

Encapsulated human hepatocellular carcinoma cells by alginate gel beads as an in vitro metastasis model

“…Most importantly, after spheroids formed within the gels, they can be easily retrieved for further testing by simply adding citrate sodium solution without cell damage in several minutes [15]. ALGs have been widely applied to tissue engineering [16] and tumor research [17,18]. Previous studies show that encapsulation of liver cancer cells into ALG scaffolds may actually enhance cell survival, proliferation [19,20], secretion of ECM proteins [21] and tumor malignancies [22].…”

3 dimensional cell cultures: a comparison between manually and automatically produced alginate beads

“…In this respect, three dimensional cell cultures are an important strategy for compound screenings (Rimann and Graf-Hausner 2012), tissue engineering (Coward et al 2009) and investigation of tumors (Rokstad et al 2012). Three dimensional cell cultures enable the mimic of in vivo conditions.…”

“…The low density alginate beads (0.5 x 10 6 cells/ml alginate) showed an increased proliferation until day ten. The higher proliferation rates of cells in three dimensional cell culture systems is probably be caused by lower shear stress (Coward et al 2009). However, the proliferation of high density alginate bead cultures (2 9 10 6 cells/ ml alginate) was decreased till day 14 (Moshaverinia et al 2013b).…”

“…Some research groups investigated the automated cell encapsulation for different applications. The investigation of the proliferation potential of HepG2 cells embedded in alginate showed an increased proliferation and a-1 antitrypsin concentration in a rotary cell culture system over the time (Coward et al 2009). Kim et al (2013) evaluated the encapsulation of human embryonic stem cells on the differentiation potential to dopamine neurons.…”