Three-dimensional high-density culture of HepG2 cells in a 5-ml radial-flow bioreactor for construction of artificial liver

Liver endothelial cells are important components of the tissue along the hepatic sinusoid. They are responsible for microcirculation in the liver and scavenger functions. It would therefore be important to include these cells in any hybrid type of artificial liver in addition to hepatocytes. However, it is difficult to culture these cells in vitro. The development of a liver endothelial cell line, which maintains the characteristics of the primary culture, would thus be of great benefit in the development of an artificial liver. In the present study we established immortalized liver endothelial cells from the liver of an H-2Kb-tsA58 transgenic mouse, which harbors the SV40 TAg gene. Hepatic sinusoidal cells isolated from H-2Kd-tsA58 mouse proliferated In the presence of γ-interferon at 33°C. Four clones were established, out of which clone M1 had the highest amounts of PGI2 production, as well as plasminogen activator activity and internalized acetylated low density lipoprotein. On culture dishes the M1 cells grew individually and spread. Sieve plates on the cell surface were not readily visible, but small pores were detected under electron microscopic observation. These results suggest that M1 clone cells originated from liver endothelial cells. Moreover it was possible to culture the immortalized liver endothelial cells in a radial-flow bioreactor for 5 days, with a maximum 6-keto prostaglandin F1α production of 25 μg per day. This suggests that immortalized liver endothelial cells and a radial-flow bioreactor can prove useful tools in the development an artificial liver.

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Production and release of infectious hepatitis C virus from human liver cell cultures in the three-dimensional radial-flow bioreactor

Virology ◽

10.1016/s0042-6822(03)00383-0 ◽

2003 ◽

Vol 314 (1) ◽

pp. 16-25 ◽

Cited By ~ 40

Author(s):

Hideki Aizaki ◽

Seishi Nagamori ◽

Mami Matsuda ◽

Hayato Kawakami ◽

Osamu Hashimoto ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Cell Cultures ◽

Liver Cell ◽

Human Liver ◽

Radial Flow ◽

Three Dimensional ◽

Infectious Hepatitis ◽

Human Liver Cell ◽

Radial Flow Bioreactor

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Three-dimensional multiscale fiber matrices: development and characterization for increased HepG2 functional maintenance for bio-artificial liver application

Biomaterials Science ◽

10.1039/c7bm00963a ◽

2018 ◽

Vol 6 (2) ◽

pp. 280-291 ◽

Cited By ~ 13

Author(s):

Surendra Kumar Verma ◽

Akshay Modi ◽

Jayesh Bellare

Keyword(s):

Hepg2 Cells ◽

Three Dimensional ◽

Characteristic Functions ◽

Artificial Liver ◽

Fiber Matrices ◽

One Step

One-step development of three-dimensional multiscale fiber matrices to enhance attachment, proliferation, and characteristic functions of HepG2 cells.

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Reconstruction of Liver Organoid Using an Apatite-Fiber Scaffold, a Radial-Flow Bioreactor and FCL-4 Cells of Hepatocyte Model

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.361-363.1165 ◽

2007 ◽

Vol 361-363 ◽

pp. 1165-1168 ◽

Cited By ~ 1

Author(s):

Mamoru Aizawa ◽

A. Hiramoto ◽

H. Maehashi ◽

Tomokazu Matsuura

Keyword(s):

Tissue Engineering ◽

Single Crystal ◽

Bone Tissue Engineering ◽

Radial Flow ◽

Three Dimensional ◽

3D Culture ◽

High Porosity ◽

Fiber Scaffolds ◽

Radial Flow Bioreactor ◽

Culture Of Hepatocytes

We have previously developed apatite-fiber scaffolds (AFSs) for bone tissue engineering using single-crystal apatite fibers and carbon beads. In the present investigation, we examined the possibility of reconstruction of a liver organoid via three-dimensional (3D) culture of hepatocytes using the AFSs and the radial-flow bioreactor (RFB), aiming to apply the scaffold as a matrix for regeneration of a real organ. FLC-4 cells were used as a model of hepatocyte. The cells were well-viable in the RFB settled with AFSs over a period for 28 d, compared with the cases of cellulose beads and apatite beads with high porosity of 85%. We conclude that the present AFS may be a promising scaffold for tissue engineering of liver.

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Influence of Culture Period on Osteoblast Differentiation of Tissue-Engineered Bone Constructed by Apatite-Fiber Scaffolds Using Radial-Flow Bioreactor

International Journal of Molecular Sciences ◽

10.3390/ijms222313080 ◽

2021 ◽

Vol 22 (23) ◽

pp. 13080

Author(s):

Kitaru Suzuki ◽

Jun Fukasawa ◽

Maiko Miura ◽

Poon Nian Lim ◽

Michiyo Honda ◽

...

Keyword(s):

Osteoblast Differentiation ◽

Radial Flow ◽

Bone Marrow Cells ◽

Three Dimensional ◽

3D Cell Culture ◽

Bone Diseases ◽

Culture Period ◽

Alizarin Red ◽

Fiber Scaffolds ◽

Radial Flow Bioreactor

With the limitation of autografts, the development of alternative treatments for bone diseases to alleviate autograft-related complications is highly demanded. In this study, a tissue-engineered bone was formed by culturing rat bone marrow cells (RBMCs) onto porous apatite-fiber scaffolds (AFSs) with three-dimensional (3D) interconnected pores using a radial-flow bioreactor (RFB). Using the optimized flow rate, the effect of different culturing periods on the development of tissue-engineered bone was investigated. The 3D cell culture using RFB was performed for 0, 1 or 2 weeks in a standard medium followed by 0, 1 or 2 weeks in a differentiation medium. Osteoblast differentiation in the tissue-engineered bone was examined by alkaline phosphatase (ALP) and osteocalcin (OC) assays. Furthermore, the tissue-engineered bone was histologically examined by hematoxylin and eosin and alizarin red S stains. We found that the ALP activity and OC content of calcified cells tended to increase with the culture period, and the differentiation of tissue-engineered bone could be controlled by varying the culture period. In addition, the employment of RFB and AFSs provided a favorable 3D environment for cell growth and differentiation. Overall, these results provide valuable insights into the design of tissue-engineered bone for clinical applications.

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