Metabolic capability of upcyte human hepatocytes in long-term sandwich culture and utility for clearance prediction

Hepatocytes in long-term cultures represent a promising approach to preserve liver function under standard culture conditions. Hepatocyte cultures as the key components in an extracorporeal artificial liver (EAL) in the treatment of hepatic insufficiency, would be a great advantage. However, one of the numerous unsolved problems is the limitation of the surface area of a future EAL. To decrease the dimensions of same, we modified the cell immobilization technique by placing a second layer of immobilized human hepatocytes onto a layer of pre-immobilized hepatocytes creating a “sandwich immobilization” (SI) system. Immobilization and sandwich immobilization were compared over an investigation period of 30 days: functional performance mirrored by cholinesterase (CHE) and albumin secretion showed remarkable differences only in the course of the first week, whereas we found almost no differences from day 8 on. The total DNA-values on days 0, 1, 7, 14, 21 and 30 varied strongly after the first week but were very similar up to day 30. Finally, it appears disadvantageous to enlarge number/cm2 of (human) hepatocytes in long-term cultures or for application in an EAL by means of sandwich immobilization.

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Long-Term Culture of Rat Hippocampal Neurons at Low Density in Serum-Free Medium: Combination of the Sandwich Culture Technique with the Three-Dimensional Nanofibrous Hydrogel PuraMatrix

PLoS ONE ◽

10.1371/journal.pone.0102703 ◽

2014 ◽

Vol 9 (7) ◽

pp. e102703 ◽

Cited By ~ 20

Author(s):

Ai Kaneko ◽

Yoshiyuki Sankai

Keyword(s):

Hippocampal Neurons ◽

Three Dimensional ◽

Culture Technique ◽

Low Density ◽

Serum Free Medium ◽

Free Medium ◽

Serum Free ◽

Long Term Culture ◽

Sandwich Culture

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A Long-Term Culture of Human Hepatocytes Which Show a High Growth Potential and Express Their Differentiated Phenotypes

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.1999.0288 ◽

1999 ◽

Vol 256 (1) ◽

pp. 184-191 ◽

Cited By ~ 57

Author(s):

Hiroshi Hino ◽

Chise Tateno ◽

Hajime Sato ◽

Chihiro Yamasaki ◽

Shigeru Katayama ◽

...

Keyword(s):

High Growth ◽

Human Hepatocytes ◽

Growth Potential ◽

Long Term Culture ◽

High Growth Potential

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Long‐term maintenance of functional primary human hepatocytes using small molecules

FEBS Letters ◽

10.1002/1873-3468.13582 ◽

2019 ◽

Vol 594 (1) ◽

pp. 114-125 ◽

Cited By ~ 2

Author(s):

Takeshi Katsuda ◽

Masaki Kawamata ◽

Ayako Inoue ◽

Tomoko Yamaguchi ◽

Maki Abe ◽

...

Keyword(s):

Small Molecules ◽

Human Hepatocytes ◽

Primary Human Hepatocytes ◽

Term Maintenance

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Long-term functional maintenance of primary human hepatocytes in vitro

Science ◽

10.1126/science.aau7307 ◽

2019 ◽

Vol 364 (6438) ◽

pp. 399-402 ◽

Cited By ~ 36

Author(s):

Chengang Xiang ◽

Yuanyuan Du ◽

Gaofan Meng ◽

Liew Soon Yi ◽

Shicheng Sun ◽

...

Keyword(s):

Cell Biology ◽

Expression Profiles ◽

Antiviral Drug ◽

Gene Expression Profiles ◽

Global Gene Expression ◽

Human Hepatocytes ◽

Hbv Infection ◽

Primary Human Hepatocytes

The maintenance of terminally differentiated cells, especially hepatocytes, in vitro has proven challenging. Here we demonstrated the long-term in vitro maintenance of primary human hepatocytes (PHHs) by modulating cell signaling pathways with a combination of five chemicals (5C). 5C-cultured PHHs showed global gene expression profiles and hepatocyte-specific functions resembling those of freshly isolated counterparts. Furthermore, these cells efficiently recapitulated the entire course of hepatitis B virus (HBV) infection over 4 weeks with the production of infectious viral particles and formation of HBV covalently closed circular DNA. Our study demonstrates that, with a chemical approach, functional maintenance of PHHs supports long-term HBV infection in vitro, providing an efficient platform for investigating HBV cell biology and antiviral drug screening.

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