Home › Microbiology › "Liver-on-a-Chip" Cultures of Primary Hepatocytes and Kupffer Cells for Hepatitis B Virus Infection
Microbiology JoVE (Open Access) Citable · DOI

"Liver-on-a-Chip" Cultures of Primary Hepatocytes and Kupffer Cells for Hepatitis B Virus Infection

DOI: 10.3791/58333-v
What you'll learn
  • ✓ Assemble and equilibrate liver-on-a-chip microfluidic plates
  • ✓ Thaw and seed primary hepatocytes into monoculture systems
  • ✓ Establish long-term co-cultures of hepatocytes and Kupffer cells
  • ✓ Perform HBV infection experiments in 3-D liver tissue models
Protocol

The goal of this protocol is to provide a step-by-step guide to perform 3-D "liver-on-a-chip" infection experiments with the hepatitis B virus.

Difficulty
advanced
Total time
~5–7 days (including cell thawing, seeding, equilibration, and infection timeline)
Model organism
Primary human hepatocytes, Kupffer cells
Biosafety
BSL-2

Steps

1
Assemble and equilibrate microfluidic liver-on-chip plates

Prepare and assemble the organ-on-chip device components, then equilibrate the system with appropriate media to establish stable microfluidic conditions prior to cell seeding.

▶ 01:38
2
Thaw and seed primary hepatocytes into monocultures

Thaw cryopreserved primary hepatocytes and seed them into the microfluidic chamber at defined density to establish initial hepatocyte monoculture.

▶ 04:36
3
Culture hepatocytes and Kupffer cells in co-culture systems

Maintain long-term co-cultures of primary hepatocytes and Kupffer cells in the liver-on-a-chip device under controlled microfluidic conditions to generate physiologically relevant tissue architecture.

▶ 07:38
4
Infect hepatocyte cultures with hepatitis B virus

Introduce hepatitis B virus to the established hepatocyte and Kupffer cell co-cultures and monitor infection outcomes within the 3-D organ-on-chip system.

▶ 07:38
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