In vitro coculture

Scope of the method

The Method relates to
  • Animal health
  • Human health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
Caco-2 and HepG2 cells


Method keywords
  • cell culture
  • Cell interactions
  • in vitro digestion
  • Metabolism
Scientific area keywords
  • in vitro cell culture
  • liver metabolism
  • Diabetes research
  • Sugar metabolism
  • Fat metabolism
Method description

This method is used to let cells interact for a better simulation of processes occurring in the body. The rational of the method is that monoculture experiments do not capture the complexity of in vivo interactions between different organs. In the most simple setup (published work Nutrients), Caco-2 cells were seeded on the (in our setup 24-well) transwell insert and HepG2 cells on the (in our setup 5*104 cells in a 24-well) plate below. Caco-2 cells were seeded first. HepG2 cells were seeded in a separate plate upon differentiation of the Caco-2 cells and were put together when HepG2 cells formed a confluent layer. The Caco-2 cells were exposed to sugars and fatty acids, allowing digestion and transport to the HepG2 cells.

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Lab equipment
  • - Coculture plates with inserts
  • - Cell culture flow cabinet
  • - TEER machine or Lucifer Yellow
Method status
  • Published in peer reviewed journal

Pros, cons & Future potential


It increases the relevance of the cell culture model compared to the monoculture variants. Furthermore, it allows to study the effects of nutrients and medicinal compounds on organs that are only in contact with metabolites of these compounds in vivo (the method allows to incorporate digestion, absorption and metabolism).


The upper layer has to be confluent during the entire exposure (has to be checked before and after coculture and after the exposure). Since the cells should also be sufficiently fresh, timing is important. This timing depends on the cell type.


This method could basically be used with any cell type and could even be used with more than 2 types of cells interacting (which requires some adjustment).

Future & Other applications

The method can be applied for all type of cell research, not just in the field of diabetes.

References, associated documents and other information

  • van Laar, A., Grootaert, C., Van Nieuwerburgh, F., Deforce, D., Desmet, T., Beerens, K., & Van Camp, J. (2022). Metabolism and Health Effects of Rare Sugars in a CACO-2/HepG2 Coculture Model. Nutrients, 14(3), 611.
  • Sadeghi Ekbatan, S., Iskandar, M. M., Sleno, L., Sabally, K., Khairallah, J., Prakash, S., & Kubow, S. (2018). Absorption and metabolism of phenolics from digests of polyphenol-rich potato extracts using the Caco-2/HepG2 co-culture system. Foods, 7(1), 8.
  • Lammi, C., Zanoni, C., Ferruzza, S., Ranaldi, G., Sambuy, Y., & Arnoldi, A. (2016). Hypocholesterolaemic activity of lupin peptides: Investigation on the crosstalk between human enterocytes and hepatocytes using a co-culture system including Caco-2 and HepG2 cells. Nutrients, 8(7), 437.
  • Scheers, N. M., Almgren, A. B., & Sandberg, A. S. (2014). Proposing a Caco-2/HepG2 cell model for in vitro iron absorption studies. The Journal of nutritional biochemistry, 25(7), 710-715.

Contact person

Amar van Laar


Ghent University (UGent)
Food Technology, Safety and Health