Benefits of the attachment
- Transwells in which cells and microbe are cultured can be separated from each other
- The damage caused by microbe to cells is greatly reduced by the prevention of contact between them
- Only transwells for microbial culture can be made anaerobic conditions using liquid paraffin
- Multiple specimens can be cultured simultaneously using existing plates and transwells
Background and Technology
The study of cell-microbe interactions is particularly important for understanding the gut environment. Cell-microbe in vitro co-culture systems as a research tool have many advantages, such as the ability to control culture conditions and directly measure cell growth and metabolic changes. Several in vitro co-culture systems have been proposed, but the focus has been solely on assessing the effects on the cultured cells. This has led to the following limitations: (1) monitoring of medium sampling and microbial growth is difficult; (2) the systems tend to be large and complex and lack simplicity and uniformity; (3) it is difficult to culture cells and micro-organisms under anaerobic conditions when they are cultured in the same well; and (4) The direct action of micro-organisms causes significant damage to cells and weakens intercellular adhesion.
The inventors invented an attachment for placing transwells for cell culture in 6-/12-well plates and overlaying transwells for microbiological culture with a smaller diameter (see Figure B below). A simple co-culture system using commercially available plates and wells allows (1) free sampling of the medium during co-culture, (2) monitoring of microbial and cultured cell growth over time, (3) aerobic-anaerobic co-culture, (4) simple operation and simultaneous assessment of multiple samples. The following features are available.
Data
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- In an in vitro co-culture system with attachments (—-: joint plate), anaerobic Bifidobacterium (B. bifidum) were cultured in small transwells (12 well insert), and the surface of the culture medium was coated with liquid paraffin, while large transwells (6 well insert) were cultured with MDCK cells (A). (C) is an actual photograph.
- The barrier function and viability of MDCK cells were comparable to those of normal MDCK cultures, and the growth of bifidobacteria under sealed conditions was approximately 1.5 times higher than under unsealed conditions.
Patent & Publication
Design pending (unpublished)
Publishment：https://doi.org/10.1016/j.jbiosc.2023.03.008
Researcher
Dr. Yoshihiro Umehara, Dr. Hidenori Aoyagi (University of Tsukuba)
Project.WL-04969