The human intestines are a critical site, not only for absorption of nutrients and water but also pharmaceutical drugs. In addition to this, it serves as an important barrier protecting the body from pathogens.
3D printing intestinal cells can give the possibility to create more advanced and specific tissue like structures compared to classical cell culture. Printed constructs can be cultured in filter inserts in order to study barrier function, transport of nutrients, bioactive molecules and pharmaceuticals.
The aim of following project was to culture and evaluate the viability of Caco 2 cells bioprinted in CELLINK EPI X ink.
We have printed Caco 2 cells which is a cell line from human epithelial colorectal adenocarcinoma cells.
This cell line has previously been used to study if components of the sigma B regulon, which controls the transcriptional response to stress, in Listeria monocytogenes contribute to cell invasion. It has also been to study the effect of Maslinic acid, a promising chemopreventive agent, on colon cancer cell lines.
Cells can be printed alone, in co-culture or in separate layers or compartments. With the BIO X 3D bioprinter you can print up to three different cell types in one go.
Cells were mixed with EPI X bioink at a concentration of 10 million cells/ml bioink and 30 million cells/ml bioink. Using the BIO X, 3 x 0,6 mm discs were printed in 24-well plates. The printing speed was 10 mm/s, the printing pressure was 3-5 kPa and 0,410 mm/22G conical nozzles were used.
Both cell number and viability were stable between Day 1 and Day 8. Viability was around 80% in all cases. No major differences in viability could be seen between the two seeding densities. The stable numbers indicate that the cells are performing well and that the EPI X bioink could be used for further tests.
EPI X seems to support 3D bioprinted Caco 2 cells. This will provide a basis for further studies using CELLINK EPI X bioink. Caco 2 cells could be printed either alone or together with other cells, for example a mucus secreting cell line in order to generate models of intestinal epithelium.