Cells can be cultured in the lab for experimental purposes and some types of cells are cultured in suspension in a culture medium, while others are adherent to surfaces in cell culture flasks. After the cells have divided and require more space for further growing, they are detached from the cell culture surface and placed in new culture flasks in fewer numbers. This is called a passage of the cells, and different types of cells tolerate different numbers of passages in culture. The project will investigate how human keratinocytes behave through cell culture passaging, by comparing passage 0 cells with cells in passage 1 and passage 5. The RNA-expression of the cells will be investigated with the aid of q-PCR and a panel of 90 genes will be examined. Changes in gene expression can indicate how the cells adapt to in vitro culture, and the project seeks to add to the knowledge on how good in vitro methodology is, for obtaining results that are transferable to in vivo circumstances for keratinocytes.
We wish to investigate what the things we do in the lab actually say about reality, how keratinocyte gene expression changes during passaging in culture, and how much we need to take those changes into consideration in experimental design