This semester Prof. Isaacson and I are working with HEK 293 cells instead of 293t cells. In the past, we worked with 293 t cell line and study the localization of the TORC family and tax protein in these. HEK 293 is a cell line derived from human embryonic kidney cells in culture and they differ from the 293t cells because they do not contain the SV40 large T antigen that is capable of inducing malignant transformation. The purpose of working with the HEK 293 this semester is to study localization of the TORC family in the cells and afterwards compare it to the localization of these proteins in the 293 t cells. We are interested in seeing if malignant transformation of the cells affects the localization of the proteins (TORC).
We are currently doing immunofluorescence to study localization but so far this semester they have all been a fail (3 so far). The HEK 293 cells do not grow as fast as the 293 t cells and do not adhere as well either. This has proven to be a problem as they are harder to maintain after every split and they do not adhere to the glass cover slips very well. In fact, I had to dispose of two plates because they did not adhere at all to the glass cover slips. If they do not adhere, I am not able to perform any experiment on them and this has been my biggest challenge this semester. I am hoping that by treating the glass cover slips with a special chemical the cells will be able to adhere and I will be able to perform my experiments.