Over the course of the past week, we ran multiple tests on the viability of the cells using NanoEntek’s cell counter. We were hoping for the 20% viability of the cells, previously measured, to rise to an ideal 70%. One hypothesis was that the cell counter was not working properly, however, after upon review in the inverted microscope (manual cell counting), we determined that the cells were far too small to be seen even under 40x magnification.
The cells however continually multiplied, whether it was our initial NBT-ii cell line or another organism that multiplied we do not know. This was apparent through observation of the cloudiness of the complete growth medium inside of the T-75 flasks.
(Image of what the NBT-ii cells should look like)
The cloudiness of the media after multiple days typically shows that the cells are ready for splitting/subculturing meaning that they are dividng and splitting. Because of these factors I contacted, NanoEntek’s United States office and asked if the cell counting procedures I was carrying out were correct. They proceeded to reach out to their Hong Kong offices, to answer my questions. It soon became clear that I was carrying out the proper procedures in the cell counter and the viability of my cells was very low. As a result of the lack of viability of my cells I reached out to ATCC’s technical support where they later sent out a form to explain the procedures I carried out to initially culture my cells, subculture my cells, the issue I am having with my cell line, and the storage conditions of my cells. I am hoping that they will send a new NBT-ii cell line to me, so as to avert the additional $500.00 cost to repurchase the cell line. However, if I do not hear back from ATCC regarding this, I will purchase a new cell line so as to not postpone my research any further.