Thursday, 5 September 2013

Training in the Lab - Thawing the Cryopreserved Cells and Establishing a Culture

Over the past few days at XCellR8, I have been training and learning how the Balb/c3T3 Cells are cultured. 

Bushra started by showing my step-by-step how each stage of the cell-culture process is carried out. 
Firstly the cryopreserved cells are removed from the liquid nitrogen and then rapidly thawed in the water bath.


During the demonstration I learnt about important aseptic techniques, such as ensuring the ampoule is not fully immersed in the water bath, as this could increase the risk of contamination.


After the cells have been thawed, the contents of the ampoule is centrifuged to separate the cells from the supernatant. I learnt about how to counterbalance the centrifuge and the importance of adding culture medium to the cell suspension to ensure the cells survive!


After the cells have been centrifuged, they are resuspended in medium, and a cell count is performed. A cell count is performed using a haemocytometer and trypan blue exclusion stain. This part looked quite confusing and I think it will take some practice before I perfect it!



After the concentration of cells has been determined, the cells are seeded at the recommended density in a T25 flask. The flask is then placed in the incubator which provides the perfect conditions for the cells to grow.


I am looking forward to revisiting the cells over the next few days and seeing how they grow!
Lottie 



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