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Monday 5-24: Today was the type two cell extraction procedure. I am grateful to Steve for helping me to understand the steps involved and showing me the right way to do this experiment. The expected yield is between 30 million and 45 million. The procedure as I understand it currently can be viewed by clicking here.
Also checked the number of T2I cells present and determined it was enough to start coating plates. I used 10 60 mm plates with 2E5 cells each. After 96 hours, the medium containing all the growth factors will be taken from these cells.
T2I cells stick to the plastic of their growth containers. Therefore, to extract and spin down these cells, they must be washed twice with PBS followed by a 2 minte incubation in Trypsin solution. The trypsin must then be neutralized with DMEM.
Tuesday 5-25: Met with Go-Shau to discuss experimental protocol and procedures.
Wed: Performed experimental BAL for macrophage yield--6 million
Thu: Solutions for next week.
Fri: Extra reading for presentation next week.
Go to week 4--the exciting experiment week!!!
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