AlanFung:LabNotes/Cell/2009-1-21

From ZhangLabWiki
Revision as of 01:10, 22 January 2009 by >Alan6017518 (New page: 1.Recovered 1 vial of GM18506B and GM18508B for testing 2.Freezed down 2 vials of GM18507C 3.Keeep the 2 T25 GM18506B and GM18508B at 200,000cells/mL 1. Prepared 2 new T25flask added 19mL...)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

1.Recovered 1 vial of GM18506B and GM18508B for testing 2.Freezed down 2 vials of GM18507C 3.Keeep the 2 T25 GM18506B and GM18508B at 200,000cells/mL

1. Prepared 2 new T25flask added 19mL fresh medium into each labeled flask, thaw 1 vial(1mL) of GM18506B and 1 vial(1mL) GM18508B in 37c waterbath (5million cells/mL) Calculation of cell density 5,000,000cells *1mL/200,000cells =25mL. However we dont want to have more than 20Ml of fresh medium hence resuspend cells in 20mL medium. New concentration= 250,000cels/mL Cells will be checked tomorrow and remove part of the medium to remove DMSO

2. Combined two T25 flasks of GM18507C cells performed cell counting 457,469cels/mL*40mL=18,298,760cells in total. Take 6 million cells for expansion 6,000,000cells *40mL/18,298,760cells = 13.1mL Aspirate 13mL suspension aliquot to two new T25 flask and make up to 15mL each. Spin down remaining cells and resuspend with 2mL medium w/ 5% DMSO aliquot to 2 vials and transfer on ice to -80 freezer.

3. Cell Count GM18506B: 565,459cells/mL*15mL=8,481,885 cells GM18508B: 583,437cells/mL*20mL=11,668,740 cells Both flask dump cell suspension to about 5mL and add in 10mL fresh medium