https://bridgeslab.sph.umich.edu/protocols/api.php?action=feedcontributions&feedformat=atom&user=NicoleP 2026-04-19T18:20:13Z User contributions MediaWiki 1.45.1 https://bridgeslab.sph.umich.edu/protocols/index.php?title=Splitting_Cells&diff=796 2013-09-16T21:15:21Z

<p>NicoleP: changed serum names</p> <hr /> <div>==Materials==<br /> *Media (FBS or NCS as required): Prepare by filtering together 5 mL PSG, 50 mL COS-FBS and one bottle of DMEM<br /> *PBS -/-<br /> *0.05% Trypsin<br /> <br /> ==Protocol==<br /> #Warm PBS and Media in water bath<br /> #Wash cells twice with 10 mL (per 10 cm dish) PBS -/-<br /> #Add 1 mL trypsin and sit in the hood for 2-5 min<br /> #Add 10 mL media to each new dish<br /> #Check cells for trypsinization, and if necessary tap the cells<br /> #Add 9 mL media to trypsinized cells<br /> #Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X)<br /> #Replace plates in the incubator<br /> <br /> ==Cell Specific Notes==<br /> *3T3-L1 fibroblasts have special considerations regarding confluence. See [[Differentiation of 3T3-L1 Cells]]<br /> *RAW 264.7 cells are scraped, not trypsinized. See [[Culturing RAW 264.7 Cells]]<br /> *S2 cells are grown at 28C without extra CO2. See [[Culturing S2 Cells]]<br /> [[ Category:Cell Culture ]]</div>

NicoleP