Tissue+Culture+Protocol

Tissue Culture Protocol

First decide dimensions/volumes. See tissue culture volumes and dimensions page or tissue culture volumes and dimensions from Danielle page


 * Wash cells with ___mL of 1X PBS
 * Release PBS on the side of the flask the cells are not on
 * cap the flask, and gently move the flask so that the PBS washes the side with the cells
 * Aspirate away the 1X PBS
 * Detach cells with ___mL of 0.05% Trypsin
 * Release Trypsin on the side of the flask the cells are not on
 * cap the flask, and gently move the flask so that the trypsin goes over the cells
 * Incubate for about 2 minutes
 * Hit the flask so the cells will release
 * may want to check status of cells under the microscope at this point
 * Neutralize the trypsin by adding ___mL of media
 * Release media on the side of the flask the cells are not on
 * cap the flask, and gently move the flask so that all the trypsin is neutralized
 * If passing, first decide what ratio you'd like to do it. Remove all but 1mL of media/cells and add media to the necessary corresponding volume.
 * Put date on front of the flask and what passage number it is. Return to 37C incubator, 5% CO2.

Protocol from the Current Protocols in Cell Biology (2007):

Starting Tissue Culture Cells

Cell Lines