Cell adhesion was measured as a function of the mechanical force required to detach cells from a cultured plastic surface. RBL-2H3 cells were plated at 3 × 105 cells per well in a plastic six-well plate in supplemented MEM and incubated for 24 hours. Medium containing any nonadherent cells was removed, and the adherent cells were washed with 1 ml of PBS. Cells were then covered with 2 ml of PBS and placed onto a benchtop orbital shaker at varying speeds for 30 min at room temperature. After shaking, the number of detached cells in solution was counted using a hemocytometer. The remaining adherent cells were detached by incubation with trypsin and EDTA for 10 min and counted in the same manner. Spontaneous growth in suspension was determined by plating the RBL-2H3 cells at 3 × 105 cells per well in a plastic six-well plate in supplemented MEM and incubating them for 24 to 72 hours before assessing the suspension and adherent fractions as described earlier. Cell dendricity was determined by collecting differential interference contrast (DiC) images of RBL-2H3 cell populations cultured on glass coverslips for 24 hours after plating at 3 × 105 cells/ml. Cells were qualitatively classed as dendritic if they exhibited protruding dendrites from the main cell body, whereas those cells with roughly spherical morphologies were classed as nondendritic.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.