Brain Tumor Stem Cells by Sheila K. Singh & Chitra Venugopal

Author:Sheila K. Singh & Chitra Venugopal
Language: eng
Format: epub
ISBN: 9781493988051
Publisher: Springer New York

3.Remove the supernatant and resuspend the cells in 2 mL cold Ca2+/Mg2+—free D-PBS solution. Gently pipette to dissociate the spheres.

4.Incubate the cells in cold Ca2+/Mg2+—free D-PBS solution at room temperature for 10 min, while gently pipetting every 2 min.

5.Add a 1:1 (v:v) ratio of HBTC 3160 cells to trypan blue solution.

6.Count the number of live cells using a hemocytometer (see Note 5 ).

7.Spin the cells and resuspend in cold stain buffer to get the desired concentrations depending on the surface antigens to be tested (see Note 6 ).

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