How to Prepare and Plate Semi-Solid Methylcellulose Medium for Cell Culture
Methylcellulose medium is a type of semi-solid medium that can be used for culturing hematopoietic stem and progenitor cells in colony-forming assays, or culturing hybridomas and Chinese hamster ovarian cells for cell line development. This protocol provides instructions for preparing cultureware with semi-solid methylcellulose medium for downstream cell culture.
- Semi-solid methylcellulose medium, such as MethoCult™ or ClonaCell™, must be thoroughly mixed prior to use to ensure a homogeneous mixture and best results for your assay. To mix, shake the bottle vigorously for at least one minute. You should see the medium become opaque with bubbles.
Note: If required, add cytokines or other desired supplements first, and then shake thoroughly.
- Wait 10 - 15 minutes for the bubbles to dissipate.
- Aliquot the medium into working volumes using a blunt-end needle and syringe.
Note: Do not use a pipette to aliquot methylcellulose medium. Methylcellulose medium is viscous and will stick to the inside of the pipette, causing the dispensed volume to be inaccurate.
- Add the appropriate volume of concentrated single-cell suspension.
- Mix by shaking vigorously for at least one minute.
Note: For smaller aliquots, mix by vortexing for ~4 seconds.
- Wait for the bubbles to dissipate.
- Plate the sample using a blunt-end needle and syringe.