Generating and Maintaining Slice Cultures from Human Glioblastoma Tissue
Transcription
Take human glioblastoma tissue, a primary brain tumor originating from glial cells.
Wash the tissue using an ice-cold processing medium to remove residual blood.
Cut the tissue into pieces and insert it into an embedding mold containing molten agarose.
Chill the mold, allowing the agarose to polymerize and form a stable gel matrix.
Remove the tissue block from the mold and fix it on the specimen plate of a vibratome.
Add the ice-cold processing medium to submerge the block. Pass an oxygenated gas mixture to aerate the medium, maintaining tissue viability.
Section the embedded tumor tissue into thin slices.
Transfer a slice into a petri dish containing the ice-cold processing medium to preserve the tissue architecture.
Then, plate the tissue slice on a permeable membrane insert in a well containing a slice culture maintenance medium.
Maintain the slice in physiological conditions. The slice culture is ready for downstream assays.
