JoVE Encyclopedie van Experimenten
Kankeronderzoek
0 weergaven • 2:57 min. • April 30th, 2023
- To begin, add trypsin to a plate containing leukemia cells co-cultured with bone marrow stromal cells for cell dissociation. Centrifuge and re-suspend the pellet using pre-warmed media to maintain cell viability. The G-10 column contains cross-linked dextran beads that swell in water to form a gel matrix.
Equilibrate the G-10 column with pre-warmed media to fill the space between beads, ensuring slow elusion of the media from the column. Maintain a suitable temperature in the column to prevent the formation of air bubbles. Load the cell mixture slowly onto the gel beads in the G-10 column. Incubate the cells on the G-10 column at room temperature for the desired duration. Based on cell surface characteristics and size, the gel beads preferentially restrict the larger bone marrow stromal cells, allowing the separation of the leukemia cells.
Add pre-warmed media to the column and collect the suspension of leukemia cells. Centrifuge the cell suspension and resuspend the pellet in an appropriate buffer. Use the pure population of leukemia cells for further analysis. In the following example, we will separate a pure population of leukemia cells from stromal co-culture using the G-10 c
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