- Suspension cells
Collect cells by centrifuge 5 min at 1100 rpm, remove supernatant and dilute cells to concentration 1x106/ml in fresh medium. Cells can be delivered immediately to GARP core with a scheduled appointment.
- Adherent cells
Adherent cells can be removed by enzymatic digestion using 1 ml of Trypsin 0.25% or 0.05% and incubate in 1 minute at RT or longer depending on your cell type to ensure that cells are unclumped and surface glycoproteins are not degraded, which would impair cell adsorption to ConA Beads.
After detaching adherent cells, add 4 ml of fresh medium to 1 ml of trypsinized cell and centrifuge 5 min at 1100 rpm. Remove supernatant and re-suspend cells in fresh medium.
Dilute cells to concentration 1x106/ml in fresh medium. Cells can be delivered immediately to GARP core with a scheduled appointment.
- Cryopreservation cells
Cells should be frozen under conditions that minimize lysis. Supplement cell culture media with 10% DMSO in media, dilute cells to concentration 1x106/ml, put 1 ml cell in cryovial and slow freeze (-1oC per minute) in an isopropanol-filled container placed at -80oC, using a device such as 'Mr. Frosty' or an equivalent.
Note: Depending on your cell types, cryopreserved conditions can be optimized to ensure that cell viability still maintain >95% after one week stored at -80oC.
Contact GARP core at email@example.com for more information
Cell viability must be > 95%
When providing cells for your CUT&RUN projects, include an additional 30% for quality control checks.
- Live cells should be delivered in fresh medium, kept on ice with a scheduled appointment.
- Cryopreserved cells must be transported in dry ice.