This is a product Nunc/ThermoFisher. The manufacturer offers the same detachment information, regardless of dish size or number of wells.
The following example protocol for cell suspensions:
1. Mouse Peritoneal Macrophages were washed with Ca2+– and Mg2+–free PBS to remove non-adherant cells
2. Fresh medium including FBS was added to the plate.
3. Cultures were incubated for 30 minutes at 20°C prior to harvesting.
The following example protocol for harvesting cells as a single sheet:
Madin-Darby Canine Kidney (MDCK) cells in Dulbecco’s Modifi ed Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), penicillin (100 units/mL) and streptomycin (100 μg/mL) were seeded at 5.7 x 104 cell/cm2 in a 3.5 cm dish with UpCell Surface. The cells were incubated for 21 days at 37°C in a humidifi ed atmosphere of 5% CO2 in air. The cells were then harvested as an intact sheet:
1. The dish was placed at 20°C for 60 min.
2. A supporting membrane was placed on top of the cell layer and the culture medium was aspirated
3. The edge of the membrane was gently released from the dish using forceps, and the membrane with the attached cell layer was then transferred to a cell culture insert and incubated at 20°C for 30 min.
4. Sufficient culture medium was added to cover the cell sheet and membrane and the membrane was removed from the cell layer using forceps
