CellSeed Inc.:Labware

Cell culture condition:
1) Cell type: NIH/3T3, ATCC CCL-1658
2) Cell density: 5.0 x 10⁴ cells/UpCell 3.5cm
3) Culture period: 7 days (over confluent)
4) Media: 10% FBS-DMEM
* Cell culture condition may vary by the cell type.
(ATCC: American Type Culture Collection)

Methodology
1) Seed 5.0 x 10⁴ cells of NIH/3T3 on UpCell 3.5cm and culture for 7 days until cells become over- confluent. (Please note that some cells are difficult to culture beyond over confluency, in which case optimization of the culture condition is required.)
2) Take UpCell out of incubator and remove culture media.
3) Immediately add 50uL of culture media to prevent cells from drying out.
4) Gently place membrane on the cell-sheet. Make sure there is no air bubble in between membrane and cell-sheet.
5) Leave UpCell for 5-6 min at 20-25C.
6) Gently peel membrane off from UpCell using forceps.
7) Place cell-sheet with membrane to the host and wait 1 min. (In the left side of the diagram, the cell-sheet is transferred to sticky gel, mimicking the organ.)
8) Add 1mL of culture medium on the membrane to weaken the hygroscopicity of the membrane to release cell-sheet therefrom.
9) Carefully remove membrane only from cell-sheet using forceps. Make sure the cell-sheet is firmly attached to the host.
10) (Photo) NIH/3T3 cell-sheet has successfully been transferred to the gel. No cells failed to detach from UpCell.
* Cell culture/detachment conditions may vary by the cell types. Optimization may be necessary.