Labware

Instructions of usage

Culturing cells

• First of all, pre-warm the medium to 37 degrees celsius in the incubator.
• When seeding cells, please do use the pre-warmed medium to ease attachment of cells.
• Seed cells evenly over the culture surface to prevent cells being concentrated in one area.
• The surface becomes hydrophobic above 32 degrees celsius therefore cells must be cultured above this point.
• Medium change can be carried out as if using normal cultureware without polymer. Please use pre-warmed medium for change.
• When cells are difficult to attach, please use medium that contains FCS.
  (* Attachment time of the cells may vary depending on the type of the cell.)

Detachment of cells

• The surface of the cultureware becomes hydrophilic under 32 degrees celsius. Cells are easier to detach under this temperature.
• When collecting cells, lower the temperature down to approximately 20 degrees celsius and wait about 30 minutes. Depending on the type of the cells cultured, they detach automatically.
  (* Same results can be achieved by leaving the cultureware in the refrigerator set at 4 degrees celsius for 10 minutes.)
• The cells that did not come off automatically can be detached by agitating them by pipet with medium.
  (* Some cells may not detach completely. The results may vary by the type of the cells.)
• Detachment of cells are easier when the temperature is lowered before cells become confluent.
• The product is intended for one time use only. Please do not re-use the product.
• The product is intended for research use only. Any other use is not warranted by CellSeed Inc.

Intact single cell collection with RepCell (UpCell with Grid)

• Seed single cell suspension on to RepCell and incubate for 24 hours to establish matrix production.
• Then harvest by temperature change before proliferation begins again.

	Normal culture 1x104 cells seeded and incubated for 24 hrs. RepCell 6cm dish used	Recovered cell suspensions
NIH/3T3

Passage culture with RepCell

• Culture cells on RepCell and harvest at 70-80% confluency where cell-cell junctions are not established and are readily separated by slight agitation and media pipetting.

	Normal culture 1x105 cells seeded and incubated for 72 hrs. RepCell 6cm dish used	Recovered cell suspensions (After slight pipetting)
NIH/3T3