Labware

Instruction of use of UpCell

In vitro cell culture

• Please pre-warm the cell culture medium to be used to 37℃ in the incubator.
• When seeding cells, please use the pre-warmed media to ease attachment of cells.
• UpCell 's temperature-responsive polymer coating is covalently immobilized on the flat surfaces of each cell culture area. The surface becomes slightly hydrophobic above 32℃, allowing adhesion of seeded cells. Therefore please culture cells above this temperature. (Our recommended cell culturing temperature is 37℃.)
• Media change can be performed as with non-polymer coated conventional cell cultureware.
• Attachment time for cells may vary depending on cells.
• To inhibit undesired cell detachment during media changes, please pre-warm culture media to 37℃ prior to changing the media. Observations under a microscope should also be kept as short as possible to prevent cooling.
• Should you have difficulty in cell attachment, surface pre-coating of the surface with collagen, fibronectin, polylysine or use of serum media may improve cell attachment.

Detachment of cultured cells

• UpCell TM's surfaces becomes super hydrophilic below 32℃, prompting cell-surface detachment.
• When harvesting cultured cells, place "UpCell TM" into an incubator set at 20℃ for 30 minutes.
• Alternatively, "UpCell TM" may be set at room temperature (25℃) under sterile conditions to cool.
• Depending on the cell type, cultured cells should detach spontaneously within a short time.
• Cells failing to detach automatically can be detached by gentle plate agitation using a pipet filled with culture media.
• For the preparation of intact single cell suspension by UpCell, it is highly recommended to recover cells before cells reach over confluent and form tight cell-cell junction.
• For the preparation of intact single cell suspension by UpCell, it is highly recommended to recover cells before cells reach over confluent and form tight cell-cell junction.
• The "UpCell" product is intended for single-use only. Please do not re-use the product as the residual matrix and media components may absorb to the surface, comprising performance.
• Store at room temperature. Avoid direct sunlight.
• Sterilization conditions may also comprise cultureware properties.

Cell-sheet harvest using membrane

* The following photo uses doughnut shaped PVDF membrane

1. Remove most but not all of culture medium to prevent cells from drying from UpCell cultureware and leave at room temperature (25C) for 10-30 minutes. Please make sure that your cells are cultured beyond over-confluency.
2. Gently place hygroscopic membrane provided in the product box or PVDF membrane on cells as a "cell-sheet carrier".
3. Place the edge of cells onto the membrane by using sharp forceps and gently peel the membrane with the cell-sheet off from the surface of UpCell.

*1 : Please make sure to place the membrane on cells before reducing the temperature if your cells have weak adherent property (e.g. cells that detaches in less than 10 minutes).