Temperature-responsive cell cutureware for cell collection
Temperature responsive polymer is fixed to the surface of the cultureware at nano-thickness level.
Cells are cultured at 37 degrees Celsius and collectable approximately 10-30min after controlling temeperature to 20-25 degrees Celsius.
3mmx3mm "Grid-walls" are provided on the surface (for dish configurations only) for enhancement of collection of cells in a form of single cells or small colonies instead of contiguous cell-sheets.
"Grid-wall" is an optimized gouge on the surface of the dish which inhibits cells from growing over the grid.
- 6cm and 10cm dish type is now available!
- 3mm × mm "Grid walls" are provided on the surface (for dish configurations only) for enhancement of collection of cells in a form of single cells or small colonies.
Relative advantages of RepCell™ (UpCell® with Grid)
- Collection of intact viable cells without enzymatic processing.
Trypsin is well known for its deterioration to the functions of cells. Maintenance of their activities is possible when RepCell™ is used for cell culture.
- Harvest cells by mere reduction of temperature.
It completely abandoned the usage of trypsin when collecting cells since cell detachs by just lowering the temperature. Saves time for laborious pipetting procedure. Our internal report suggests that total labor spent on these procedures were cut down to 1/5. 96 multi-well configuration is especially suitable for cell cloning by limited dilution.
- Suitable cell type for RepCell™ (UpCell® with Grid)
- Ideal for harvesting fragile cells such as stem cells and dendritic cells. Better results can be expected when preparing cells to be used for cell therapy.
- Collection of intact and viable single cell suspension.
- For cell screening.
- For research on extra cellular matrix.
RepCell™ is suitable for culturing cells shown in the following diagram.
* By adjusting the thickness of the grafted polymer, those cells difficult to attach or detach with the standard RepCell™ will become attachable or detachable by mere reduction of temperature.
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
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Application data / References
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