Detachment of cells from the UpCell Surface occurs when the temperature of the culture is below 32°C. To avoid undesired cell detachment during handling, the following should be noted:

• Use pre-warmed medium when changing the culture medium;
• Observations under a microscope should be kept short; and
• The use of a hot plate enables you to work with the culture at room temperature for a longer period of time.

Yes, the hydrophilicity of the UpCell Surface is constant below 32°C. It is recommended to perform the detachment in a temperature-controlled, sterile environment. The detachment time depends on the time it takes for the culture to be cooled down to less than 32°C, as well as the cell type used.

It is essential to use a supporting membrane when collecting the cells as a cell sheet from the UpCell Surface. However, you can use a hygroscopic PVDF membrane as an alternative to the supporting membrane supplied with the product (Cat. Nos. 174901 and 174904). Please note that the optimal protocol may vary, depending on the type of membrane used.

The protocol for the detachment of cell sheets may need to be optimized for the cell type that you are using, particularly the degree of confluence at harvest, and the detachment conditions (temperature and incubation times).

Coating of the surface with biomolecules, for example, collagen, fibronectin, poly-lysine and laminin, is possible and should be applied if you experience difficulties in attaching your cells to the UpCell Surface. The coating procedure should be performed at 37°C.

You can use the UpCell Surface with many cell types, including fibroblasts, mesenchymal stem cells, myocytes, keratinocytes, macrophages and different epithelium and tumor cells. If cells do not detach spontaneously from the UpCell Surface by lowering of the temperature to below 32°C, the cells can be detached by flushing the surface with medium or by tapping on the side of the dish or plate. Cells that produce a lot of extracellular matrix and attach very strongly to the surface may be difficult to detach by temperature reduction alone. For these cell types, the simultaneous use of diluted trypsin is recommended.