PhenoDrive-I

• Dissolve the contents of each vial in ethanol or any aqueous medium.
• Dilute the substrate powder to a concentration of 0.01mg/mL to 0.1mg/mL in any sterile buffer solution pH 7.4
or in 75% ethanol (for rapid coating) and filter.

Coating of 96 and 24 well plates

• Under laminar flow pipette the reconstituted solution into each well (50 µL for 96 wells, 200 µL for 24 wells)
• Allow cast coating by solvent evaporation under sterile conditions (UV irradiation is recommended). Time for evaporation will vary depending on substrate, concentration and/or volume (please contact technical support for assistance if required) and wash once dried.
• SERUM-FREE SEEDING OF CELLS IS RECOMMENDED. Serum can be added after cell adhesion (e.g. 3 hours after seeding).

Coating of 3D polymer scaffolds

• Pipette a sufficient volume to ensure complete
 coverage of the scaffold. The required volume may change depending on the scaffolds size, porosity, chemical composition and swelling properties. Scaffolds may experience temporary increased swelling when ethanol solutions are used. Equilibration in tissue culture media should restore their original degree of swelling
• Equilibration in tissue culture media should take into account diffusional constraints related to the scaffolds physicochemical properties. This is particularly important if an ethanol solution is used as it could result in cell cytotoxicity 


• Solubilise the PhenoDrive in SERUM-FREE tissue culture medium specific for the cell type to be cultured by reconstituting the vial powder as described in the powder reconstitution protocol.
• Mix the resulting solution with cell suspension to reach a final concentration ranging from 0.001% to 1% (v/v). A typical mixture with cell suspension varies from 40,000 cells/mL to 1,000,000 cells/mL and incubate for 20 min at room temperature or 37 °C under gentle rotary conditions.
• Seed the cells as normal. It is recommended that the seeding of PhenoDrive-driven cell constructs is further supported by prior coating of the surface with the same PhenoDrive formulation as outlined in the coating procedure described for 2D culturing conditions.

PhenoDrive is supplied as a lyophilised powder for easy reconstitution in a range of aqueous or polar solvents. The lyophilised powder can be stored for up to 6 months at 4°C. Reconstituted PhenoDrive can be stored at -20°C and used within 3 months of freezing.

Each vial of PhenoDrive contains 1 mg of freeze-dried product which when reconstituted allows the coating of a 96-well or 24-well plate