PhenoDrive-R facilitates cell attachment and is based on the modulation of a protien tripeptide sequence present in the extracellular matrix and extracellular matrix proteins.
Why do I need PhenoDrive-R?
PhenoDrive-R has been used successfully in culture with mesenchymal stem cells, cancer cell lines, epithelial cells and osteoblasts. Beta tests using PhenoDrive-R have shown an increased spheroid formation combined with stem cell marker expression.
Frequently Asked Questions
How do I prepare my PhenoDrive powder?
- 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.
How do I prepare my PhenoDrive for cell culture?
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).
How do I prepare my PhenoDrive powder for scaffold coating?
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
How do I prepare my PhenoDrive for use in suspension?
- 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.
How do I store my PhenoDrive?
PhenoDrive-I 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-I can be stored at -20°C and used within 3 months of freezing.
How many wells does a 1mg vial coat?
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
