Biogelx gels are supplied as a lyophilized powder (Biogelx Powder), which is rehydrated with water to prepare a Pre-Gel solution of the user’s desired concentration. This Pre-Gel is added to a well plate, along with cell culture media which promotes gelation. Cells can be cultured inside the gel (3D culture) or on top (2D culture).
Gelation is initiated by cell culture media and/or salt-containing buffers. The material will remain in the pre state until media and/or salt-containing buffers is added.
When rehydrating the Biogelx powder, it is possible to prepare a higher concentration Pre-Gel stock solution, which can be diluted to Pre-Gels of lower concentrations. However, dilutions should only be carried out at the time of Pre-Gel preparation and not in the future. This method may be employed, if the user wishes to test a wide range of final gel stiffnesses.
Pre-Gel solutions of higher concentration can be highly viscous. If the transfer of highly viscous Pre-Gels proves difficult, the use of a wide orifice pipette tips, or a pipette tip with 1 cm cut from the end is recommended.
If any air bubbles are present in the Pre-Gel solution, remove these by placing the solution in a bath sonicator for 10 seconds or by centrifugation.
When mixing cells with the Pre-Gel solution (for 3D culture) ensure the pipette tip does not leave the Pre- Gel when mixing, as this can introduce air bubbles into the gel structure.
First addition of media to the Pre-Gel solution should be performed CAREFULLY; gently pipetting cell media dropwise onto the centre of the Pre-Gel. Pipetting down the side of the well can disrupt the material.
Lower concentration Pre-Gel solutions will have lower viscosities and may appear almost water-like in a vial. However, when added to a well/insert the surface tension of the Pre-Gel ensures that it is not disrupted by/mixed with the media.
It is necessary to handle the material very carefully when performing media changes. Do not use a vacuum aspirator to remove media from above the hydrogel. Avoid direct contact with the hydrogel.