A matrix plug can remain in vivo for at least one week.

Use ice-cold serum-free culture medium or PBS at pH 7.4.

Use a pre-cooled pipette or syringe, and pre-cool pipettes and tips as well. Do not touch the bottom of the bottle while aspirating. Dispense slowly and avoid excessive force. When using a serological pipette to dispense 5 mL, aspirate 6 mL and stop dispensing when 1 mL remains in the pipette. When using an automatic pipettor (Pipetman), press to the second stop to aspirate and dispense only to the first stop.

The higher the protein concentration, the more viscous the matrix. At concentrations above 13.0 mg/mL, the matrix may appear very thick. Undiluted GelNest™ Basement Membrane Matrix products are generally viscous. A high-concentration matrix can be used directly without dilution, for example when culturing tumor cells and/or angiogenic factors. After injection into mice, the cells remain localized, facilitating in situ analysis and/or subsequent excision. Alternatively, the matrix can be diluted and used in the same manner as a standard-concentration product; the specific dilution should be determined according to experimental requirements.

In addition to protein concentration, viscosity may be affected by temperature fluctuations during transportation and by storage conditions. Dry ice must be used throughout transportation. If the storage freezer has an automatic defrost function, the temperature increase during defrosting may cause the matrix to gel. Therefore, do not store GelNest™ Basement Membrane Matrix in this type of freezer. Minimize freeze-thaw cycles, aliquot newly received matrix into single-use portions, and keep the matrix on ice during every thawing procedure.

First, check whether the cell seeding density is too high. The volume of GelNest™ Basement Membrane Matrix should be equivalent to the volume of culture medium in the cell culture system. If the matrix is diluted to an excessively low concentration, the resulting gel may readily detach from the surface of the tissue-culture vessel.