To perform a primary culture of non-parenchymal liver cells from mice, it is essential to choose the appropriate medium, determine the percentage of fetal bovine serum (FBS) to use, and decide on the necessary supplements. It is important to note that specific protocols may vary based on the intended application and the preferences of your laboratory.

For the medium, a commonly used choice for primary cell cultures is Dulbecco's Modified Eagle Medium (DMEM) or RPMI-1640. Both media are widely available and suitable for the growth of liver cells. The selection of the medium may depend on the specific requirements of your experiment or the protocols followed by your research group.

Regarding the percentage of FBS, a common range is between 5% to 10%. The choice of the exact percentage depends on the specific cell type and experimental conditions. It is advisable to optimize the FBS concentration based on the viability and growth characteristics of the non-parenchymal liver cells in your particular experiment.

As for supplements, commonly added components include L-glutamine, penicillin-streptomycin (antibiotics), and non-essential amino acids. The recommended concentration of L-glutamine is typically 2 mM, while the antibiotics are generally added at concentrations of 100 units/mL of penicillin and 100 μg/mL of streptomycin. Non-essential amino acids are often added at a final concentration of 1% or as specified by the supplier.

Additionally, considering the variability in experimental conditions, it is recommended to perform optimization experiments to ensure the optimal culture conditions for your non-parenchymal liver cells.

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