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The MRC-9 cell line is a human diploid fibroblast strain derived from the lungs of a White female fetus at 15 weeks gestation. It was established for long-term use in biological research, vaccine production, and diagnostic virology. The cells exhibit adherent growth properties and maintain a stable, diploid karyotype throughout serial propagation. Importantly, MRC-9 cells possess normal chromosomal features and are non-tumorigenic under typical conditions. However, they have demonstrated tumorigenic properties when tested in semisolid media, but not when implanted in immunosuppressed mice. MRC-9 cells have been widely used for viral susceptibility studies, showing broad susceptibility to various viruses such as poliovirus, adenovirus, rubella, and cytomegalovirus. These cells are particularly valuable for their use in virology research and vaccine development. While their proliferation rate is slower compared to other human fibroblast strains like MRC-5, MRC-9 cells can still undergo a significant number of population doublings, with reports of up to 46 doublings.
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- cultureMedium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
- supplements: Supplement the medium with 10% FBS and 1% NEAA
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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