Cell culture & media
Everything between a thawed vial and a confluent flask: a base formulation matched to your line, serum or a defined supplement to support it, a dissociation reagent to passage it, and, if you're maintaining a stable line, a selection antibiotic to hold the pressure.
Pick a base formulation
Start from the classical formulation your line was validated on, DMEM, DMEM/F-12, RPMI-1640, IMDM, MEM alpha, or a specialty medium for a non-standard host. Check glucose, L-glutamine format (free vs. stable dipeptide), and phenol red before you order.
Supplement it
Add heat-inactivated FBS, horse serum, or a defined serum-free supplement (B-27, N-2, ITS) depending on whether the protocol tolerates serum lot variability or needs a chemically consistent alternative.
Passage the culture
Detach adherent cells with trypsin-EDTA, an Accutase-style or TrypLE-style recombinant protease for gentler dissociation, or a tissue-specific enzyme (collagenase, dispase) for primary isolations.
Hold selection pressure
Stable lines need continuous antibiotic selection, G418, puromycin, blasticidin, or hygromycin for mammalian resistance markers, or gentamicin / amphotericin B to keep primary cultures clean.
DMEM, High Glucose, with L-glutamine
Dulbecco's Phosphate-Buffered Saline (DPBS), no calcium, no magnesium
Endothelial Cell Growth Medium
PBS (1×), with calcium and magnesium
Penicillin-Streptomycin (100×)
RPMI-1640, with L-glutamine
Every product below is pulled live from the catalog, not hand-curated marketing copy.
Serum-containing or serum-free?
FBS carries undefined growth factors and lot-to-lot variability that can shift proliferation and differentiation assays. A defined serum-free supplement (B-27, N-2, ITS) trades some robustness for reproducibility, the standard choice for stem cell and neuronal work where lot variation confounds a phenotype.
With or without phenol red?
Phenol red is a weak estrogen-receptor agonist and fluoresces in some imaging channels. Use phenol-red-free formulations for hormone-sensitive lines, live-cell fluorescence imaging, or any assay reading absorbance/fluorescence in the visible range.
Trypsin-EDTA or a recombinant dissociation reagent?
Porcine trypsin is inexpensive and works for most robust lines, but its animal origin and batch variability are a problem for GMP-track or xeno-free protocols. Accutase-style and TrypLE-style recombinant proteases dissociate more gently (useful for sensitive primary cells and stem cells) and are animal-origin-free.
How much selection antibiotic should I use?
Kill-curve it on untransfected cells first, the effective concentration for G418, puromycin, blasticidin, and hygromycin varies 5–10× across cell lines. Use the lowest concentration that kills 100% of untransfected cells within 5–7 days.
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