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Cell Culture Media

To find the medium for a particular cell line, please start from the Cell Bank holdings page, and click on the medium number listed for that cell line.
For melanocyte culture methods see Materials & Methods.

All cultures are incubated at 37°C with 10% v/v CO2. This give pH 6.9 - 7.0 with RPMI 1640 and Ham's, or pH 7.4 with standard DMEM.
It is not recommended to use 5% CO2 with either medium - too alkaline. However if there is no other option, a medium can be acidified with HCl for use with 5% CO2 - see protocol.

RPMI 1640 ("RPMI") is supplemented with penicillin (100 units/ml), streptomycin (100 µg/ml), glutamine (2 mM), and additional phenol red (7.5 µg/ml), for easier monitoring of pH. See Materials for more details if required.

DMEM is supplemented with penicillin (100 units/ml), streptomycin (100 µg/ml) and glutamine (4 mM).

Abbreviations for components:
(details of sources etc in Materials listing)

2-ME - 2-mercaptoethanol
CT - cholera toxin. (Caution: toxic, handle with care and treat media with bleach before disposal.)
EDN1 - endothelin 1
FCS - foetal calf serum
FGF2 - fibroblast growth factor 2
HS - horse serum
hSCF - human stem cell factor
IBMX - 3-isobutyl-1-methylxanthine
mSCF - mouse stem cell factor
NCS - newborn calf serum
PTU - phenylthiourea (also known as phenylthiocarbamide) (Caution: toxic.)
RPMI - RPMI 1640
TPA - 12-0-tetradecanoyl phorbol acetate. TPA is hazardous - see Materials page for how to handle.

* gelatin is disolved in water (100 µg/ml) and used to pre-treat dishes.

For melanocyte culture methods see Materials & Methods

Medium listings:-

  1. RPMI + FCS (10%) + TPA (200 nM).
    [some mouse melanocyte lines including melan-a]

  2. RPMI + FCS (10%) + TPA (200 nM) + CT (200 pM).
    [primary mouse melanocytes and some mouse melanocyte lines]

  3. Ham's F10 + FCS (20%) + TPA (48 nM) + IBMX (0.1 mM) at 33o.
    [DMEL lines from S. Prince with TS SV40 large T]

  4. RPMI + FCS (10%) + TPA (200 nM) + CT (200 pM) + 2ME (100 µM).
    [some mouse melanocyte lines]

  5. RPMI + FCS (10%) + TPA (200 nM) + 2ME (100 µM).
    [some mouse melanocyte lines including melan-c]

  6. RPMI + FCS (10%) + TPA (200 nM) + 2ME (100 µM) + PTU (200 µM).
    [some mouse melanocyte lines with abnormally high pigmentation]

    Further information on all media at top of page.
    For melanocyte culture methods see Materials & Methods.

  7. RPMI + FCS (10%) + TPA (200 nM) + CT (200 pM) + 2ME (100 µM) + L-tyrosine (2 mM).
    [p-mutant mouse melanocyte lines]

  8. RPMI + FCS (10%) + TPA (200 nM) + CT (200 pM) + hSCF (10 ng/ml) + EDN1 (10 nM).
    [normal and immortal human melanocytes, new (SGM) RGP melanoma lines].
    8a: Same medium except 20 nM TPA, not 200 nM
    [New VGP melanoma line SGM5]

  9. RPMI + FCS (10%) + FGF2 (40 pM) + mSCF (20 ng/ml).
    (Earlier passages in RPMI + FCS (10%) + TPA(2 nM) + FGF2 (40 pM) on XB2-feeders).
    [mouse melanoblast lines]

  10. RPMI + FCS (10%) + TPA (200 nM) + CT (200 pM) + 2ME (100 µM).
    [some mouse melanocyte lines]

  11. RPMI + NCS (10%) + TPA (2 nM).
    [neural crest-like mouse stem cell lines]

  12. RPMI + FCS (5%).
    [mouse melanoma lines]

    Further information on all media at top of page.
    For melanocyte culture methods see Materials & Methods

  13. RPMI + FCS (10%).
    [human metastatic & some primary melanoma lines]

  14. DMEM + FCS (10%).
    [all fibroblasts except STO; Rama rat myoepithelial cells and XB2 mouse keratinocytes]

  15. DMEM + FCS (10%) + 2ME (100 µM) on gelatin*.
    [P19 mouse EC cells]

  16. DMEM + FCS (10%) on gelatin*.
    [C2 mouse myoblasts]

  17. 1:1 of Ham's F10:DMEM + 15% HS + 3% chick embryo extract on gelatin*.
    [MM14 mouse myoblasts]

  18. DMEM + FCS (5%).
    [RT4 rat pluripotent neuroblastoma lines]

  19. DMEM + FCS (10%) + hydrocortisone (50 ng/ml) + insulin (50 ng/ml).
    [Rama rat mammary glandular epithelial and mammary tumour lines]


    Further information on all media at top of page.
    For melanocyte culture methods see Materials & Methods.

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