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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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