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:-
RPMI + FCS (10%)
+ TPA (200 nM).
[some mouse melanocyte lines including melan-a]
RPMI + FCS (10%)
+ TPA (200 nM) + CT (200 pM).
[primary mouse melanocytes and some mouse melanocyte lines]
Ham's F10 + FCS
(20%) + TPA (48 nM) + IBMX (0.1 mM) at 33o.
[DMEL lines from S. Prince with TS SV40 large T]
RPMI + FCS (10%)
+ TPA (200 nM) + CT (200 pM) + 2ME (100 µM).
[some mouse melanocyte lines]
RPMI + FCS (10%)
+ TPA (200 nM) + 2ME (100 µM).
[some mouse melanocyte lines including melan-c]
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.
RPMI + FCS (10%)
+ TPA (200 nM) + CT (200 pM) + 2ME (100 µM)
+ L-tyrosine (2 mM).
[p-mutant mouse melanocyte lines]
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]
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]
RPMI + FCS (10%)
+ TPA (200 nM) + CT (200 pM) + 2ME (100 µM).
[some mouse melanocyte lines]
RPMI + NCS (10%)
+ TPA (2 nM).
[neural crest-like mouse stem cell lines]
RPMI + FCS (5%).
[mouse melanoma lines]
Further information on all media at top of page.
For melanocyte culture methods see Materials & Methods
RPMI + FCS (10%).
[human metastatic & some primary melanoma lines]
DMEM + FCS (10%).
[all fibroblasts except STO; Rama rat myoepithelial cells and XB2 mouse keratinocytes]
DMEM + FCS (10%)
+ 2ME (100 µM)
on gelatin*.
[P19 mouse EC cells]
1:1 of Ham's
F10:DMEM + 15% HS + 3% chick embryo extract on gelatin*.
[MM14 mouse myoblasts]
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.
Bennett-Sviderskaya Lab - Materials and Methods
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