There is no significant difference in the percentage of cells that will differentiate into Dendritic Cells. But when using cryopreserved cells the initial cell loss will be higher compared to when fresh cells are used. i.e. the final number of differentiated cells that can be expected will be higher with fresh cells as a starting material due to lower cell death rate.

You should use complete DC Generation Medium/DC Generation Medium XF (with all the cytokines). As cells are metabolically active media should be changed every 3 days. We have observed that the dendritic cell phenotype remains stable for up to 7 days.

According to our experience; they can be maintained for 3 to a maximum of 7 days. However; the morphology will change and they will look more and more "degenerate".

Usually we do not recommend specific plasticware but we have found that with PromoCell’s Dendritic Cell and Macrophage Generation Media choice of plasticware can have a lot of an influence. For our Dendritic Cell Generation Media we recommend to use tissue culture vessels from BD Falcon^TM.

The PromoCell DC Generation Media have been developed for the easy and efficient generation of immature as well as fully mature myeloid dendritic cells from peripheral blood monocytes. For freshly isolated mononuclear cells (MNC) and monocytes we recommend our DC Generation Medium XF (C-28052); for cryopreserved monocytes our DC Generation Medium (C-28050). When using DC Generation Medium (C-28050) with fresh MNC; the Monocyte Attachment Medium (C-28051) is needed in a first step for efficient adherence of the monocyte fraction. Please see Instruction Manual; Application Note and the graph below for further details.

Our DC Generation Medium XF (C-28052) has a xeno-free formulation. It provides a complete media system (ready-to-use all cytokines included) and shows efficient and reproducible in vitro maturation of moDCs from freshly isolated peripheral blood monocytes.

It takes 7-8 days to generate fully mature myeloid dendritic cells.

DC Base Medium XF (C-28054) consists of 250 ml Basal Medium + SupplementMix. It does not include cytokines and must therefore be supplemented according to the user's individual needs.

DC Generation Medium XF (C-28052) is ready-to-use and consists of 250 ml Basal Medium + SupplementMix + appropriate cytokines.

Both; Base Medium XF and Generation Medium XF have a xeno-free formulation.

Yes the specialized PromoCell media already contain the optimal amount of L-glutamine. Please don't add extra L-glutamine as this can be toxic for the cells.

A concentration > 10% FBS is needed to completely inactivate the trypsin. As most of PromoCell's growth media are serum-reduced or serum-free; the use of a trypsin inhibitor like TNS is highly recommended.

If antibiotics are deemed necessary the recommended final concentrations are: 100 U/ml penicillin + 100 µg/ml streptomycin or 50 µg/ml gentamicin + 50 ng/ml amphotericin B Please note: Addition of antibiotics can reduce the growth rate of the cells.

After addition of the SupplementMix or SupplementPack to our basal medium do I have to add FCS you obtain the complete growth medium. No further supplementation with serum or growth factors is required. Please note: Our media do not contain antibiotics. If you wish to use antibiotics you can add penicillin/streptomycin or gentamicin/amphotericin B at standard concentrations. Addition of antibiotics can however reduce the doubling time of the cells up to 30-40%.

Yes you can aliquot the SupplementMix upon delivery and freeze down 2 or 4 individual aliquots at -20°C. This way you can prepare smaller volumes (2 x 250 ml or 4 x 125 ml) of complete culture medium and thus extend the time you can use the medium.

Upon arrival the basal media should be stored between 4°C and 8°C the supplements at -20°C.

Please do not freeze our cell culture media. Freezing can lead to irreversible precipitation of media components and the quality can no longer be assured.

The qualitative and quantitative composition of the supplements can be found on our website and in the data sheets of the specialized media. When there is no such information specified; the composition of the supplements is confidential.

The formulation of our basal media is proprietary information. If you need to know the concentration of a particular component for your experiments please contact the PromoCell Technical Customer Service.

It has been shown that phenol red has estrogenic properties. Phenol red-free media are therefore generally used in studies evaluating steroid hormone action in cultured estrogen-responsive cells (Berthois et al. 1986). Furthermore phenol red can also interfere with some analytical methods like photometric analyses.

The Growth Medium Kit allows customization of the end concentrations of growth supplements. It is therefore more flexible than the Medium "ready-to-use" and can eg. be used to prepare a starvation medium. Please note: Modification of supplement concentrations may have an impact on cell growth. You should test in advance whether and for how long the cells can survive the altered culture conditions.

PromoCell Cell Culture Media "ready-to-use" consist of basal medium and SupplementMix. PromoCell Culture Media Kits consist of basal medium and SupplementPack. Addition of the supplements (SupplementMix or SupplementPack; respectively) to the appropriate basal medium will result in identical growth media.

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Dendritic Cell Generation Medium XF
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