Mononuclear Cell Medium

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FCS is a natural product consisting of many different components like salts hormones vitamins trace elements proteins and enzymes. There can be large lot-to-lot variations between different serum batches regarding the concentrations of growth-promoting factors. The higher the serum content in a culture medium the higher the impact of the variations on the cell culture system. For this reason PromoCell has developed several serum-reduced and serum-free media where part of the serum has been replaced by more defined factors like cytokines hormones or vitamins.Media with reduced serum concentrations have the benefit that they produce more standardized culture conditions over a long time span.

Serum turbidity is usually caused by cryoprecipitation of lipid components during freezing and thawing. The more times the serum is subjected to freeze/thaw cycles the more turbidity is noticed. It can be minimized by freezing the serum in aliquots at -20°C and thawing these aliquots individually at the time of use.

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.

The practice of heat inactivation was originally developed when only serum from adult animals was available. Adult serum contains high serum complement which may destroy cells under certain conditions. Heating serum (30 min 56°C) is intended to inactivate the complement. Today serum is often heat-inactivated without any evidence of beneficial effect. When using FCS (fetal calf serum) heat inactivation is not necessary for most cell lines or cell types. PromoCell does not use heat-inactivated FCS for the preparation of their growth media.

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.

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 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.

Mononuclear cells are mostly used in immunology; infection biology; hematology and cancer research to study subpopulations of blood cells. Our Mononuclear Cell Medium (C-28030) is intended for short-term maintenance (up to 48 hrs) of the thawed hMNC before you proceed with your experiments. The number of PDs will depend on the subsequent cell culture conditions and is not determined by PromoCell. Please note: Depending on the conditions; the ratio of the subpopulations will gradually change; as the different blood cell types behave in different manners. Researchers normally start soon after thawing to either select the cell type of their interest (e.g. hematopoietic cells; endothelial progenitor cells) or perform experiments with all populations of hMNCs (e.g. to study effects on toxicity; viability or metabolism).

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.