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Monocyte Attachment Medium
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Name: Monocyte Attachment Medium
SKU: C-28051

Defined, animal-component free, and protein-free medium for efficient adherence selection of monocytes from freshly isolated human mononuclear cells.

Unit 250 ml

Cat.No.

C-28051

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Our Monocyte Attachment Medium allows for the efficient adherence selection of monocytes from freshly isolated human mononuclear cells while maintaining optimal cell health. Thus, the time-consuming and costly immunomagnetic purification of monocytes is dispensable prior to experiments requiring adherent monocytes as a starting material.

With properly performed washing steps (see Product Manual) large numbers of 80-90% pure monocytes can be obtained with our Monocyte Attachment Medium. Typical applications are generation of dendritic cells/macrophages as well as phagocytosis assays.

Product Manual

Material Safety Data Sheet (MSDS)

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a) We usually perform macrophage differentiation in T75 flasks and 6-well plates. We haven't tested differentiation in smaller formats. But we assume it will be problematic to thoroughly wash the surface of the wells to remove non-adherent cells after the attachment phase. b) Detachment of the mature macrophages is possible but re-attachment can lead to significant cell loss (30-50%). Please also keep in mind that working in 96/384 well-format has some inherent drawbacks (e.g.; evaporation of media; dry wells; etc.).

This method produces large numbers of adherent monocytes in only 1.5 h. If the washing steps are properly performed 80-90% purity can be expected. The attached cells are “untouched” since no binding of magnetic microbeads has occured. This also excludes phagocytosis of the microbeads by the monocytes an event which is unfavorable with regard to cellular health. In addition the adherence method is time-saving and cost-effective.

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.

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 M1-/M2 Macrophage Generation Media XF we recommend the Nunc plasticware with Nunclon surface - as not only will the detachment efficiency vary (up to 20%); but also the efficiency of the differentiation process itself may be altered.

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.

The Monocyte Attachment Medium allows for the efficient adherence selection (within 1 hr) of monocytes from freshly isolated mononuclear cells while maintaining optimal cell health. The Time-consuming and costly immunomagnetic purification of monocytes prior to DC generation is not necessary and can be skipped when using this medium.

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.

This is not advised. Please seed the freshly isolated CD14-monocytes immediately in the Monocyte Attachment Medium. Adding a culturing step will change the biological characteristics of monocytes very rapidly.

We strongly advise against overnight incubation. The Monocyte Attachment Medium does not contain cytokines/survival factors for the monocytes. If the cells remain in the Monocyte Attachment Medium for longer than 2 hours; they will go into apoptosis and die. The lymphocytes will survive longer. Some of them will attach after such a long time and then cannot be washed off. Therefore; overnight attachment is absolutely inappropriate. However; if time is short; monocyte attachment can be shortened to 1 hour. To do this; it is best to add Monocyte Attachment Medium to the culture vessels in advance and equilibrate it in the incubator. Remove the vessels from the incubator only briefly to add the appropriate amount of concentrated PBMC suspension. Then; even after 1 hour; most of the monocytes are attached.