Technical Library

For the M2 Macrophage Generation Medium; it is extremely important that the shelf life of 2 weeks (after addition of the cytokines) is not exceeded; the yield will quickly decrease thereafter. It is best to use the M2 medium as fresh as possible to avoid discrepancies between M1 and M2 yield.

Yes; our Lymphocyte Separation Medium 1077 is endotoxin-tested. The specification for the product is < 10EU/ml.

Yes we have received a customer feedback that our 3D Tumorsphere Medium XF (C-28070) has been successfully used for tumorsphere formation of mouse cell lines.

MSC Growth Medium 2 (C-28009) is an optimized medium formulation with reduced serum content to allow for more standardized culture conditions (considerably lower lot to lot variation). You can replace MSC Growth Medium by MSC Growth Medium 2. Coating of culture vessels is not necessary. We recommend to plate the cells (hMSCs from bone marrow; adipose tissue; or umbilical cord) at 4;000 cells/cm².

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.

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.

The Macrophage Detachment Solution (C-41330) directly affects the cell membrane. HSA in the Wash Buffer supports regeneration of the cell membrane and protects the cells during the critical phase directly after detachment from detrimental effects.

At PromoCell we guarantee for our primary human cells ≥ 500;000 viable cells after thawing. For this; we dispense > 500;000 cells per cryovial before cryopreservation as there will always be a certain percentage of dead cells after freeze/thaw. In order to know the number of cells that survived the procedure; we defrost a representative number of vials per lot during QC; determine the cell viability using an electronic counting device and then calculate the number of viable cells that can be recovered after thawing. Both numbers - the calculated number of viable cells and the viability - can be found on the lot-specific Certificate of Analysis (CoA) that can be downloaded from our website. Example: When the CoA indicates 600;000 viable cells and a viability of 80%; this means that the vial actually contains 750;000 cells (viable + dead); 80% thereof (600;000) were viable after thawing in our QC. We do not indicate the total number of cells per vial but just the number of expected viable cells which can be recovered when the recommended thawing protocol is used. You don't have to calculate any viabilities by yourself. When the recommended plating density for your cell type is 5;000 - 10;000 cells/cm²; then the 600;000 viable cells can be plated e.g. in a T75 (corresponding to 8;000 cells/cm²) or in a T75 + a T25 (corresponding to 6;000 cells/cm²).