Is the PromoCell Keratinocyte Growth Medium 2 suitable for growth of primary mouse keratinocytes?

Yes, primary mouse keratinocytes grow well in this media when you reduce the CaCl 2 concentration to 0.025 mM - 0.05 mM CaCl 2 (optimal Ca concentration for primary human keratinocytes is 0.06 mM).

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Our subcutaneous HWP are isolated from subcutaneous fat of different localizations, e.g. abdomen, breast, or upper arm. The visceral preadipocytes are isolated from fat surrounding e.g. the pericardium, or from the omentum or mediastinum. The exact localization is specified in the Certificate of Analysis. If you need HWP from a particular localization, please contact our Technical Customer Support prior to placing your order.

The time needed to detach our primary cells depends on many different factors like the cell type, cell density, lot #, trypsin concentration, the efficiency of the washing step before adding the trypsin and the trypsinization temperature. For most cell types we recommend trypsinization at room temperature and direct observation of detachment under the microscope. This way, you can find out your individual trypsinization time and keep the contact time between cells and trypsin to a minimum. Most cells detach after 2-8 min. Please refer to the instructions in the Manual. For some cell types, trypsinization at 37°C or the use of Accutase or another Detachment Solution is recommended.

1) For fluorescence detection (fluorometer): Black plates with clear bottoms; often clear plates will suffice 2) For luminescence detection (luminometer): White/opaque plates 3) For colorimetric detection (photometer): Clear plates

NHEK.f are isolated from tissue samples (foreskin) of single donors, aged between 1-10 years. NHEK.f pooled are prepared from the foreskins of 3 individual donors. The cells of each donor are expanded in separate TC vessels and the cells are pooled after secondary culture, before cryopreservation. Both NHEK from single donor and NHEK pooled are in P2 after thawing.

After isolation and purification of our primary human melanocytes, the cells are checked during quality control whether they show a typical morphology and whether they express the marker Mel-5. Mel-5 is a 75 kDa glycoprotein usually expressed by normal melanocytes. Our Melanocyte Growth Medium (C-24010) has been developed to promote melanocyte growth in vitro. It does not however, completely block the growth of other cells (such as: NHEK or NHDF).

Therefore, it is important to have a pure melanocyte culture from the very beginning.

In contrast, the Melanocyte Growth Medium M3 (C-24310) is much more selective and represses the growth of contaminating cells much better.

Yes, our Skeletal Muscle Cell Differentiation Medium is completely defined. The SupplementMix (C-39366) consists of recombinant human insulin.

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 serum for the production of its growth media.

After addition of the SupplementMix or SupplementPack to our basal medium, 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, primary mouse keratinocytes grow well in this media when you reduce the CaCl₂ concentration to 0.025 mM - 0.05 mM CaCl₂ (optimal Ca concentration for primary human keratinocytes is 0.06 mM).

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 customers have successfully used TC flasks and dishes from all the leading cell culture plastic suppliers to grow PromoCell's primary human cells. We do not have any knowledge whether the dishes from local TC plastic suppliers work in the same way. We recommend to first test whether these brands provide the same good performance as the plastic of the leading manufacturers.

Samples in RNAlater will NOT freeze at -20°C, they are liquid at 4°C and -20°C. The samples can be stored at 4°C for up to 1 month and are stable at room temperature for up to one week. At -20°C, they can be stored indefinitely.