Should I store the cryopreserved cells in the liquid phase or gas phase of liquid nitrogen?

In principle, both types of liquid nitrogen storage are acceptable, each having its advantages and disadvantages. Liquid phase storage provides a consistent temperature of -196°C, a longer holding time and a greater vial capacity but involves the risk of contamination issues. Storage in the gas phase is very safe with respect to contaminations but the holding time of the cells is shorter and the vial capacity is reduced.

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At manufacture, ECGS is adjusted to a protein content of 3 mg/ml. For Human Endothelial Cells and Microvascular Endothelial Cells the optimal concentration of ECGS is 2 ml/500 ml medium, corresponding to 6 mg extracted protein/500 ml medium. ECGS/H is additionally supplemented with 22.5 mg/ml heparin, corresponding to a final concentration of 45 mg heparin/500 ml medium.

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.

The three components of our DetachKit (HepesBSS, Trypsin/EDTA, TNS) are stable for 1 year from the date of manufacture when stored at -20°C.

Once thawed for usage, they should be stored at 4-8°C and can be used for 6 weeks. Please avoid repeated freeze/thaw cycles.

Our Normal Human Cells have been cultured and tested in our growth media and have adapted to these conditions. Using other media may yield unsatisfactory results due to suboptimal supplies of nutrients and growth factors. PromoCell can only guarantee good cell growth (as stated in the Certificate of Analysis), when the cells are grown in the recommended media.

PromoCell Growth Media have special formulations and are much more complex than DMEM or RPMI + FBS. In comparison to immortalized cell lines, primary cells have much higher nutrient and growth factor requirements. Classical media do not usually achieve good performances with our cells.

PromoCell cell pellets are made from > 1 million cells and are dissolved in 200 µl RNAlater.

Our Melanocyte Growth Medium M3 allows the serum-, BPE- and PMA-free cultivation of Normal Human Epidermal Melanocytes (NHEM) without additional coating of the TC plastic. To maintain the cells in a robust adherent pro-proliferative phase, we highly recommend the passaging of cells at 70-90 % confluency. It is known from the literature that high cell densities of NHEM can promote the growth of 3D spheroids. Therefore, too high confluencies should be avoided.

We strongly recommend using gentamicin at a concentration of 50 μg/ml. Many other antibiotics (such as penicillin or streptomycin) inhibit the growth of primary cells (including primary cancer cells), especially at the clonal level or at low cell density. Therefore, you should be very cautious with antibiotics and antifungals, and we recommend sticking with gentamicin. If an antifungal agent is required, we recommend 10 μg/ml caspofungin (100x stock solution = 1 mg/ml in water).

Tumor-associated macrophages (TAM) represent an important constituent of the tumor microenvironment. After being educated by cancer cells, TAM adopt an anti-inflammatory, pro-tumor and pro-metastatic M2-like phenotype.

Our Primary Cancer Cell Medium D-ACF allows for the isolation and culture of tumor associated macrophages (TAM) from solid tumors while reliably preventing stromal overgrowth.

The isolated TAM are non-proliferating adherent cells and can be cultured for at least two weeks.

The NCCD (Non-Cancerous Cell Depletion) reagent is a non-cytotoxic and functional component of the PCCS - necessary for the selective maintenance of malignant cells.

It should be used throughout the whole culture in our PCCS, even for suspension cultures.

No, the supplied serum is not heat-inactivated.

In principle, both types of liquid nitrogen storage are acceptable, each having its advantages and disadvantages.

Liquid phase storage provides a consistent temperature of -196°C, a longer holding time and a greater vial capacity but involves the risk of contamination issues.
Storage in the gas phase is very safe with respect to contaminations but the holding time of the cells is shorter and the vial capacity is reduced.