Technical library

The NCCD (Non‑Cancerous Cell Depletion) reagent is a non‑cytotoxic and essential functional component of the Primary Cancer Culture System (PCCS).
 
Purpose of NCCD
NCCD is required to 

• Selectively deplete non‑malignant cells
• Support the selective maintenance of malignant tumor cells

This selective pressure is a core principle of the PCCS and is necessary to ensure reliable enrichment of cancer cells.
 
Use in suspension cultures
NCCD is not dispensable, even if the tumor cells show a suspension growth pattern.
PromoCell recommends:

• Using NCCD throughout the entire culture period
• Applying NCCD both for adherent and suspension cultures

Omitting NCCD may allow non‑malignant cells to persist, which can compromise the selectivity and performance of the PCCS.

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.

Our chondrocytes are derived from patients (~55-80 years) who underwent surgery for total endoprothesis of the hip or knee joint. In most cases this is necessary due to arthrosis. If the tissue shows macroscopic lesions, it is not used for cell isolation.

The amount of media needed per vial depends on the growth characteristics of the cells, the size of the TC vessels and the split ratios used, the frequency of media changes, the type of experiments you perform, etc. It is therefore difficult to give definite quantities. As a rough guideline, 1-2 bottles (500 ml each) are needed for 1 vial of HUVEC.

PromoCell's Osteoblast Basal Medium is an optimized media formulation developed for human osteoblast culture. The exact composition is proprietary.

Generally, you can split differentiated adipocytes. But these cells lose their ability to proliferate after differentiation and you can't expand the culture any more. Therefore it is recommended to plate the cells into the needed vessels (e.g. multiwell plates) prior to induction of differentiation so that trypsinization isn't necessary.

Cell source
PromoCell Normal Human Follicle Dermal Papilla Cells (HFDPC) are isolated from the hair papilla of normal human scalp hair follicles. Dermal papilla cells in adult hair follicles play a key role in dermal-epidermal interactions, which regulate:

• hair production, and
• hair growth cycle events.

Culture characteristics

• HFDPC are cryopreserved at passage 2. 
• When cultured in PromoCell Follicle Dermal Papilla Cell Growth Medium (C‑26501), they can be propagated for at least 10 population doublings
• Typical population doubling times: approximately 20–36 hours

Seeding density and passaging

• Recommended seeding density (after thawing or trypsinization): 5,000-10,000 cells/cm²
• Using a 1:4 split ratio, 4-5 passages can typically be performed.

If you purchase a proliferating culture of human osteoblasts (C-12760), there will be > 500,000 cells in the TC-flask. Once the culture is subconfluent, you will count between 750,000 - 1.1 million cells/T25 (depending on the cell lot).

Our Osteoblast Growth Medium (C-27001) consists of the Basal Medium supplemented with 10% (v/v) FCS, but with no recombinant growth factors. The medium primarily supports the proliferative capacity of normal human osteoblasts. It does not contain osteogenic factors (like dexamethasone and beta-glycerophosphate) that promote differentiation, as many users test their own chemical compounds (growth factors, hormones), or examine the effects of physical strain or sheer stress on the differentiated functions.

The Osteoblast Growth Medium is well-suited as the basis for these applications and can be supplemented with further growth factors if necessary. To specifically induce mineralization, we supply Osteoblast Mineralization Medium (C-27020).