Technical library

PromoCell sources bronchial tissue for the preparation of Human Bronchial Epithelial Cells (HBEpC) from forensic medicine and thoracic surgery.
 
Donor smoking status
For some HBEpC lots, information on the smoking habits of the donors is available. However, this information is not available for all lots.
 
Recommendation
If donor smoking status is critical for your experimental setup, please contact PromoCell Scientific Support prior to ordering so that lot availability can be checked.

Yes. PromoCell specialized media already contain the optimal amount of L‑glutamine.
In most cases, L‑glutamine is included in the form of a stable glutamine derivative to ensure reliable performance and shelf life.

Important note
Please do not add additional L‑glutamine to PromoCell media. Excess L‑glutamine can be toxic to cells and may negatively affect cell viability and experimental results.

The lead time for customized media at PromoCell is typically 4-8 weeks. 
Our Scientific Support team can help evaluate the feasibility of custom media formulations and advise on available customization options for your specific application.

Pericytes have been shown to possess multilineage differentiation potential. Depending on the experimental context and microenvironment, they can differentiate into a variety of mesenchymal and vascular cell types, including:

• Preadipocytes
• Osteoblasts
• Chondroblasts/chondrocytes
• Fibroblasts/myofibroblasts
• Vascular smooth muscle cells
• Phagocytic cells

This differentiation plasticity highlights the role of pericytes as multipotent progenitor‑like cells involved in tissue remodeling, regeneration, and vascular biology.
 
Further information:
Diaz‑Flores et al.; Histol Histopathol 2009 Jul;24(7):909

Fetal calf serum (FCS/FBS) is a natural product that contains a complex and variable mixture of components, including salts, hormones, vitamins, trace elements, proteins, and enzymes. As a result, there can be substantial lot‑to‑lot variations in the concentration of growth‑promoting factors between different serum batches.
The higher the serum content in a culture medium, the greater the impact of this variability on cell growth, phenotype, and experimental reproducibility.
 
PromoCell approach
To minimize these effects, PromoCell has developed a range of serum‑reduced and serum‑free media, in which parts of the serum are replaced by more defined components, such as cytokines, hormones, and vitamins.
 
Benefit of low‑serum formulations
Media with reduced serum content provide:

• more standardized and reproducible culture conditions
• greater consistency over long‑term experiments
• reduced batch‑to‑batch variability

PromoCell Normal Human Cells should be cultured in the appropriate PromoCell medium at 37°C and 5% CO₂ in a humidified atmosphere.
Please note: When using cell culture flasks without a filter cap, the cap should be unscrewed by approximately half a turn to ensure sufficient gas exchange and ventilation.

Yes. Many customers perform serum starvation experiments with PromoCell HUVECs.
Most commonly, Endothelial Cell Basal Medium supplemented with FBS, but without any additional growth supplements, is used as the starvation medium. Typical conditions include:

• 0.5–1% FBS for short-term serum starvation
• 5–20% FBS for longer incubation periods (e.g. 24-48 hours). Although these conditions are not serum-reduced, cells remain deprived of the growth factors normally present in Endothelial Cell Growth Medium and can therefore be considered growth factor-starved.

Important considerations

• Cells should be in good physiological condition before starting starvation.
• Experiments should be terminated shortly after the starvation period.
• Prolonged starvation can induce apoptosis and compromise cell viability.

Yes. Experimental starvation can be performed with PromoCell Normal Human Cells using PromoCell media. However, starvation conditions must be optimized individually for each cell type.
In practice, starvation is typically achieved by:

• Culturing cells in basal medium
• Reducing growth factor concentrations
• Lowering the FBS content (if serum is used)

Important considerations

• Cells should be in good overall condition before starting starvation.
• Starvation periods should be kept as short as possible, as prolonged deprivation of serum and/or growth factors can:
  • Induce cell stress
  • Trigger apoptosis
  • Compromise experimental outcomes

Careful optimization of duration and conditions is therefore essential.

PromoCell Human Aortic Adventitial Fibroblasts (HAoAF, C‑12380) are isolated from the adventitia, the outermost layer of the human aorta. 
The cells are characterized as fibroblasts by the expression of the fibroblast‑specific marker CD90, confirming their stromal fibroblast identity.

A subconfluent T25 flask of proliferating HUVEC typically contains approximately 0.9–1.2 million cells, corresponding to about 36,000 - 48,000 cells/cm².
 
Recommended approach
PromoCell recommends counting the cells after trypsinization, and calculating the required number and size of new flasks based on the recommended seeding density.
 
Seeding density
Recommended seeding density for HUVEC: 5,000 - 10,000 cells/cm²
 
Typical split ratios
This seeding density usually corresponds to a split ratio of 1:4 to 1:6.
1:6 split ratio means that the total culture surface can be increased by a factor of 6, for example:

• from 1 × T25 to 6 × T25, or
• from 1 × T25 to 2 × T75

The exact choice of flask size depends on your experimental needs and available culture capacity.