Technical library

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.

Epidermal Growth Factor (EGF) and hydrocortisone are known to have a synergistic effect on the proliferation of human dermal microvascular endothelial cells (HDMEC).
When hydrocortisone is removed from the Endothelial Cell Growth Medium MV Kit (C‑22120), this synergistic interaction is lost. As a consequence, cell proliferation is clearly reduced, even though EGF is still present.
 
Conclusion
For optimal growth and proliferation of HDMEC, hydrocortisone should be included as specified in the complete Endothelial Cell Growth Medium MV formulation.

PromoCell Fibroblast Growth Medium (C‑23010) is a very low‑serum medium (containing 0.5% FBS in the basal medium) and is therefore not subject to the pronounced lot‑to‑lot variability commonly observed with DMEM supplemented with 10% FBS.
As a result, using C‑23010 allows for more standardized and reproducible culture conditions when growing juvenile fibroblasts (C‑12300), while still supporting robust cell growth.

After adding the Skeletal Muscle Cell Differentiation Medium, myoblasts will start to differentiate into myotubes and stop growing. The cells should not be split anymore. It is recommended rather to plate the SkMC into the needed vessels, (e.g. multiwell plates), prior to induction of differentiation and to perform studies directly on differentiated cells. If it is necessary for your tests to detach the myotubes and they are difficult to trypsinze, you can use a "rubber policeman".

Fibroblast contamination cannot be completely avoided in primary cell cultures. As epithelial cells attach more firmly than fibroblasts, it is possible to perform partial trypsinization to remove the fibroblasts. This is done by adding trypsin/EDTA to the TC dish for 2-4 min. When the fibroblasts detach, the enzyme is inactivated and the suspension with the fibroblasts aspirated. The remaining epithelial cells are washed twice with buffer and their culture is continued in the respective Growth Medium.

In most cases, PromoCell Human Cardiac Myocytes (HCM) are isolated from heart tissue obtained during heart transplantation procedures.

Specifically:

• The tissue is taken from the explanted heart (the removed native heart), not from the transplanted donor heart.
• In some cases, heart tissue is also obtained from LVAD (Left Ventricular Assist Device) surgeries, where cardiac tissue becomes available during the implantation procedure.
• All tissue sourcing follows ethical guidelines and applicable regulations.

PromoCell guarantees 15 population doublings (PDs) if the HCM are grown in Myocyte Growth Medium. Depending on the cell lot and the culture conditions, the cells can be maintained in culture for > 6-8 passages corresponding to a period of 1-2 months.

The recommended culture or differentiation medium depends on the specific stem or blood cell type. For each product, the appropriate media are specified in the Instruction Manual provided for the cells.
Please always follow these cell‑type‑specific recommendations, as the use of non‑recommended media may affect cell growth, differentiation efficiency, and functionality.

Growth medium requirements vary depending on the cell type. Please consult the corresponding Product Manual and refer to the "Specifications" section for the recommended Growth Medium. Using the recommended medium is important for maintaining optimal cell viability, growth, and functionality.

The standard PromoCell protocol recommends performing trypsinization at room temperature (RT) while monitoring cell detachment under the microscope.
Prolonged trypsinization at 37°C may lead to irreversible cell damage and reduced cell viability.
 
Exceptions
For certain cell types, trypsinization at 37°C may be recommended.
In these cases, the Instruction Manual of the respective cell type specifies the temperature and exact duration of trypsinization.