Technical Library

Recent Entries in Technical Library

• Why is the serum content of PromoCell’s specialized media so low?

  FCS is a natural product consisting of many different components like salts, hormones, vitamins, trace elements, proteins, and enzymes. There can be large lot-to-lot variations between different serum batches regarding the concentrations of growth-promoting factors. The higher the serum content in a culture medium, the higher the impact of the variations on the cell culture system. For this reason, PromoCell has developed several serum-reduced and serum-free media where part of the serum has been replaced by more defined factors like cytokines, hormones or vitamins.Media with reduced serum concentrations have the benefit that they produce more standardized culture conditions over a long time span.

• What culture conditions are required for culturing PromoCell Normal Human Cells in the respective PromoCell media?

  PromoCell Normal Human Cells should be cultured in the appropriate medium at 37°C and 5% CO₂ in a humidified atmosphere.
  Please note: If using cell culture flasks w/o filter cap, unscrew the cap by half a turn to allow sufficient ventilation.

• Can I starve PromoCell HUVECs?

  We have many customers who perform starvation with our HUVECs. Most of them use Endothelial Cell Basal Medium supplemented with FCS (0.5-1% for shorter periods; 5-20% for 24-48 hrs). The cells have to be in a good condition and the experiment should be terminated soon after starvation. Prolonged periods will induce apoptosis.

  Related Links and Documents

  • HUVEC
• Is it possible to perform experimental starvation with PromoCell Normal Human Cells and the PromoCell media?

  Yes, but the experimental starvation conditions have to be determined individually for each cell type. Usually, the cells are maintained in basal medium with reduced growth factor concentrations or lower FBS content. The cells need to be in a good condition and the starvation should be kept as short as possible as prolonged serum and/or growth factor deprivation induces apoptosis.

• What is the exact localization of PromoCell’s HAoAF (Human Aortic Adventitial Fibroblasts)?

  Our HAoAF (C-12380) are isolated from the Adventitia, the outer layer of the aorta. The cells have been characterized as fibroblasts by the expression of fibroblast-specific CD90.

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  • HAoAF
• When subculturing PromoCell’s proliferating HUVEC (T25 flask; C-12250), which TC flask should I use?

  A subconfluent T25-flask typically contains between 0.9 and 1.2 million cells corresponding to 36,000-48,000 cells per cm². It is recommended to count the existing cell number after trypsinization and to calculate the needed number of new flasks.
  Recommended seeding density for HUVEC is 5,000-10,000 cells/cm². This usually corresponds to a split ratio of 1:4-1:6. 1:6 means that you can increase the culture surface by factor 6 (e.g. from 1x T25 to 6x T25 or 2x T75).

  Related Links and Documents

  • HUVEC
• How can I prevent fibroblast contamination in epithelial cell preparations?

  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.

• Why is the growth rate of PromoCell’s HDMEC significantly reduced when I remove hydrocortisone from the Endothelial Cell Growth Medium MV Kit (C-22120)?

  Epidermal Growth Factor (EGF) and hydrocortisone have been reported to have a synergistic effect on the growth of microvascular endothelial cells. Accordingly, if you remove the hydrocortisone, the proliferation of HDMEC is clearly affected.

  Related Links and Documents

  • Endothelial Cell Growth Medium MV
• Where does PromoCell source the heart tissue for HCM isolation?

  The tissue comes from patients who underwent heart transplantation. We obtain a part of the explanted (not the transplanted) heart to prepare cardiac myocytes.

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  • Human Cardiac Myocytes
• For how long can the cardiac myocytes (C-12810) be grown in culture?

  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.

  Related Links and Documents

  • Human Cardiac Myocytes
  • Myocyte Growth Medium