Technical library - RESOURCES

PromoCell guarantee > 15 PD for their HUVEC. The number of passages that can be performed; depends on the dilution factor used during subculture. If you split the cells 1:4; they can perform about 2 population doublings per passage which means that they can be cultured for at least 6-8 passages.

All HDMEC pre-screened lots that we currently have in stock are isolated from foreskin. If you need pre-screened HDMEC from adult donors; please contact the PromoCell Technical Customer Service.

The standard medium for isolation and propagation of our HUVEC HUAEC HPAEC and HSaVEC is Endothelial Cell Growth Medium (C-22010). It contains ECGS an extract from bovine hypothalamus which has mitogenic effects on endothelial cell proliferation. Scientists who prefer a more defined Growth Medium can use Endothelial Cell Growth Medium 2 (C-22011). In this medium ECGS is replaced by VEGF IGF and additional bFGF and EGF to stimulate endothelial cell growth.

Our HCMEC (Human Cardiac Microvascular Endothelial Cells) are not endocardial cells. They are isolated from the capillaries in the heart muscle. Therefore they are in fact microvascular.

Our juvenile HDMEC (C-12210) are isolated from foreskin of young male donors (1-10 years). In contrast; adult HDMEC (C-12212) are derived from different skin localisations like the cheek; temple; or breast. The donors are > 20 years old and are mostly female. Adult HDMEC are the cells of choice when you need cells from a particular part of the body (other than foreskin); or if it is important for your study to use cells from female and/or adult donors.

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.

PromoCell generally culture their HOB on uncoated tissue culture dishes. It is possible however to grow them on collagen type I- or fibronectin-coated dishes as well. Please note: The type of extracellular matrix used may influence the expression of certain genes (e.g. integrins) and thereby affect cellular metabolism. Therefore we recommend to always use the same type of coating matrix for a whole set of experiments.

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.

All Adult Human Blood and Stem cells are analysed by flow cytometry to express defined markers. MSCs are additionally tested for their capacity to differentiate into the osteogenic; chondrogenic and adipogenic lineages. For more details; please see Certificate of Analysis of the respective cell type ("Phenotypic characterization").

It is not necessary to use coated flasks for (most of) our Normal Human Cells but it can be done. As coating with extracellular matrix proteins can affect cellular metabolism; it is recommended to use the same coating material for a complete set of experiments. Cells that need to be grown on coated dishes:

Mesenchymal Stem Cells (C-12974/C-12971/C-12977) need Fibronectin-coating when grown in PromoCell MSC Growth Medium XF (C-28019) and when differentiated in MSC Neurogenic (C-28015); Adipogenic (C-28016); or Osteogenic (C-28013) Differentiation Media.
Human monocyte-derived macrophages (C-12914/C-12916/C-12915/C-12917) must be seeded into Fibronectin-coated culture vessels in combination with PromoCell's M1- and M2-Generation Media XF (C-28055; C-28056).
For efficient induction of osteoblast mineralization with PromoCell's Osteoblast Mineralization Medium (C-27020); the TC plates should be pre-coated with collagen type I.

The population doubling time or generation time (t_g) is usually calculated during the logarithmic phase of growth. It specifies the time (t) in hours needed by the culture to double its cell number. t_g = t / n n: number of population doublings

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