Technical Library
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PromoCell's Endothelial Cell Growth Medium (C-22010) is a complete medium that can be used for the culture of HUVEC after addition of the SupplementMix.
Addition of extra FCS is not necessary. The SupplementMix contains FCS (2% v/v final concentration); recombinant growth factors; hormones; and a bovine brain extract that together have mitogenic effects on endothelial cells.
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).
It actually depends on the culture conditions whether the cells remain in suspension or attach to the surface.
When grown in our serum-free; xeno-free HPC Expansion Medium XF (C-28021); the cells remain in suspension.
You can use our standard DetachKit (C-41200) to subculture the human nasal epithelial cells. Some customers still prefer to use our DetachKit-2 (C-41202) as it has a lower trypsin/EDTA concentration but tests in our cell culture lab haven't revealed any adverse effects when using C-41200.
PromoCell's HDLEC are isolated from skin (dermis). When isolated from juvenile donors (C-12216); the exact localization is foreskin. When derived from adult donors (C-12217); the localization depends on the type of surgery; e.g. breast or temple. You can find the information on the exact localization in the Certificate of Analysis.
Our HDLEC are tested to be positive for CD31; podoplanin; and Prox-1 and are delivered in P2. The recommended culture medium is Endothelial Cell Growth Medium MV2 (C-22022).
No we don't determine the ratio of HDLEC and HDBEC in our HDMEC lots. From our experience the percentage of HDLEC is highly lot-dependent and can vary between 5 and 60%.
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.
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.
The medium supplemented with Cytokine Mix E is stable for 2 weeks if stored protected from light at 2-8°C.
Soluble receptors are post-translationally modified and generally contain disulfide bonds and glycosylation sites. If produced in E. coli the proteins don't show any biological activity.
Our Human Mesenchymal Stem Cells and Human Pericytes are cryopreserved at the end of secondary culture (P1). After thawing they are in P2.
Serum turbidity is usually caused by cryoprecipitation of lipid components during freezing and thawing. The more times the serum is subjected to freeze/thaw cycles the more turbidity is noticed. It can be minimized by freezing the serum in aliquots at -20°C and thawing these aliquots individually at the time of use.