Technical library

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.

Precoating of culture vessels with ECM proteins does not have adverse effects on the cells but has been reported to influence the cellular expression pattern. It is therefore recommended to use the same culture conditions e.g. fibronectin- collagen- or gelatin-coating for a whole set of experiments to be able to compare the results.

No it is not necessary to use coated flasks therefore we don´t recommend their usage in the Instruction Manual. However for special applications some of our customers use collagen-coated dishes.

We source the bronchial tissue from forensic medicine and from thoracic surgery. For some lots; the smoking habits of the donors are known. Please contact our Technical Customer Service before ordering the cells if you need this information.

Our Nasal Epithelial Cells are isolated from nasal septum or adenoids.

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

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 amount of media needed per vial depends on the growth characteristics of the cells the size of the TC vessels and the split ratios used the frequency of media changes the type of experiments you perform etc. It is therefore difficult to give definite quantities. As a rough guideline 1-2 bottles (500 ml each) are needed for 1 vial of HUVEC.

During our quality control we do not determine the presence or maintenance of respective stem cell markers in the tumorspheres. We use a functional approach instead; by culturing the tumorspheres over serial passages in C-28070. Tumorsphere formation requires the biological features of Anoikis resistance and self-renewal and therefore indicates the presence of cancer stem cells (CSC)/cancer initiating cells (CIC) in the spheres. 3D tumorsphere culture thus allows scientists to study the biology of CSCs without any background knowledge on CSC markers.

Please see our attached alternative product guides for media and cells. Please note: Our media do not contain antibiotics. For optimal cell growth; we recommend to refrain from using antibiotics. However; when a sterile environment cannot be 100% ensured; it may be advisable to add antibiotics to your media so as to protect the cultures from potential microbial infections.

There are several factors that can influence successful transfections; e.g. viability and density of the cells; choice of the transfection reagent; quality and type of the transfected molecules (plasmids; siRNA; oligonucleotides); as well as the culture medium and supplements used. When using PromoCell Endothelial Cell Growth Media for cell transfection; please follow the instructions below: Heparin; which is included in our Endothelial Cell Growth Medium (C-22010/C-22110); Medium 2 (C-22011/C-22111) and Medium MV (C-22020/C-22120) may reduce the transfection efficiency.  We therefore recommend to use our heparin-free Endothelial Cell Growth Medium MV2 (C-22022/C-22121). Alternatively; you may use Endothelial Cell Growth Medium Kit (C-22110); Medium 2 Kit (C-22111); or Medium MV Kit (C-22120) without adding the ECGS/heparin supplement to the Basal Medium. Please note: Before and after transfection; the cells should be cultured in complete Growth Medium including heparin to ensure optimal growth.

Yes; we have received a customer feedback that our MSC Growth Medium 2 also works for rat MSCs. The rat cells grow nicely in this medium and have a good viability.