Technical library

Secretory granules called Weibel-Palade bodies (WPBs) containing Von Willebrand factor (vWF) are more linked to the formation of a confluent endothelial monolayer. Research shows that vWF expression is dynamic and highly dependent on the cell culture conditions such as confluence and passage number. Therefore, having vWF as a quality control marker for each lot of ECs is not really necessary.

Further Information

Howell et al.; Mol Membr Biol. Nov-Dec 2004;21(6):413-21

Yes, Accutase Solution can be defrosted, aliquoted, and then refrozen. Defrosting: Accutase should be defrosted overnight in the refrigerator or placed in a tub of cold tap water. Do not defrost in a 37°C water bath. Stability: Once thawed, it is stable for at least 2 months in the refrigerator if stored promptly after use.

Yes, it is possible to refreeze them.

This is not advised. Please seed the freshly isolated CD14-monocytes immediately in the Monocyte Attachment Medium. Adding a culturing step will change the biological characteristics of monocytes very rapidly.

The PBS buffering enhances the Alizarin Red staining (precipitation of the dye) and makes it more intense. Leave the PBS on the cells after staining/washing and analyze the sample immediately, as the dye may bleed upon prolonged storage without embedding.

The use of antibiotics creates a false sense of security and allows users to develop poor aseptic techniques. This leads to low-level contamination with partially resistant bacteria occurring but being overlooked for a time. This then leads to cells with undetected contamination being cultured for extended periods of time, increasing the risk that contamination will spread throughout the laboratory and eventually antibiotic-resistant strains of bacteria may develop. Mycoplasma infections can also occur more easily, as they are often introduced along with contaminants such as bacteria and fungi. Last but not least, antibiotics are known to have negative effects on the metabolism of eukaryotic cells - more details on this topic can be found in our blog article "Antibiotics in cell culture: friend or enemy".

Further Information

Blog article: Antibiotics in Cell Culture: Friend or Enemy?

The supplement should be at room temperature when added to the MSC Adipogenic Basal Medium 2. It may also be beneficial to invert the tube a few times to bring precipitates back into solution. Please note: It is not recommended to filter the basal medium, supplements, or complete medium, as components that induce or promote differentiation may be removed, resulting in a low differentiation rate when using the medium.

Yes, there are a few differences:

• We recommend replacing the MSC Growth Medium XF (C-28019) 3-4 h after seeding, as opposed to 16-24 hours after seeding for most other cell types/growth media, including MSC Growth Medium 2.
• IF MSC Growth Medium XF, MSC Neurogenic Differentiation Medium, MSC Adipogenic Differentiation Medium 2, or MSC Osteogenic Differentiation Medium are used, the TC vessels must be pre-coated according to the instruction manual.
• We strongly recommend using Accutase (C-41310) for cell detachment instead of Trypsin. If Trypsin is used, the contact time should not exceed 2 min.

Light flocculation may be seen upon thawing the supplements containing ECGS/heparin or BPE. This does not affect the activity of our media. Optionally, the precipitate can be removed by centrifugation under sterile conditions. We recommend to thaw the supplements (SupplementMix or SupplementPack) at 15-25°C.

Our HPAEC and HPASMC are isolated from the proximal pulmonary artery.