Technical library

Yes. The orientation can be triggered by the use or lack thereof of ECM. Without the use of an ECM, the organoids will have a higher outward oriented ratio. You can find a protocol here https://pubmed.ncbi.nlm.nih.gov/30811997/, where cells were first embedded in ECM gel and afterwards ECM was dissociated, and free organoids were re-seeded in suspension without an ECM. * Please note that we have not tested this method in our labs and thus cannot guarantee it.

PromoCell is using a pan-cytokeratin antibody rather than a specific type of cytokeratin for the quality control of our keratinocytes.

We guarantee > 500,000 viable cells per vial. To do this, we need to freeze more than 500,000 cells, as we do not yet know the viability after thawing at the time of freezing. For technical/organizational reasons, the initial cell number may also vary from lot to lot, so 2 lots with the same viability may not necessarily contain the same number of cells.

Yes, there is a reference (Campuzano et al.; J Immunol. 2020 Jun 15;204(12):3296) where bone marrow-derived macrophages from mouse were detached using our Macrophage Detachment Solution (40 minutes at 4°C).

Further Information

Campuzano et al.; J Immunol . 2020 Jun 15;204(12):3296

PromoCell's Normal Human Epidermal Keratinocytes (NHEK) from juvenile donors are isolated from both the epidermis of the outer and mucosal (i.e. inner) layers of the foreskin. Thus, each vial contains a mixture of mucosal and cutaneous keratinocytes.

Even non-activated macrophages do release a certain amount of cytokines. Furthermore, you would have to be sure that the release of a certain cytokine is a direct consequence of the activation. Therefore, we do not think it is possible to have a general negative control for the cytokine release.

No, we do not provide data about the cytokine profile of our M1/M2 macrophages after activation and we do not provide any further data beyond the scope of our quality control.

You may find all information regarding the activation and the cytokine concentrations in table 1 (page 5) of our Application Note.

We did not plate the cryo-macrophages in 96-well plates. However, we heard from other customers that they have successfully used our macrophages in this kind of plate.

Scientific findings from different groups, as well as our own results indicate that the presence or absence of α-SMA is not a valid indicator for the composition of a given cell population. Research findings and our inhouse data suggest that endothelial cells are not α-SMA-negative under all circumstances. α-SMA negativity is not an intrinsic property of endothelial cells but can vary depending on extrinsic influences. Lot-specific values for α-SMA in our Certificates of Analysis (CoAs) are therefore not considered meaningful.

Further Information

Ando et al.; Am J Physio l. 1999 May;276(5):H1755-68

Lu et al.; Stem Cells Dev . 2004 Oct;13(5):521-7

Azuma et al.; Biochem Biophys Res Commun . 2009 Mar 13;380(3):620-6

Frid et al.; Circ Res . 2002 Jun 14;90(11):1189-96

Kovacic et al.; J Am Coll Cardiol . 2019 Jan 22;73(2):190-209

Jones et al.; Am J Respir Cell Mol Biol . 1999 Apr;20(4):582-94

Ma et al.; Front Cell Dev Biol . 2020 Apr 21;8:260

Kocher and Madri; In Vitro Cell Dev Biol . 1989 May;25(5):424-34

Amberger et al.; FEBS Lett . 1991 Aug 5;287(1-2):223-5