Technical library

No. Hematopoietic progenitor cells (e.g. CD34⁺ or CD133⁺ cells) will not proliferate in Hematopoietic Progenitor Cell Expansion Medium XF (C‑28021) without additional cytokines.
 
Required supplementation
For efficient expansion, the medium must be supplemented with either:

• Cytokine Mix E (C‑39890), or
• a user‑defined cytokine mixture with equivalent functionality.

Cytokine Mix E is a ready‑to‑use formulation containing recombinant human Thrombopoietin (TPO), Stem Cell Factor (SCF), Flt3‑Ligand, and Interleukin‑3 (IL‑3).
 
Expansion performance
When used in combination with Cytokine Mix E, Hematopoietic Progenitor Cell Expansion Medium XF supports strong progenitor cell expansion for at least two weeks.

All PromoCell Adult Human Stem and Blood Cells are phenotypically characterized by flow cytometry to confirm the expression of defined, cell‑type‑specific surface markers.
 
Additional characterization of MSCs
In addition to marker analysis, Mesenchymal Stem Cells (MSCs) are functionally tested for their ability to differentiate into the three mesenchymal lineages:

• osteogenic
• chondrogenic
• adipogenic

Where to find the details
The exact markers tested for each cell type are listed in the Certificate of Analysis (CoA) of the respective product.

The cells will die. Hematopoietic Progenitor Cell Expansion Medium XF always must be supplemented with Cytokine Mix E or an appropriate cocktail of cytokines.

The difference between PromoCell DC Generation Medium XF (C‑28052) and DC Base Medium XF (C‑28054) is the presence of cytokines and the level of preparation:

• DC Base Medium XF (C‑28054) consists of 250 ml basal medium plus SupplementMix, but does not contain cytokines. It must therefore be supplemented with cytokines according to the user’s individual experimental requirements.
• DC Generation Medium XF (C‑28052) is ready‑to‑use and consists of 250 ml basal medium, SupplementMix, and the appropriate cytokines required for dendritic cell generation. No additional supplementation is needed before use.

Both DC Base Medium XF and DC Generation Medium XF feature a xeno‑free formulation.

Our DC Generation Medium XF (C-28052) has a xeno-free formulation. It provides a complete media system (ready-to-use, all cytokines included) and shows efficient and reproducible in vitro maturation of moDCs from freshly isolated peripheral blood monocytes.

Yes. PromoCell Lymphocyte Separation Medium 1077 (C‑44010) is endotoxin‑tested.
The product specification is < 0.3 EU/ml.

It is possible to culture the airway organoids in 96-well U-Bottom plates for suspension cells (e.g., Greiner Bio-One # 650185). A detailed protocol for the use of 96-well plates can be found in our AppNote. We have not tested 384-well plates or other commercially available plates.

ROCK inhibitors (ROCKi), such as Y‑27632, are widely used to enhance epithelial cell and keratinocyte progenitor proliferation and to improve cell survival through a process known as conditional reprogramming. Importantly, this effect is reversible.
In addition, ROCKi is known to:

• Increase seeding efficiency on plastic surfaces
• Support early survival of epithelial cells during stressful steps (e.g. dissociation, seeding)

ROCKi in airway epithelial cells and organoids
The effects of ROCKi‑mediated conditional reprogramming have also been described for airway epithelial cells (EpCs) and airway organoid cultures, as reported in the literature:

• https://pubmed.ncbi.nlm.nih.gov/31871466/
• https://pubmed.ncbi.nlm.nih.gov/35169685/

PromoCell recommendation
For human airway organoid cultures, the use of Y‑27632 ROCKi is optional, as organoids are able to form even without ROCKi. However, 3‑D culture systems can be sensitive to factors such as:

• the choice of plastic
• the extracellular matrix (ECM) used

Under these conditions, the addition of ROCKi may help to increase robustness and reproducibility of the system, particularly during the initial phases of organoid establishment.

Possibly. The size of the organoids may depend on the access to nutrients in the gel. For reference, https://pubmed.ncbi.nlm.nih.gov/31173716/ For best results we recommend the following:

• Avoid nutrient gradients. We recommend using a small, drop-like BME gel bead and high volumes of medium. Change the medium every day as we’ve observed that less frequent medium changes results in yellow culture medium indicative of a pH shift likely from an increase in cellular metabolism.
• For optimal cell distribution, it is important to control the “gelling” or setting of the ECM gel. Therefore, the temperature is critical! BME is liquid in the cold (4°C) and solidifies at 37°C. The gelling must be done in a very short time, otherwise the cells in the BME would sink to the bottom.

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.