Technical library

No, we do not recommend using PromoCell Media to prepare your own freezing medium.
Our Basal Media are highly complex formulations. Freezing can cause certain components to precipitate irreversibly, which may negatively impact cell viability and post-thaw recovery. Since PromoCell has not evaluated or validated its Media for use as a freezing medium base, we cannot provide recommendations regarding their suitability for this purpose.
For cryopreservation, we recommend using either:

• A standard freezing medium consisting of 70% DMEM, 20% fetal calf serum (FBS), and 10% DMSO, or
• Cryo-SFM Plus (C-29920), PromoCell's serum-free, animal component-free cryopreservation medium.

When working with low-serum or serum-free cell culture systems, the use of a serum-free freezing medium such as Cryo-SFM Plus is particularly recommended to maintain consistency throughout the cell culture workflow and to avoid exposing cells to high serum concentrations that are not present under normal culture conditions.

Yes, PromoCell Normal Human Cells can be re-frozen after thawing and expansion. However, re-freezing is generally not recommended because each additional freeze-thaw cycle can reduce cell recovery, viability, and proliferation potential.
Please note that after an additional freezing cycle, the number of population doublings (PDs) specified in the Certificate of Analysis (CoA) can no longer be guaranteed.
If re-freezing is required, we recommend freezing the cells at a low passage number and using PromoCell Cryo-SFM Plus (C-29920). Cryo-SFM Plus is a defined, animal component-free, and protein-free cryopreservation medium developed to support optimal storage and recovery of primary human cells.
For best results, avoid unnecessary freeze-thaw cycles and establish working stocks from early-passage cultures whenever possible.

No. At day 3 of the dendritic cell (DC) generation protocol, there should be no significant phenotypic difference between the non-adherent cells and the loosely attached dendritic cells.
If you would like to assess the phenotype at this stage, gently rinse the culture surface to detach a portion of the loosely attached cells and combine them with a representative fraction of the non-adherent cells for downstream analysis. This approach provides a sample that reflects the overall dendritic cell population present in the culture at day 3.
Note: Loosely attached cells at this stage are typically considered part of the developing dendritic cell population and can be analyzed together with the non-adherent fraction.
Further reading: For comprehensive guidance on monocyte-derived dendritic cell (moDC) generation, maturation, morphology, and phenotypic characterization, please see our Application Note Generation of Monocyte-Derived Dendritic Cells (moDCs).

For PBMC isolation using 3 ml of Lymphocyte Separation Medium 1077 (C-44010) in a 15 ml conical tube, we recommend loading 3-8 ml of diluted whole blood onto the separation medium.
Before layering, the whole blood should be diluted with PBS at a ratio of 1:2 or 1:3.
 
Example (1:2 dilution):

• Mix 1 volume of whole blood with 1 volume of PBS.
• To obtain 3-8 ml of diluted blood, use 1.5-4 ml of whole blood and add an equal volume of PBS.

As a general guideline:

• 15 ml conical tube: 3 ml Lymphocyte Separation Medium 1077 + 3-8 ml diluted whole blood.
• 50 ml conical tube: 15 ml Lymphocyte Separation Medium 1077 + 15-30 ml diluted whole blood.

For optimal separation, carefully layer the diluted blood onto the Lymphocyte Separation Medium without disturbing the interface.

PromoCell Human Pulmonary Artery Endothelial Cells (HPAEC, C-12241) are isolated from the pulmonary arteries of adult donors. The source tissue is obtained through forensic medicine. Therefore, HPAEC are considered adult primary endothelial cells and are not of fetal or neonatal origin.
Adult HPAECs provide a physiologically relevant model for studying pulmonary vascular biology, including endothelial barrier function, inflammation, and vascular remodeling. They are mainly used in research to study Pulmonary Arterial Hypertension (PAH), Pulmonary Fibrosis, Acute Lung Injury (ALI), and Acute Respiratory Distress Syndrome (ARDS).

Yes. PromoCell Melanocyte Growth Medium M3 (C-24310) can be used for the isolation of primary human melanocytes.
At PromoCell, Melanocyte Growth Medium M3 is routinely used for the isolation and cultivation of Normal Human Epidermal Melanocytes M3 (NHEM M3) from human epidermis. The medium supports the selective growth of melanocytes and is suitable for establishing primary melanocyte cultures following tissue isolation.
For optimal results, please follow the recommended cultivation protocol and use appropriate aseptic cell culture techniques throughout the procedure.