Technical library

Yes. PromoCell cell pellets (C‑14***) are an easily accessible source of DNA, RNA, and proteins.
Please note that cell pellets are not viable and cannot be used for cell culture.

No. PromoCell cell pellets (C‑14***) cannot be revived and cannot be used to establish a growing culture. The cell pellets are supplied frozen at -20°C and are intended for RNA and protein isolation and subsequent downstream analyses.
For applications requiring viable, expandable cells, cryopreserved cells from the same donors are available. If you require matched viable cells, please contact PromoCell Scientific Support for assistance.
 

Slow growth after subculture is often caused by over‑trypsinization, which can damage the cells and reduce their viability.
In addition, suboptimal culture or handling conditions may also affect growth, for example:

• Insufficient inactivation of trypsin
• EDTA not removed by centrifugation after detachment
• Incorrect seeding density
• Use of non‑recommended media or supplements

To ensure optimal growth, please follow the recommended subculture protocol and culture conditions provided in the Product Manual.

Poor or absent attachment after thawing is usually caused by stress during freezing, storage, or thawing, or by suboptimal culture or handling conditions immediately after seeding.
Common reasons include:

• Inappropriate freezing or storage conditions (e.g. incorrect freezing protocol or storage outside liquid nitrogen)
• Suboptimal thawing procedure (e.g. slow thawing, prolonged exposure to DMSO, incomplete DMSO dilution after thawing)
• Inadequate culture conditions such as incorrect or incomplete growth medium, expired or improperly stored media or supplements, incorrect incubator settings (temperature, CO₂, humidity), unsuitable cell culture plasticware or missing coating (if required).

The difference between PromoCell DC Generation Medium Ready‑to‑use (C‑28050) and DC Base Medium (C‑28053) lies in the presence of cytokines and the degree of preparation:

• DC Base Medium (C‑28053) 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 needs.
• DC Generation Medium Ready‑to‑use (C‑28050) is a fully prepared medium and consists of 250 ml basal medium, SupplementMix, and the required cytokines. No additional supplementation is required before use.

PromoCell Monocyte Attachment Medium (C‑28051) is used when working with freshly isolated peripheral blood mononuclear cells (MNC) and a rapid, efficient adherence‑based selection of monocytes is required.
 
Purpose
The Monocyte Attachment Medium enables:

• Efficient adherence of monocytes within ~1 hour
• Selective enrichment of the monocyte fraction
• Preservation of optimal cell health

Key advantage
When using Monocyte Attachment Medium, time‑consuming and costly immunomagnetic monocyte purification steps are not required. The medium allows direct progression to downstream applications, such as macrophage or dendritic cell generation, without prior monocyte isolation.
 
Further information:
Detailed protocols for macrophage and dendritic cell generation from freshly isolated monocytes are provided in the corresponding PromoCell Application Notes.

• Generation of monocyte derived Dendritic Cells (moDCs)
• Differentiation of M1- or M2-Macrophages from PBMC/Monocytes

To differentiate skeletal muscle cells (SkMC) into myotubes, PromoCell recommends using early-stage cells with no more than 4-5 population doublings (PDs) after thawing.
 
Recommended differentiation workflow
1. Cell stage and expansion
- Use SkMC ideally at passage P2 or P3
- Cells should not exceed 4-5 population doublings
2. Growth phase
- Culture cells in Skeletal Muscle Cell Growth Medium (C‑23060)
- Grow cells to 60-80% confluency
3. Induction of differentiation
- Switch to serum‑free Skeletal Muscle Cell Differentiation Medium (C‑23061)
- This medium change induces differentiation and the formation of multinucleated syncytia (myotubes)
4. Maturation phase
- After 5 days, switch back to Skeletal Muscle Cell Growth Medium (C‑23060)
- Continue culture for an additional 8 days to complete differentiation

Best differentiation results for PromoCell skeletal muscle cells (SkMCs) are obtained when cells have reached 60-80% confluency.
Recommended differentiation workflow:

• At 60-80% confluency, aspirate the Growth Medium.
• Replace it with Skeletal Muscle Differentiation Medium to initiate differentiation.
• After 2–8 days, formation of multinucleated syncytia (myotubes) can typically be observed.
• For stable and robust differentiation, switch back to Skeletal Muscle Cell Growth Medium after 5 days of incubation in Differentiation Medium.

Yes. PromoCell Skeletal Muscle Cell Growth Medium (C‑23060) can also be used to culture primary mouse, rabbit, canine, and sheep skeletal muscle cells (SkMCs).
For best results, we recommend using the medium immediately after cell isolation. Cells that have been previously cultured in a different medium may have adapted to those conditions. An abrupt medium change can stress the cells and may lead to reduced growth rates and impaired differentiation capacity.

Peripheral blood used by PromoCell for the isolation of mononuclear cells from peripheral blood (MNC‑PB) and monocytes is obtained from carefully screened donors. A defined set of selection and exclusion criteria is applied prior to blood donation.
 
Donor exclusion criteria
The following conditions lead to temporary or permanent exclusion from blood donation:
a) Acute infections
- Mild infections (e.g. cold or cough): exclusion for 1 week
- Infections with body temperature > 37.9°C and/or antibiotic treatment: exclusion for 4 weeks
b) Blood pressure
- Exclusion if:
  systolic blood pressure < 100 mm Hg or > 180 mm Hg, or
  diastolic blood pressure > 100 mm Hg
- No exclusion if blood pressure is medically treated and within an acceptable range
c) Metabolic conditions
- High cholesterol: no exclusion, even if medication is used
- Diabetes: exclusion in cases of type I diabetes or insulin therapy
d) Medication use
- Steroid treatment: exclusion for 4 weeks after application
e) Severe or chronic diseases
- Cancer: exclusion
- Other chronic diseases: exclusion depends on the specific condition (e.g. chronic heart disease, autoimmune diseases)