Technical library

Yes, the Skeletal Muscle Cell Growth Medium can also be used for rat, mouse and rabbit SkMC. We recommend to use the medium right after isolation. Cells that were isolated and cultured in a different medium beforehand may have adapted to the other medium. An abrupt medium change causes stress to the cells resulting in reduced growth rates and lower differentiation capacities.

The following criteria are applied for blood donations: a) Donors with light infection like a cold or cough are excluded for one week b) Donors with infections with a temperature above 37.9°C and/ or antibiotics therapy are excluded for 4 weeks c) Blood pressure: exclusion only if systolic blood pressure is < 100 mm Hg or > 180 mm Hg or if diastolic blood pressure is > 100 mm Hg. No exclusion if blood pressure is drug treated and in acceptable range. d) High cholesterol: no exclusion even though medication is used e) Diabetes: exclusion if diabetes type I or use of insulin f) Steroid use: exclusion for 4 weeks after application g) Cancer: exclusion h) Other chronic diseases: exclusion depends on the type of disease (for example chronic heart disease, autoimmune disease)

Our SkMC are proliferating myoblasts that have retained the capacity to differentiate. Upon withdrawal of serum and growth factors, differentiation is induced and the cells form multinucleated syncytia.

For efficient differentiation of our SkMC into myotubes, we recommend to use cells that have undergone a maximum of 4-5 population doublings, i.e. not more than 1 additional subculturing step after thawing the original vial. For more details about differentiation, please see the instruction manual of our Skeletal Muscle Cell Media.

At PromoCell, we get the umbilical cords from our tissue suppliers with no addition of buffers or media. This method prevents the microorganisms from being washed into the blood vessels. Before we start the cell preparation, the umbilical cord is also cut on both ends with a sterile scalpel to provide sterile intersections in addition to the sterile lumen. This method allows us to isolate sterile endothelial cells from umbilical vein and to plate them in antibiotics-free culture media.

The PromoCell quality control procedure includes the cultivation of fibroblasts for 15 population doublings. This is to ensure that the cells can be grown for a minimum of 15 PDs (6-8 passages depending on the split ratio used) but it does not mean that the cells immediately senesce after that point. We don't determine the maximum number of doublings or passages but most NHDF lots will certainly achieve > 20 passages.

Normal osteoblasts, similar to other non-transformed cell types can be expanded in vitro to a certain extent before they are used for experiments. Nonetheless, HOB are generally used at low passages (up to P4) in most labs.

Short protocol:

• Wash the cells with sterile PBS (w/o Ca++/Mg++) or HepesBSS
• Add undiluted accutase to the culture vessel (2 ml per 25 cm²)
• Incubate at room temperature for 5-15 min or at 37°C for faster detachment
• When the majority of the cells has detached, centrifuge the suspension and resuspend the pellet in fresh medium. In most cases, no additional washes or neutralization steps are required.

The source is porcine pancreas.

Our hPC-PL (C-12980) are isolated from microvessels of the human placenta, from the chorionic villi. The number of populations doublings is not determined for each individual cell lot, but in our experience, they can be grown for at least 15 population doublings.