In principle all microvascular endothelial cells should be able to migrate proliferate and form tubes or sprouts in an appropriate assay after angiogenic stimulation. As this is not part of our routine quality control procedure we cannot tell for sure whether all cell lots will respond to angiogenic stimuli. However PromoCell also supplies HDMEC pre-screened (C-12215) that are especially tested for a positive VEGF response.

These cells are frozen at the end of 2^nd culture. Thawing and seeding results in passage 2 (3^rd culture). We recommend that they be used for differentiation experiments not later than passage 5. The differentiation potential of hMSC in vitro is reduced with ongoing population doublings; meaning the earlier differentiation is induced; the higher the differentiation rates.

PromoCell provides two types of Renal Epithelial Cells: Human Renal Epithelial Cells (HREpC) and Human Renal Cortical Epithelial Cells (HRCEpC). HREpC are isolated from the adult kidney and stain positive for cytokeratin. They comprise a heterogeneous population of renal epithelial cells. HRCEpC are isolated from the cortex of the kidney and comprise cells from proximal and distal tubuli. They also stain positive for cytokeratin.

Short protocol:

Trypsinize the cells as usual
Centrifuge and resuspend in suitable cold freezing medium at a density of 1-4 x 10⁶ cells/ml
Cool down the cells slowly to -80°C (approx. -1°C per min). We recommend to use "Mr. Frosty" from Nalge or "CoolCell" from Biocision; which both provide gradual and controlled cooling rates when placed in a -80°C freezer overnight.
Transfer the vials into liquid nitrogen for long-term storage

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.

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.

Our skeletal muscle cells are mainly isolated from M. pectoralis; sometimes also from M. gastrocnemius; M. intercostales or M. gluteus maximus. The exact localization is specified in the Certificate of Analysis.

Short protocol:

Wash the cells with sterile PBS 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.

A concentration > 10% FBS is needed to completely inactivate the trypsin. As most of PromoCell's growth media are serum-reduced or serum-free; the use of a trypsin inhibitor like TNS is highly recommended.

We recommend to use the trypsin as well as the other detach solutions at room temperature to avoid overtrypsinization and irreversible cellular damage.

Our TNS solution contains 0.05% (w/v) trypsin inhibitor from soy bean in HepesBSS/0.1% BSA.

The practice of heat inactivation was originally developed when only serum from adult animals was available. Adult serum contains high serum complement which may destroy cells under certain conditions. Heating serum (30 min 56°C) is intended to inactivate the complement. Today serum is often heat-inactivated without any evidence of beneficial effect. When using FCS (fetal calf serum) heat inactivation is not necessary for most cell lines or cell types. PromoCell does not use heat-inactivated FCS for the preparation of their growth media.

Choose your region

North America

Europe

Asia

Technical Library