Technical library

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.

PromoCell Cell Culture Media "ready-to-use" consist of basal medium and SupplementMix. PromoCell Culture Media Kits consist of basal medium and SupplementPack. Addition of the supplements (SupplementMix or SupplementPack, respectively) to the appropriate basal medium will result in identical growth media.

The medium supplemented with Cytokine Mix E is stable for 2 weeks if stored protected from light at 2-8°C.

Short description:

Unpack the box and place the T25 flask(s) in the incubator for 3 hrs (closed cap). Then check confluency under the microscope.

When the density is < 70%, aspirate the medium using sterile conditions and add 5-10 ml of the appropriate Growth Medium.

The cells should be subcultured according to the subcultivation protocol given in the cells' Instruction Manual once they have reached > 70 % confluency.

It usually takes 5 to 8 days to grow our Normal Human Chondrocytes (C-12710) to subconfluency. The number of doublings (PDs) they undergo can be calculated from the number of seeded cells and the cell yield at subconfluency. Generally, when HCH are plated with 10,000 cells/cm² they perform between 1.5 and 2 doublings per passage.

Our subcutaneous HWP are isolated from subcutaneous fat of different localizations, e.g. abdomen, breast, or upper arm. The visceral preadipocytes are isolated from fat surrounding e.g. the pericardium, or from the omentum or mediastinum. The exact localization is specified in the Certificate of Analysis. If you need HWP from a particular localization, please contact our Technical Customer Support prior to placing your order.

The time needed to detach our primary cells depends on many different factors like the cell type, cell density, lot #, trypsin concentration, the efficiency of the washing step before adding the trypsin and the trypsinization temperature. For most cell types we recommend trypsinization at room temperature and direct observation of detachment under the microscope. This way, you can find out your individual trypsinization time and keep the contact time between cells and trypsin to a minimum. Most cells detach after 2-8 min. Please refer to the instructions in the Manual. For some cell types, trypsinization at 37°C or the use of Accutase or another Detachment Solution is recommended.

1) For fluorescence detection (fluorometer): Black plates with clear bottoms; often clear plates will suffice 2) For luminescence detection (luminometer): White/opaque plates 3) For colorimetric detection (photometer): Clear plates

NHEK.f are isolated from tissue samples (foreskin) of single donors, aged between 1-10 years. NHEK.f pooled are prepared from the foreskins of 3 individual donors. The cells of each donor are expanded in separate TC vessels and the cells are pooled after secondary culture, before cryopreservation. Both NHEK from single donor and NHEK pooled are in P2 after thawing.