Have a question about one of our products? Check out our technical library for recently asked questions from other scientists around the world.
Technical Library
Recent Entries in Technical Library
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What are the key points relating to proliferation, differentiation and culturing of HWP?
Our Normal Human White Preadipocytes (HWP) are isolated from adult subcutaneous or visceral adipose tissue from different locations. The cells are frozen in Cryo-SFM at the end of passage 1 (= secondary culture). A randomly selected vial is then used for quality control, which includes determination of growth characteristics, control of morphology, and tests for differentiation capacity into mature adipocytes.The recommended seeding density of preadipocytes after thawing/trypsinization is 5,000 cells/cm2. The cells should be trypsinized before reaching 90% confluence.
Population doubling times are usually between 20-40 hrs (10 population doublings guaranteed). Using a 1:4 split ratio, you can perform ∼4-5 passages with the cells.Preadipocyte Growth Medium (C-27417) is used to propagate the cells. To induce differentiation of preadipocytes into adipocytes, cells are grown in PromoCell’s Preadipocyte Growth Medium until they reach 100% confluency. Cells are then cultured in Preadipocyte Differentiation Medium (C-27437) for 72 h, followed by 10-14 days in Adipocyte Nutrition Medium (C-27439). During this time the cells start to accumulate fat droplets which can be visualized under the microscope. We recommend performing differentiation experiments at population doubling numbers lower than 4-5, in order to reach a high differentiation level of the culture.Related Links and Documents
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Why is the serum content of PromoCell’s specialized media so low?
FCS is a natural product consisting of many different components like salts, hormones, vitamins, trace elements, proteins, and enzymes. There can be large lot-to-lot variations between different serum batches regarding the concentrations of growth-promoting factors. The higher the serum content in a culture medium, the higher the impact of the variations on the cell culture system. For this reason, PromoCell has developed several serum-reduced and serum-free media where part of the serum has been replaced by more defined factors like cytokines, hormones or vitamins.Media with reduced serum concentrations have the benefit that they produce more standardized culture conditions over a long time span.
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Does PromoCell determine the percentage of HDLEC in its HDMEC lots?
No, we don’t determine the ratio of HDLEC and HDBEC in our HDMEC lots. From our experience, the percentage of HDLEC is highly lot-dependent and can vary between 5 and 60%.
Related Links and Documents
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Can I use the Growth Medium instead of TNS to stop the trypsin when splitting the cells?
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.
Related Links and Documents
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What is the shelf life of the PromoCell DetachKit/DetachKit-2?
The three components of DetachKit and DetachKit-2 (HepesBSS, Trypsin/EDTA, TNS) are stable for 1 year from the date of manufacture when stored at -20°C. Once thawed for usage, they should be stored at 4-8°C and can be used for 6 weeks. Please avoid repeated freeze/thaw cycles.
Related Links and Documents
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I would like to know the number of population doublings for PromoCell mononuclear cells (C-12907/C-12901) when grown in Mononuclear Cell Medium (C-28030).
Mononuclear cells are mostly used in immunology, infection biology, hematology and cancer research to study subpopulations of blood cells.
Our Mononuclear Cell Medium (C-28030) is intended for short-term maintenance (up to 48 hrs) of the thawed hMNC before you proceed with your experiments. The number of PDs will depend on the subsequent cell culture conditions and is not determined by PromoCell.
Please note: Depending on the conditions, the ratio of the subpopulations will gradually change, as the different blood cell types behave in different manners. Researchers normally start soon after thawing to either select the cell type of their interest (e.g. hematopoietic cells, endothelial progenitor cells) or perform experiments with all populations of hMNCs (e.g. to study effects on toxicity, viability or metabolism).Related Links and Documents
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How can I calculate the population doubling time?
The population doubling time or generation time (tg) is usually calculated during the logarithmic phase of growth. It specifies the time (t) in hours needed by the culture to double its cell number.
tg = t / nn: number of population doublings
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What is the precise localization for a) HNEpC, b) HTEpC, c) HBEpC, and d) HSAEpC?
a) HNEpC (Human Nasal Epithelial Cells) are isolated from nasal mucosa
b) HTEpC (Human Tracheal Epithelial Cells) from the surface epithelium of trachea
c) HBEpC from the surface epithelium of bronchie, and
d) HSAEpC (Human Small Airway Epithelial Cells) from the distal portion of the respiratory tract in the 1 mm bronchiole area
(comprising the cells from bronchioli and alveoli).Related Links and Documents
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What is the difference between “primary cells” and “normal cells”?
Freshly isolated cells that are plated in a tissue culture vessel for the first time are named primary cells or primary culture (corresponding to P0). As soon as they have been subcultured, they should correctly be termed normal cells (> P1).
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Is everything in the PromoCell Skeletal Muscle Cell Differentiation Medium (C-23061) defined?
Yes, our Skeletal Muscle Cell Differentiation Medium is completely defined. The SupplementMix (C-39366) consists of recombinant human insulin.
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