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’s the difference between visceral and subcutaneous HWP? When should I use which cell type?
PromoCell Preadipocytes are isolated from subcutaneous or visceral fat.
Subcutaneous fat is found just underneath the skin and it is not related to the obesity-linked diseases. Its accumulation represents the normal physiological buffer for excess energy intake. When the storage capacity of subcutaneous fat is exceeded or the generation of new adipocytes is impaired, visceral fat starts to accumulate. It is located in the abdomen and around internal organs (e.g. kidney, heart, or bladder) and it is linked to hypertension, diabetes, and cardiovascular disease.
Adipocytes from subcutaneous and visceral fat differ in several functions, like their response to insulin and other hormones or their lipolytic activity. It highly depends on the scientific problem being addressed, which of the two cell types are best suited for your experiments.Related Links and Documents
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What is the exact localization of Human Dermal Lymphatic Endothelial Cells (HDLEC)? What medium should I use to culture them?
PromoCell’s HDLEC are isolated from skin (dermis). When isolated from juvenile donors (C-12216), the exact localization is foreskin. When derived from adult donors (C-12217), the localization depends on the type of surgery, e.g. breast or temple. You can find the information on the exact localization in the Certificate of Analysis.
Our HDLEC are tested to be positive for CD31, podoplanin, and ac-LDL uptake and are delivered in P2. The recommended culture medium is Endothelial Cell Growth Medium MV2 (C-22022). -
How should I use accutase for cell detachment?
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.
Related Links and Documents
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What is the difference between DetachKit (C-41200) and DetachKit-2 (C-41202)?
Both Kits contain HepesBSS, trypsin/EDTA, and Trypsin Neutralizing Solution (TNS). In DetachKit (C-41200), the concentration of trypsin/EDTA is 0.04% / 0.03%. In DetachKit-2 (C-41202), it is reduced to 0.025% trypsin / 0.01% EDTA. HepesBSS and TNS concentrations are identical in both Kits.
Related Links and Documents
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How does PromoCell characterize their MSC-AT (C-12977)?
Our hMSC-AT are characterized by their differentiation potential into chondrocytes, fat cells, and bone cells. In addition, we determine the presence of CD73, CD90 and CD105 expression as well as the absence of CD14, CD19, CD34, CD45 & HLA-DR expression by flow cytometry as proposed by the International Society for Cellular Therapy.
Related Links and Documents
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Do I have to use coated tissue culture vessels for culturing PromoCell Normal Human Cells?
It is not necessary to use coated flasks for (most of) our Normal Human Cells but it can be done. As coating with extracellular matrix proteins can affect cellular metabolism, it is recommended to use the same coating material for a complete set of experiments. Cells that need to be grown on coated dishes:
- Mesenchymal Stem Cells (C-12974/C-12971/C-12977) need Fibronectin- or Vitronectin-coating when grown in PromoCell MSC Growth Medium XF (C-28019) and when differentiated in MSC Neurogenic (C-28015), Adipogenic (C-28016), or Osteogenic (C-28013) Differentiation Media.
- Human monocyte-derived macrophages (C-12914/C-12916/C-12915/C-12917) must be seeded into Fibronectin- or Vitronectin-coated culture vessels in combination with PromoCell’s M1- and M2-Generation Media XF (C-28055, C-28056).
- For efficient induction of osteoblast mineralization with PromoCell’s Osteoblast Mineralization Medium (C-27020), the TC plates should be pre-coated with collagen type I.
Related Links and Documents
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At what passage do PromoCell test the cell type-specific markers?
Cell type-specific markers are usually determined one passage after thawing.
Example: After thawing, HUVEC are in P1. Markers (CD31 expression, Dil-Ac-LDL uptake) are tested in P2.
Please note: Cells that are frozen directly after isolation (blood cells) are tested directly after thawing with no further culturing step. -
What is the definition of “adult stem cells”?
“Adult stem cells” are stem cells isolated from postnatal tissues that have retained the capacity for cell renewal as well as for differentiation into multiple lineages (multipotency).
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What is the time frame for the differentiation of HWP? Can PromoCell provide a differentiation protocol?
Induction of differentiation:
– Grow the cells in Preadipocyte Growth Medium (C-27410) until they reach complete confluence; this roughly takes 1 week. Change medium 2-3x per week.
– Aspirate the Growth Medium; add Preadipocyte Differentiation Medium (C-27437) for 72 h
– Aspirate the Differentiation Medium, add Adipocyte Nutrition Medium (C-27438).
The cells will now start to accumulate droplets of lipids which can be visualized under the microscope. The process usually takes 1-2 weeks. Change medium 2-3x per week. It is possible to maintain the differentiated adipocytes in Nutrition Medium for up to 4 weeks. However, the cells tend to lyse and detach after about 3 weeks.Please note: We recommend to perform differentiation experiments at population doubling numbers lower than 4-5 (a maximum of 1 passage after thawing) in order to obtain a high differentiation level of the culture.
Related Links and Documents
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The melanocytes from Promocell are primary cells, so how many passages can they go before losing activity?
PromoCell guarantee 15 population doublings for their NHEM. In terms of passages this corresponds to approx. 4-6 subcultivations. We can however not guarantee that the activity of all melanocyte genes remains unchanged during this time. Primary cells do gradually change their phenotype in vitro. Therefore, it is recommended to use the cells for the important experiments at low passages.
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