Technical Library

Recent Entries in Technical Library

• 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

  • HDMEC
• 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

  • Trypsin Neutralizing Solution (TNS)
• When should I use DetachKit, when DetachKit-2?

  Our DetachKit (C-41200) is optimized for primary cells and can be used to trypsinize all Normal Human Cells supplied by PromoCell.
  Detach Kit-2 was developed for very gentle detachment and is still used by some investigators to subculture Human Nasal Epithelial Cells (HNEpC).

  Related Links and Documents

  • DetachKit
• Can I detect the differentiation of Mesenchymal Stem Cells by their morphology, without doing any cell staining?
  • Adipogenic differentiation can be identified morphologically and without any staining by the formation of intracellular lipid vesicles.
  • In contrast, when MSC differentiate into bone cells, there is no significant change in morphology. It is recommended to perform Alkaline Phosphatase staining to detect osteoblastic differentiation or Alizarin Red S staining to show osteoblast mineralization.
  • Chondrogenic differentiation is generally performed as spheroids in 3-D cell culture and not in 2-D monolayer culture. Staining with Alcian Blue to visualize the differentiation process is indispensable.
  • Neurogenic differentiation can be detected using neuron specific markers (e.g. beta-3 tubulin, NeuN, MAP2) and by their typical neuronal morphology.

  For differentiation protocols, please see attached Application Notes.

  Related Links and Documents

  • Product Manual: hMSC
  • Application Note: Adipogenic Differentiation and Analysis of MSC
  • Application Note: Osteogenic Differentiation and Mineralization of MSC
  • Application Note: Chondrogenic Differentiation and Analysis of MSC
  • Application Note: Neurogenic Differentiation and Analysis of MSC
• What markers does PromoCell test to characterize their Adult Human Stem and Blood Cells?

  All Adult Human Blood and Stem cells are analysed by flow cytometry to express defined markers. MSCs are additionally tested for their capacity to differentiate into the osteogenic, chondrogenic and adipogenic lineages. For more details, please see Certificate of Analysis of the respective cell type (“Phenotypic characterization”).

  Related Links and Documents

  • Human Stem & Blood Cell Culture
• Can the cell pellets be used to establish a growing culture?

  No, the PromoCell cell pellets (C-14**) are frozen at -20°C and cannot be revived. Their main application is to analyze RNA or protein. Cryopreserved cells that can be revived are available from the same donors. Please contact our Technical Customer Support if you need matched viable cells.

• How many passages can I perform with PromoCell Normal Human Cells?

  PromoCell does not determine the number of passages but instead we calculate the population doublings (PD) that can be performed with the cells. The term passage only describes the process of detachment and replating and does not take into account different split ratios. The optimal split ratio is calculated from the actual cell yield after trypsinisation and the recommended plating density. In most of our cell types, the split ratio is usually between 1:3 and 1:6. Using 1:4 splits (i.e. increasing the growth surface by factor 4 each time), 15 doublings are achieved after 6-8 passages.
  For recommended plating densities, please view the respective Manual, section “Specifications”.

• Can I first expand the osteoblasts and then perform my experiments?

  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.

  Related Links and Documents

  • HOB
• Does the Chondrocyte SupplementMix (C-39635) contain any growth factors?

  The Chondrocyte Growth Medium SupplementMix consists of Fetal Calf Serum (final concentration 10 % v/v) which has specifically been tested for the culture of primary chondrocytes. FCS contains a variety of different growth factors, which are however, not analyzed in more detail. There are no further growth factors added to the SupplementMix.

  Related Links and Documents

  • Chondrocyte Growth Medium
• Where does PromoCell source the bronchial tissue for HBEpC preparation? Are the smoking habits of the donors known?

  We source the bronchial tissue from forensic medicine and from thoracic surgery. For some lots, the smoking habits of the donors are known.
  Please contact our Technical Customer Service before ordering the cells if you need this information.

  Related Links and Documents

  • HBEpC