Technical library - RESOURCES

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).

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.

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.

Our Normal Human Cells have been cultured and tested in our growth media and have adapted to these conditions. Using other media may yield unsatisfactory results due to suboptimal supplies of nutrients and growth factors. PromoCell can only guarantee good cell growth (as stated in the Certificate of Analysis) when the cells are grown in the recommended media.

The human MSC derived from bone marrow adipose tissue and umbilical cord matrix are from different origins but with comparable biological properties and function. Depending on the tissue of origin they may have a higher preference for differentiation into one particular cell type and a lower preference for another one but they all still retain the differentiation potential for the mesenchymal lineage.

For optimal cell growth the human CD34⁺ Progenitors (C-12921) are cultured in PromoCell Hematopoietic Progenitor Cell Expansion Medium XF (C-28021) supplemented with Cytokine Mix E (C-39890) or with any other cytokine cocktail suitable to achieve optimal cell expansion. Cytokine Mix E contains recombinant human TPO SCF flt3-ligand and IL-3. The strong expansion of CD34⁺ cells will persist for approx. 2 weeks. The expansion factor is usually between 75 and 200-fold.

During the isolation of our Human Mononuclear Cells (hMNC) we first of all pay attention to thoroughly discard the platelet-containing fraction before we aspirate the MNC containing-interphase of the Ficoll gradient. The harvested MNC fraction is then subjected to several washing steps to remove potential remaining platelets. Finally the cell preparations are verified by microscopy to be largely free of platelet contamination.

There is a strong variation from donor to donor and sometimes from donation to donation concerning the number of vials that can be produced. This is due to individual variances between the donors as some of them have more MNCs per ml of blood and others have less. Generally; the number of vials (each with 25 x 10⁶ cryopreserved cells) per lot ranges between 10 and 40.

Pericytes have been shown to differentiate e.g. into adipocytes osteoblasts chondrocytes fibroblasts/myofibroblasts vascular smooth muscle cells and phagocytes.

CD146 (MUC18) is a surface marker expressed on pericytes MSCs and endothelial cells from large vessels (but not microvascular endothelial cells). In combination with CD34 (a marker for endothelial and hematopoietic cells) pericytes can be characterized by FACS analysis as CD146⁺/CD34^-. The absence of CD34 expression makes sure that the cells are not of hematopoietic/endothelial origin. Further markers that have been described for pericytes are NG2 CD90 alpha-SMA and PDGFR-beta.

Our kidney cells are characterized by their epithelial cell morphology and by cytokeratin expression using a pan-cytokeratin antibody.

Our human pulmonary microvascular endothelial cells (C-12281) are sourced from the lung parenchyma with all large vessels being removed beforehand. Therefore most of the HPMEC originate from capillaries.