Technical Library

Recent Entries in Technical Library

• Can PromoCell recommend a medium to co-culture endothelial cells and fibroblasts?

  Some of our customers have successfully used PromoCell Endothelial Cell Growth Medium MV2 (C-22022) when co-culturing Human Coronary Artery Endothelial Cells (HCAEC; C-12221) and Normal Human Dermal Fibroblasts (NHDF; C-12300).

  Related Links and Documents

  • Endothelial Cell Growth Medium MV2
• What are the advantages of using PromoCell DC Generation Medium XF for dendritic cell generation?

  Our DC Generation Medium XF (C-28052) has a xeno-free formulation. It provides a complete media system (ready-to-use, all cytokines included) and shows efficient and reproducible in vitro maturation of moDCs from freshly isolated peripheral blood monocytes.

  Related Links and Documents

  • Dendritic Cell Generation Medium XF
• How does PromoCell ensure that there is no fibroblast contamination in the pericytes? Are the fibroblasts removed by separation with anti-CD90 labeled magnetic beads?

  No, we don’t perform CD90 immunomagnetic separation with our pericytes. We can however exclude fibroblast contamination as follows:
  1) The presence of fibroblasts in our pericyte cultures would be detectable shortly after cell isolation, as pericytes need up to 2 weeks before they start proliferating. Fibroblasts on the other hand would proliferate immediately and overgrow the culture.
  2) Characterization of the isolated pericytes during QC includes flow cytometry of CD146. Pericytes express CD146 whereas placental fibroblasts don’t.

  Related Links and Documents

  • Human Pericytes from Placenta
• My cells didn’t detach during subculture. What could be the reason?

  Please find attached a trouble shooting guide to identify possible reasons for poor detachment during subculture.

  Related Links and Documents

  • Trouble shooting guide: Slow detachment
• Why is the growth rate of PromoCell’s HDMEC significantly reduced when I remove hydrocortisone from the Endothelial Cell Growth Medium MV Kit (C-22120)?

  Epidermal Growth Factor (EGF) and hydrocortisone have been reported to have a synergistic effect on the growth of microvascular endothelial cells. Accordingly, if you remove the hydrocortisone, the proliferation of HDMEC is clearly affected.

  Related Links and Documents

  • Endothelial Cell Growth Medium MV
• Should I warm the trypsin prior to use?

  We recommend to use the trypsin as well as the other detach solutions at room temperature to avoid overtrypsinization and irreversible cellular damage.

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  • DetachKit
• 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

  • DetachKit
• 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.

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  • hMSC-AT
• 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-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-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

  • Mesenchymal Stem Cell Culture
  • Macrophage Cell Culture
  • Osteoblast Cell Culture
• 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.