Technical Library

Recent Entries in Technical Library

• Are endothelial cells α-SMA-negative under all circumstances?

  Scientific findings from different groups, as well as our own results indicate that the presence or absence of α-SMA is not a valid indicator for the composition of a given cell population. Research findings and our inhouse data suggest that endothelial cells are not α-SMA-negative under all circumstances. α-SMA negativity is not an intrinsic property of endothelial cells but can vary depending on extrinsic influences.

  Lot-specific values for α-SMA in our Certificates of Analysis (CoAs) are therefore not considered meaningful.

  Related Links and Documents

  • Endothelial Cell Culture
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  • Lu et al.; Stem Cells Dev . 2004 Oct;13(5):521-7
  • Azuma et al.; Biochem Biophys Res Commun . 2009 Mar 13;380(3):620-6
  • Frid et al.; Circ Res . 2002 Jun 14;90(11):1189-96
  • Kovacic et al.; J Am Coll Cardiol . 2019 Jan 22;73(2):190-209
  • Jones et al.; Am J Respir Cell Mol Biol . 1999 Apr;20(4):582-94
  • Ma et al.; Front Cell Dev Biol . 2020 Apr 21;8:260
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  • Amberger et al.; FEBS Lett . 1991 Aug 5;287(1-2):223-5
• Is vWF a great marker for Endothelial Cells?

  Secretory granules called Weibel-Palade bodies (WPBs) containing Von Willebrand factor (vWF) are more linked to the formation of a confluent endothelial monolayer. Research shows that vWF expression is dynamic and highly dependent on the cell culture conditions such as confluence and passage number.
  Therefore, having vWF as a quality control marker for each lot of ECs is not really necessary.

  Related Links and Documents

  • Endothelial Cell Culture
  • Howell et al.; Mol Membr Biol. Nov-Dec 2004;21(6):413-21
• The components of my DetachKit show different colors. Is this normal or does it affect their quality?

  The components of the PromoCell DetachKit may arrive on occasion with a non-uniform color appearance. This phenomenon is known by PromoCell’s Quality Assurance. It is reversible and does not influence the quality of the product.

  Related Links and Documents

  • PromoCell DetachKit
  • PromoCell DetachKit 2
• Can I defrost Accutase Solution, prepare aliquots and refreeze them?

  Yes, Accutase Solution can be defrosted, aliquoted, and then refrozen.

  Defrosting: Accutase should be defrosted overnight in the refrigerator or placed in a tub of cold tap water.  Do not defrost in a 37°C water bath.
  Stability: Once thawed, it is stable for at least 2 months in the refrigerator if stored promptly after use.

  Related Links and Documents

  • Accutase Solution
• Is it possible to refreeze the hCD34 progenitor cells after having amplifed them in PromoCell Hemaotopoietic Progenitor Cell Expansion Medium XF?

  Yes, it is possible to refreeze them.

  Related Links and Documents

  • CD34+ Progenitor Cells
  • HPC Expansion Medium XF
• Why are the cryopreserved macrophages abbreviated as hMDM-GMCSF(-) and hMDM-MCSF(-), respectively? What does the (-) stand for?
  • hMDM-GMCSF is the abbreviation for human monocyte-derived macrophages. They are polarized [⇒ differentiated with GM-CSF] but non-activated [⇒ (-)] M1 macrophages. 
  • hMDM-MCSF is the abbreviation for human monocyte-derived polarized [⇒ differentiated with M-CSF] but non-activated [⇒ (-)] M2 macrophages. 

  The macrophages can be seeded into all kinds of TC vessels. After plating, they can be maintained as biologically functional adherent cultures for several weeks.Optionally, user-customizable activation of the cells can be performed. For details, please see attached Application Note, page 2, Fig. 3 and page 5, Tab. 1).

  Related Links and Documents

  • Application Note: Assay-ready adherent cultures of fully functional human primary macrophages using cryopreserved cells
  • Cryopreserved Macrophages
  • Macrophage Generation Media DXF
• If I isolate fresh CD14-monocytes for M1/M2 macrophage generation, can I culture them for a period of time before differentiation?

  This is not advised. Please seed the freshly isolated CD14-monocytes immediately in the Monocyte Attachment Medium. Adding a culturing step will change the biological characteristics of monocytes very rapidly.

  Related Links and Documents

  • Application Note: Differentiation of M1- or M2-Macrophages from PBMC/Monocytes
  • Monocyte Attachment Medium
• Can you provide a list of cell lines that have been successfully used for tumorsphere formation with the PromoCell 3D Tumorsphere Medium XF?

  The following cell lines have been tested at PromoCell to form tumorspheres in 3D Tumorsphere Medium XF (C-28070):

  • U-87 MG
  • MCF-7
  • MDA-MB-231
  • HT-29
  • HT1080
  • HepG2
  • A-549
  • Panc-1
  • LNCaP
  • A-431

  ⇒ For more details, please view the attached Application Note.
  In addition, we have received customer feedbacks for the following cell lines:

  • HCT-116 (human colorectal carcinoma cell line)
  • Capan-1 (human pancreatic adenocarcinoma cell line)
  • PC3 (human prostate cancer cell line)
  • C42B (osteotropic prostate cancer cell line)
  • NCI-H23 (human lung epithelial adenocarcinoma cells)
  • IMR-32 (human neuroblast cell line)
  • A818-6 (human pancreatic ductal adenocarcinoma cell line)
  • HEK293 (human embryonic kidney cells)
  • Calu-1 (non-small-cell lung cancer cell line)

  Related Links and Documents

  • Application Note: Tumorsphere Culture of Cancer Stem Cells (CSC) with the PromoCell 3D Tumorsphere Medium XF
  • 3D Tumorsphere Medium XF
• Can PromoCell MSC Growth Medium 2 (C-28009) be used for Mesenchymal Stem Cells from rat?

  Yes, we have received a customer feedback that our MSC Growth Medium 2 also works for rat MSCs.
  The rat cells grow nicely in this medium and have a good viability.

  Related Links and Documents

  • MSC Growth Medium 2
• Which medium does PromoCell recommend for endothelial cell transfection ?

  There are several factors that can influence successful transfections, e.g. viability and density of the cells, choice of the transfection reagent, quality and type of the transfected molecules (plasmids, siRNA, oligonucleotides), as well as the culture medium and supplements used.
  When using PromoCell Endothelial Cell Growth Media for cell transfection, please follow the instructions below:

  Heparin, which is included in our Endothelial Cell Growth Medium (C-22010/C-22110), Medium 2 (C-22011/C-22111) and Medium MV (C-22020/C-22120) may reduce the transfection efficiency.  We therefore recommend to use our heparin-free Endothelial Cell Growth Medium MV2 (C-22022/C-22121). Alternatively, you may use Endothelial Cell Growth Medium Kit (C-22110), Medium 2 Kit (C-22111), or Medium MV Kit (C-22120) without adding the ECGS/heparin supplement to the Basal Medium.

  Please note: Before and after transfection, the cells should be cultured in complete Growth Medium including heparin to ensure optimal growth.

  Related Links and Documents

  • Manual: Endothelial Cell Growth Medium/Medium 2
  • Manual: Endothelial Cell Growth Medium MV/ MV2
  • Cell Transfection reagents