Technical Library

Recent Entries in Technical Library

• What is the difference between “primary cells” and “normal cells”?

  Freshly isolated cells that are plated in a tissue culture vessel for the first time are named primary cells or primary culture (corresponding to P0). As soon as they have been subcultured, they should correctly be termed normal cells (> P1).

• 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-27436) 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

  • HWP
  • Preadipocyte/Adipocyte Media
  • HWP and related media
• 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.

  Related Links and Documents

  • Melanocyte Cell Culture
• Where does PromoCell source the heart tissue for HCM isolation?

  The tissue comes from patients who underwent heart transplantation. We obtain a part of the explanted (not the transplanted) heart to prepare cardiac myocytes.

  Related Links and Documents

  • Human Cardiac Myocytes
• I received 3 DetachKits yesterday. Some of the tubes have different colors, although they all come from the same batch.

  The components of the PromoCell DetachKit may occasionally arrive with a non-uniform color appearance. This phenomenon is known to PromoCell’s Quality Assurance Dept.
  It is reversible and has no influence on the quality of the product.

  Related Links and Documents

  • PromoCello DetachKit
• Will the NHEM.f (C-12400) and NHEM.f M2 (C-12402) also grow in PromoCell Melanocyte Growth Medium M3?

  Yes, NHEM.f and NHEM.f M2 also grow in our optimized Melanocyte Growth Medium M3 (C24310) and can achieve > 15 population doublings.

  Related Links and Documents

  • Melanocyte Cell Culture
• Why is the actual cell count in the vial at PromoCell sometimes significantly higher than the 500,000 cells stated on the website?

  We guarantee > 500,000 viable cells per vial. To do this, we need to freeze more than 500,000 cells, as we do not yet know the viability after thawing at the time of freezing.
  For technical/organizational reasons, the initial cell number may also vary from lot to lot, so 2 lots with the same viability may not necessarily contain the same number of cells.

  Related Links and Documents

  • Human primary cell culture
• Has PromoCell done any tests with MSC Adipogenic Differentiation Medium 2 without fibronectin? Is fibronectin coating really necessary?

  The MSCs attach even without fibronectin coating, but the risk of detachment during differentiation is much higher. We therefore strongly recommend using fibronectin-coated TC vessels.

  Related Links and Documents

  • Human Mesenchymal Stem Cells
  • Mesenchymal Stem Cell Adipogenic Differentiation Medium 2
• I would like to know if the Lymphocyte Separation Medium (C-44010) is endotoxin tested.

  Yes, our Lymphocyte Separation Medium 1077 is endotoxin-tested. The specification for the product is < 10EU/ml.

  Related Links and Documents

  • Lymphocyte Separation Medium
• Is the PromoCell Keratinocyte Growth Medium 2 suitable for growth of primary mouse keratinocytes?

  Yes, primary mouse keratinocytes grow well in this media when you reduce the CaCl₂ concentration to 0.025 mM – 0.05 mM CaCl₂ (optimal Ca concentration for primary human keratinocytes is 0.06 mM).

  Related Links and Documents

  • Keratinocyte Growth Medium 2