Technical Library

Recent Entries in Technical Library

• 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
• Is it possible to split differentiated adipocytes in order to perform more experiments?

  Generally, you can split differentiated adipocytes. But these cells lose their ability to proliferate after differentiation and you can’t expand the culture any more. Therefore it is recommended to plate the cells into the needed vessels (e.g. multiwell plates) prior to induction of differentiation so that trypsinization isn’t necessary.

  Related Links and Documents

  • HWP
• Can PromoCell supply melanocytes from asian donors?

  The majority of our skin tissue donors are caucasians. But occasionally we also get skin biopsies from asian and black donors.
  Please contact our Technical Customer Support if you need cells from a particular phototype or origin. They will check our inventory and send you a list of available cell lots.

  Related Links and Documents

  • Melanocyte cell culture
• How does PromoCell recommend subculturing the chondrocytes (e.g. from T25 into T75 flasks, or into petri dishes)? Does PromoCell recommend a specific type or brand?

  We recommend a seeding density for chondrocytes between 10,000 and 20,000 cells/cm². This means that a subconfluent T25-flask with approx. 900,000 cells/T25 flask (36,000 cells/cm² ) may be either split into 3 new T25 or seeded in one T75 flask or in one 100 mm petri dish. We do not recommend a specific type or brand for the culture of HCH.

  Related Links and Documents

  • HCH
• 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
• What calcium concentration is needed to induce differentiation in cultured keratinocytes?

  The optimal calcium concentration for both proliferation and differentiation of keratinocytes depends on the species and also on the media formulation. To keep primary human keratinocytes in the proliferative status, concentrations between 0.03 and 0.15 mM (PromoCell Keratinocyte Growth Medium 2: 0.06 mM) are generally used. Increasing the calcium above 1 mM will induce terminal differentiation and lead to the loss of proliferative activity.

  Related Links and Documents

  • Keratinocyte Growth Medium 2
  • NHEK
  • CaCl2 solution
• Why does PromoCell use CD146 as a marker for pericytes?

  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.

  Related Links and Documents

  • hPC-PL
• 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

  • hPC-PL
• What is the exact localization of PromoCell’s HRCEpC? Are the cells isolated from proximal or distal tubuli?

  Our Human Renal Cortical Epithelial Cells (C-12660) are isolated from the cortex of the human kidney. The renal cortex is the outer portion of the kidney. It contains the renal corpuscles, the proximal and distal convoluted tubules, and the cortical collecting ducts.

  Related Links and Documents

  • HRCEpC
  • Schematic structure of a kidney
• From what part of the lungs does PromoCell isolate the human pulmonary fibroblasts (HPF)?

  Our HPF (C-12360) are isolated from peripheral lung tissue.

  Related Links and Documents

  • HPF