Technical Library

Recent Entries in Technical Library

• Where can I find the composition of the supplements?

  The qualitative and quantitative composition of the supplements can be found on our website and in the data sheets of the specialized media. When there is no such information specified, the composition of the supplements is confidential.

• 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
• What is the difference between juvenile (C-12210) and adult HDMEC (C-12212)? Which ones should I use for my experiments?

  Our juvenile HDMEC (C-12210) are isolated from foreskin of young male donors (1-10 years). In contrast, adult HDMEC (C-12212) are derived from different skin localisations like the cheek, temple, or breast. The donors are > 20 years old and are mostly female.
  Adult HDMEC are the cells of choice when you need cells from a particular part of the body (other than foreskin), or if it is important for your study to use cells from female and/or adult donors.

  Related Links and Documents

  • HDMEC
• Is it necessary to use the PromoCell DetachKit when subculturing PromoCell Normal Human Cells?

  We recommend to use the DetachKit when subculturing our Normal Human Cells. It contains a HepesBSS washing buffer, trypsin 0.04% / EDTA 0.03% solution, and TNS, a trypsin inhibitor from soybean.
  Please note: Many of our culture media have low serum content or no serum at all. These media are not suitable to inactivate trypsin during subculture.

  Related Links and Documents

  • DetachKit
  • Subcultivation of primary cells
• I cannot find the Certificate of Analysis (CoA) pertaining to my cells. Can you please send me a copy?

  The Certificates of Analysis can easily be downloaded from our PromoCell website:
  https://www.promocell.com/certificate-of-analysis/
  Simply type in the lot number indicated on the cryovial/TC-flask and click the SEARCH button.

  Related Links and Documents

  • Download CoA from website
• How can I successfully isolate HUVEC from umbilical cords without using antibiotics?

  At PromoCell, we get the umbilical cords from our tissue suppliers with no addition of buffers or media. This method prevents the microorganisms from being washed into the blood vessels. Before we start the cell preparation, the umbilical cord is also cut on both ends with a sterile scalpel to provide sterile intersections in addition to the sterile lumen.
  This method allows us to isolate sterile endothelial cells from umbilical vein and to plate them in antibiotics-free culture media.

  Related Links and Documents

  • HUVEC
• 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
• What culture conditions are required for culturing PromoCell Normal Human Cells in the respective PromoCell media?

  PromoCell Normal Human Cells should be cultured in the appropriate medium at 37°C and 5% CO₂ in a humidified atmosphere.
  Please note: If using cell culture flasks w/o filter cap, unscrew the cap by half a turn to allow sufficient ventilation.

• Can you please provide me a protocol for the expansion of CD34+ Progenitor Cells in PromoCell Expansion Medium XF?

  Short protocol:

  • Thaw the cells (C-12921) for 2 min in a 37°C waterbath. Dilute in 9 ml of complete HPC Expansion Medium XF (+ Cytokine Mix E) and count the cells
  • Spin down for 10 min at 240xg, aspirate the supernatant, resuspend the pellet at 20,000 cells/ml HPC Expansion Medium XF
  • Plate in an appropriate suspension culture vessel and incubate the culture for 2-3 days at 37°C and 5% CO₂
  • Then double the media volume by adding fresh complete medium, e.g., 4 ml  suspension culture + 4 ml fresh medium (= 8 ml)
  • Incubate the cells for an additional 10-12 days by performing a partial medium change every 2-3 days
    Example partial medium change: For a culture volume of 8 ml, spin down the cells, aspirate and discard 4 ml of the supernatant, resuspend the cells and add 12 ml of fresh complete medium (= 16 ml).

  In combination with the Cytokine Mix E, the HPC Expansion Medium XF typically promotes a 300-1,000 fold expansion of the total cell population. After 2 weeks of expansion about 20-30% of the population express CD34⁺, indicating a 50-200 fold expansion of CD34⁺ progenitor cells.

  Related Links and Documents

  • App Note: Expansion of primitive Hematopoietic Progenitor Cells
  • Hematopoietic Stem Cell Culture
• Why does the macrophage differentiation in the PromoCell protocol take 10 days? In the literature or following the feedback from other researchers, it usually only takes 7 days.

  M1 / M2 polarization also takes seven days in the PromoCell system but the protocol contains two more days for optional macrophage activation. If you only want non-activated M1 / M2 macrophages, the process is usually completed after 7 days.
  Nevertheless, PromoCell does not recommend shortening the 10-day protocol because you actually get a plus in viability and cell yield (due to the re-attachment of floating cells) on day 8-10 due to the media change.

  Related Links and Documents

  • Application Note: Differentiation of M1- or M2-Macrophages
  • M1-Macrophage Generation Medium DXF
  • M2-Macrophage Generation Medium DXF