Technical Library

Recent Entries in Technical Library

• What is the exact localization of Human Dermal Lymphatic Endothelial Cells (HDLEC)? What medium should I use to culture them?

  PromoCell’s HDLEC are isolated from skin (dermis). When isolated from juvenile donors (C-12216), the exact localization is foreskin. When derived from adult donors (C-12217), the localization depends on the type of surgery, e.g. breast or temple. You can find the information on the exact localization in the Certificate of Analysis.
  Our HDLEC are tested to be positive for CD31, podoplanin, and Prox-1 and are delivered in P2. The recommended culture medium is Endothelial Cell Growth Medium MV2 (C-22022).

  Related Links and Documents

  • HDLEC
  • Endothelial Cell Growth Medium MV2
• How should I use accutase for cell detachment?

  Short protocol:

  • Wash the cells with sterile PBS or HepesBSS
  • Add undiluted accutase to the culture vessel (2 ml per 25 cm²)
  • Incubate at room temperature for 5-15 min or at 37°C for faster detachment
  • When the majority of the cells has detached, centrifuge the suspension and resuspend the pellet in fresh medium. In most cases no additional washes or neutralization steps are required.

  Related Links and Documents

  • Accutase Solution
• When should I order the Growth Medium Kit instead of the Medium “ready-to-use”?

  The Growth Medium Kit allows customization of the end concentrations of growth supplements. It is therefore more flexible than the Medium “ready-to-use” and can eg. be used to prepare a starvation medium.
  Please note: Modification of supplement concentrations may have an impact on cell growth. You should test in advance whether and for how long the cells can survive the altered culture conditions.

  Related Links and Documents

  • PromoCell Growth Medium Info Sheet
• I would like to know how homogenous the MSC population is and if all cells become osteoblasts when grown in MSC Osteogenic Differentiation Medium (C-28013).

  Human Mesenchymal Stem Cells have retained the ability to differentiate into a variety of cell types including fat cells, chondrocytes, and osteoblasts. PromoCell supply hMSC from 3 different tissues: bone marrow (hMSC-BM; C-12974), umbilical cord matrix (hMSC-UC; C-12971), and adipose tissue (hMSC-AT; C-12977).

  • MSC-BM show very good differentiation into bone cells but also into chondrocytes and fat cells when the respective Differentiation Media is used.
  • MSC-UC have a high potential to differentiate into chondrocytes but only weak potential for fat or bone cell differentiation.
  • In contrast, MSC-AT differentiate very well into fat and bone cells but only moderately into chondrocytes.

  In other words, to obtain a high percentage of bone cells, MSC-BM or MSC-AT are the cells of choice. There are of course lot-to-lot variations and the differentiation will decrease from passage to passage. If you need MSCs with a particularly high osteoblast differentiation capacity, you can call our Technical Customer Service before placing your order so that we can select an appropriate cell lot for you.

  Related Links and Documents

  • hMSC
  • MSC Differentiation Media
  • Application Note: Osteogenic Differentiation and Analysis of MSC
• Can PromoCell normal human cells be grown in standard media like DMEM or RPMI1640 supplemented with 10% FBS?

  PromoCell Growth Media have special formulations and are much more complex than DMEM or RPMI + FBS. In comparison to immortalized cell lines, primary cells have much higher nutrient and growth factor requirements. Classical media do not usually achieve good performances with our cells.

• Can I isolate RNA and/or proteins from the cell pellets?

  PromoCell cell pellets (C-14**) are an easily accessible source of DNA, RNA, and proteins.

• How many population doublings do PromoCell guarantee for the Normal Human Cells?

  PromoCell guarantee 15 population doublings (PD) for most Normal Human Cells (unless otherwise indicated on the Certificate of Analysis) when the recommended PromoCell media and the PromoCell DetachKit are used. For more information, please check the respective Manual, section “Specifications”.

  Related Links and Documents

  • Overview human primary cells
• How many osteoblasts will be in the flask when I purchase a proliferating culture (C-12760)?

  If you purchase a proliferating culture of human osteoblasts (C-12760), there will be > 500,000 cells in the TC-flask. Once the culture is subconfluent, you will count between 750,000 – 1.1 million cells/T25 (depending on the cell lot).

  Related Links and Documents

  • HOB
• Typically once seeded, how long does it take to grow the chondrocytes to subconfluence? How many doublings will they undergo per passage?

  It usually takes 5 to 8 days to grow our Normal Human Chondrocytes (C-12710) to subconfluency. The number of doublings (PDs) they undergo can be calculated from the number of seeded cells and the cell yield at subconfluency. Generally, when HCH are plated with 10,000 cells/cm² they perform between 1.5 and 2 doublings per passage.

  Related Links and Documents

  • HCH
• Is it necessary to cultivate Nasal Epithelial Cells on coated culture flasks?

  No, it is not necessary to use coated flasks, therefore we don´t recommend their usage in the Instruction Manual. However, for special applications, some of our customers use collagen-coated dishes.

  Related Links and Documents

  • HNEpC