Cells and Media
What is the definition of “normal human cells”?
“Normal” means that the cells in question were freshly isolated from non-tumour tissue. They have a finite life span in vitro. They have neither been genetically modified nor treated and/or selected to produce a continous cell line. “Normal” cells are primary cells that have been subcultured at least once.
What is the difference between “population doublings” (PD) and “passage”?
A population doubling (PD) is a two-fold increase in the total number of cells during in vitro-culture. Calculating the number of PDs a culture has performed at a certain point gives an accurate measure of its age. In contrast, the term passage merely refers to the subculturing process, i.e. the detachment of the cells from a culture vessel (using eg. trypsin/EDTA or accutase) and replating them at a lower density to allow for further cell growth. The term passage does not take into account the different split ratios that can be applied. Different split ratios produce cultures with an identical number of passages but a varying number of doublings. Passage is therefore only a rough estimate of the actual age of a culture.
Can I store the cryopreserved cells at -80°C?
Storage at – 80°C is not sufficient for long-term preservation of primary cells and causes irreversible cell damage. Please store them in liquid nitrogen (< -150°C).
Does PromoCell have an informed consent available for every tissue they use for cell isolation?
Yes, all tissues used by PromoCell for the isolation of human cells are derived from donors who have signed an informed consent form (either by the donor himself, an authorized agent, or a legal agent). When obtaining and using human tissue, PromoCell acts in strict compliance with “The Convention for Protection of Human Rights and Dignity, “The Declaration of Helsinki”, “The Human Tissue Act”, and “The German Federal Data Protection Act”.
Are PromoCell Cells derived from single donors or are they pooled from multiple donors?
Unless otherwise stated, PromoCell Human Cells are derived from single donors. The donor’s age, sex, and race are specified in the lot-specific Certificate of Analysis which can be downloaded here. For a few selected cell types, we also supply cells from pooled donors. These include HUVEC pooled (# C-12203/C-12253), HUVEC 2 pooled (# C-12208/C-12209) and NHEK pooled (# C-12005/C-12007; C-12006/C-12008).
What culture conditions are required for culturing PromoCell Normal Human Cells?
PromoCell Normal Human Cells should be cultured 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 for sufficient ventilation.
Can I grow PromoCell Normal Human Cells with antibiotics?
For optimal cell growth, we recommend to refrain from using antibiotics. Most primary or normal human cells show reduced growth rates in the presence of antibiotics. In some cases, when a sterile environment cannot be 100% ensured, it may be advisable to add antibiotics to the media to protect the culture from potential microbial infections.
At what passage should I perform my experiments when working with PromoCell Normal Human Cells?
There is no general recommendation regarding the passage number at which to perform your experiments. This depends on the cell type as well as on the type of experiment. Most scientists don’t exceed passage 4 to 6 when using normal cells for their experiments as the cells gradually change their phenotype in vitro. If a certain phenotype is essential for your experiments, you should run expression controls at regular intervals to ensure that the markers are still sufficiently expressed in your cells.
What is the difference between cryopreserved and the proliferating cells?
Our cryopreserved cells are frozen in Cryo-SFM (# C-29910) and are shipped on dry ice. After delivery, they must be transferred into liquid nitrogen or they can be thawed and directly seeded into an appropriate tissue culture vessel using the recommended Growth Media. Proliferating cells are thawed at PromoCell and shipped as growing cultures in T25 flasks (T75 for Pericytes).
What is the recommended split ratio for PromoCell Normal Human Cells?
PromoCell does not recommend a split ratio for Normal Human Cells because the cell yield varies from lab to lab. We recommend to count the cells after trypsinization and to replate them at 5,000-10,000 cells/cm². The precise recommended plating density for each cell type can be found in the Instruction Manual.
What markers does PromoCell test to characterize their Normal Human Cells?
For specific marker details, please see the Instruction Manual of each respective cell type or view the Certificate of Analysis (available for download online for each lot).
What is the shelf life of the PromoCell media and supplements?
The shelf life of the majority of our basal media is 9 months from the date of manufacture (when stored between 4°C – 8°C). The shelf life of the supplements is 12 months from the date of manufacture (when stored at -20°C). The exact expiry date is specified on the product label.
How long can I use the supplemented media?
Once you have added the supplements to the basal medium, the shelf life is 6 weeks at 4°C (with a few exceptions). The stability of the complete media is also specified in the respective Instruction Manual (“Storage and Stability”). Please avoid repeated heating of the media to 37°C as this will reduce the activity of the growth factors and the concentration of L-glutamine.
Can I use the PromoCell growth media for primary cells of other species?
The PromoCell media are optimized for in vitro-culture of human cells. In most cases, they can also be used for cells from several other species (eg. rat, mouse, bovine, porcine cells). To view a list of references where our media were used with other species/cell types please click here.
What is the difference between PromoCell Medium “ready-to-use” and PromoCell Medium Kit?
PromoCell Cell Culture Media “ready-to-use” consist of basal medium and SupplementMix. PromoCell Culture Media Kits consist of basal medium and SupplementPack. Addition of the supplements (SupplementMix or SupplementPack, respectively) to the appropriate basal medium will result in identical growth media. For additional information please click here.
What is the difference between SupplementMix and SupplementPack?
In the PromoCell SupplementMix, all growth supplements are pre-mixed in one tube. In the SupplementPack, all growth supplements come in separate vials. For additional information please click here.
Where can I download the Certificate of Analysis of my cells?
The CoAs can be downloaded here. Simply type in the lot number and click Search.
How can I place my order?
To place an order, you can use any of the following options: fax, email or our online shop. If you wish to order by fax, please click here to select your country and get the corresponding fax numbers. To order via email, please send your order to firstname.lastname@example.org. If you wish to order via our online shop, you first need to create an account and subsequently easily log-in for all future orders.
What information does PromoCell need to properly process my order?
The following information is necessary to correctly process your order: Shipping and billing address, contact person’s name and phone number, catalog number(s), size, and quantity. Please also clearly indicate if you have a customer number, reservation number (ATxxxxxx), or a quotation (ANxxxxx).
Do I receive an order confirmation?
When you order via our online ordering system, you will get an automatic order confirmation email. If you order by email or fax, you will not receive an order confirmation.
Are there special shipping dates for PromoCell proliferating cells?
Our proliferating cultures are generally dispatched on Mondays (delivery on Tuesday) and on Thursdays (delivery on Friday). To the US and Canada, we only ship proliferating cells on Mondays.
Please note: Proliferating cells are only sent out to destinations where we can guarantee express delivery within 24 hrs.
Where do the primary human cells come from? Are they able to endure such a long trip?
All cells are shipped from our premises in Germany. The cryopreserved vials are shipped with plenty of dry ice. For some countries, when delivery cannot be guaranteed within 3-4 days, we use vapor shippers. Upon receipt of the cells, store them in liquid nitrogen until they are thawed and seeded. We have shipped many cryopreserved cells to countries all over the world. When they are thawed according to the PromoCell protocol, viabilities are achieved as specified in the CoA (≥70%).
How does PromoCell ship the proliferating cells?
Proliferating cells are shipped in T25 or T75 flasks completely filled with Growth Medium together with warm packs (37°C) that keeps the cells warm for several hours during the transport. Usually, the flasks arrive at room temperature and should be placed in the incubator for 3 hrs before changing the media or subculturing the cells.
Please note: Proliferating cultures are only shipped to those countries where a 24 h delivery can be guaranteed.
To which countries does PromoCell supply its products?
We deliver directly to Germany, France, Monaco, the United Kingdom, the United States and Canada. In addition, we have distributors in more than 40 countries worldwide. You will find current information on PromoCell distributors here.
What shipping costs does PromoCell charge?
We charge the following shipping costs, irrespective of the total amount of your order:
Germany/ France/ Monaco: 32 € / United Kingdom: £26 / USA/Canada: $59 / Rest of the world: Please inquire!