For cell culture queries, please contact Dr. Ute Liegibel Product Specialist Cell Culture

Dr. Susanne Voltmer-Irsch Product Specialist Cell Culture

tech.supportpromocell.com

Answers

Can I freeze PromoCell cell culture media for long-term storage

Do not freeze PromoCell cell culture media. Freezing leads to irreversible precipitation of medium components.

How should I handle proliferating cells upon receipt from PromoCell?

Abridged protocol: Take the proliferating cells out of the shipping container and place them in the incubator for 2-4 hours without removing the transport medium. After this period, place the flask in a laminar flow hood, open the lid, and replace the transport medium with fresh cell culture medium. Cells are ready for subculture after reaching 90% confluence.

For detailed information see: 1.2 Handling of PromoCell Normal Human Cells

How should I handle cryopreserved cells upon receipt from PromoCell?

Abridged protocol: Take the cryopreserved cells out of the dry ice shipping container and place the cells immediately in a liquid nitrogen container. The interval between transfer from the shipping container to storage in the liquid nitrogen container should be as short as possible to prevent warming up. Do not store the cells at . 80°C as this causes irreversible damage to the cells.

For detailed information see: 1.2 Handling of PromoCell Normal Human Cells

How much experience do I need for starting with PromoCell normal human cell cultures?

You need experience in working under sterile conditions under a laminar flow hood, and it is of advantage having experience with other cell types.

If you are a beginner in cell culture or would like to establish a cell culture lab, we will assist you in working with PromoCell normal cells. You will learn how easy it is to work with PromoCell products. Please contact our Cell Culture Specialist.

Are PromoCell normal human cells from a single donor or are they pooled from several donors?

PromoCell normal human cells are derived from a single donor. Each shipment of normal human cell comes with a detailed certificate of analysis providing growth parameters and information about the donor's age, sex, and race.

What temperature and CO₂ content 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, unscrew the lid half a turn to ensure ventilation.

How often should I change the growth medium?

The medium should be changed every 2-3 days . most cell culture labs feed cells every Monday, Wednesday, and Friday.

Please note: After thawing PromoCell cryopreserved cells, the first medium change should be done 24 hours after seeding to prevent damage to the cells by residual cryomedium.

Do I need additional medium supplements to work with PromoCell cell growth media?

After adding SupplementMix or SupplementPack to the basal medium, you have the complete growth medium. No further addition is required.

How long can I use PromoCell supplemented growth media?

PromoCell supplemented growth media can be stored at 4°C for 6 weeks after adding SupplementMix or SupplementPack. Prior to use, warm up an aliquot of the growth medium at 37°C and keep the bottle with the remaining growth medium at 4°C. Please refer to "Should I warm up PromoCell cell culture medium prior to use?"

Should I warm up PromoCell cell culture medium prior to use?

PromoCell cell culture medium should have at least room temperature prior to use. If you want to warm up a portion of the medium at 37°C, do not apply uncontrolled heating. Please refer to "How long can I use PromoCell supplemented growth media?"

Should I warm up PromoCell trypsin prior to use?

The PromoCell DetachKit is delivered on dry ice. Thaw the solutions at room temperature and use them at room temperature. Do not use the solutions below or above room temperature.

Can I use my own medium or other commercially available media for culturing PromoCell normal human cells?

PromoCell normal human cells have been cultured and tested in our growth media and have adapted to this formula. Using other media for culturing PromoCell normal human cells may yield unsatisfactory growth results due to the adaptation process. We recommend the use of PromoCell media for culturing normal human cells.

Can I use my own trypsin or other commercially available trypsin solutions for subculturing PromoCell normal human cells?

The PromoCell DetachKit contains all components necessary for subculturing normal human cells. Each batch of trypsin solution has been tested with normal human cells. Using other trypsin solutions may cause irreversible damage to the cells. We recommend the use of PromoCell DetachKit for culturing normal human cells.

How many passages can I perform with PromoCell normal human cells?

PromoCell does not use the term "passage" for describing the growth potential because each customer uses a different split ratio. PromoCell guarantees 15 population doublings for normal human cells (unless otherwise indicated on the certificate of analysis) when using PromoCell cell culture media and the PromoCell DetachKit for the culturing procedure.

For detailed information see "1.7 Biological Growth Characteristics"

What is the recommended split ratio for PromoCell normal human cells?

PromoCell does not recommend a split ratio for normal human cells because the yield of cells after trypsinization varies. We recommend counting the cells after trypsinization and seeding the cells at 5,000 to 10,000 cells per cm² (find the recommended seeding density on the corresponding product information sheet).

For detailed information on how to count cells see "1.6 Counting the Cells".

Is it possible to subject PromoCell normal human cells to an experimental starvation phase?

Yes, it is possible to perform an experimental starvation phase with PromoCell normal human cells. Cells can be kept for some time in basal medium without supplements, but the cells have to be in a good condition. After this period, the experiment should be terminated because prolonged periods lead to cell death. It is advisable to test prior to the experiment how long cells will survive a starvation period; this depends on many factors.

Is it possible to carry out transient transfection with PromoCell normal human cells?

Yes, we have obtained good transfection results with many cell types (for example, human umbilical vein endothelial cells, normal human dermal fibroblasts, normal human osteoblasts).

For detailed information about transient transfection see "1.8 Transient Transfection of Normal Human Cells with Plasmid DNA (Lipofection Method)"