What is RPMI culture medium?

RPMI 1640, also known as RPMI medium, is a growth medium used in cell culture. RPMI 1640 has traditionally been used for the serum-free expansion of human lymphoid cells. RPMI 1640 uses a bicarbonate buffering system and differs from most mammalian cell culture media in its typical pH 8 formulation.

How do you make an RPMI medium?

RPMI 1640 (11-100-1) – 1 liter

1. Prepare 900ml of distilled water in clean glass beaker.
2. Add the 1-liter powder to the water and stir gently.
3. Add 2 gram Sodium Bicarbonate (or 26.67 ml of 7.5% Sodium Bicarbonate Solution).
4. Adjust pH to 0.2-0.3 units below the required pH using 1N HCl or 1N NaOH.

Is RPMI a basal medium?

RPMI 1640 Medium uses a sodium bicarbonate buffer system (2.0 g/L), and therefore requires a 5–10% CO2 environment to maintain physiological pH. For in vitro diagnostic use. CAUTION: Not for human or animal therapeutic use. Uses other than the intended use may be a violation of local law.

Does RPMI contain FBS?

RPMI 1640 Medium contains no proteins, lipids, or growth factors. Therefore, RPMI 1640 Medium requires supplementation, commonly with 10% Fetal Bovine Serum (FBS).

Why is RPMI 1640 used?

RPMI-1640 medium has been used for the culture of human normal and neoplastic leukocytes. RPMI-1640, when properly supplemented, has demonstrated wide applicability for supporting growth of many types of cultured cells, including fresh human lymphocytes in the 72 hour phytohemaglutinin (PHA) stimulation assay.

What kind of cell types is RPMI medium used for?

RPMI is suitable for cells in suspension or monolayer and is widely used to grow a variety of mammalian cell types. These include primary human leukocytes, bone marrow cells, hybridomas, as well as common cell lines. RPMI typically requires supplementation with serum protein, such as FBS.

How do you make a medium RPMI 1640?

RPMI 1640 (media for YAC-1 cells)

1. adjust volume to 1 L.
2. adjust pH to 7.2.
3. sterile filter.
4. add 10 ml pen/strep (100x) to a final concentration of 100 U/ml penicillin, 100 µg/ml streptomycin.
5. add 10 % FBS.

How is medium prepared for cell culture?

Dehydrated media for broth cultures need to be prepared by dissolving in distilled water and adjusting the final pH prior to sterilization. Powdered media for agar cultures must be dissolved in distilled water, stirred, then boiled or autoclaved prior to pouring into sterile petri dishes using aseptic techniques.

Can I use Dmem instead of RPMI?

On a very basic level in our laboratory you would use DMEM for an adherent cell line and you would use RPMI for suspension cell lines. For example some cell lines grow and flourish better in the presence of HEPEs. Also you need to think about your down the line uses for the cells.

How do you make a fetal bovine serum?

The first stage of the production process for FBS is the harvesting of blood from the bovine fetus after the fetus is removed from the slaughtered cow. The blood is collected aseptically into a sterile container or blood bag and then allowed to clot.

What is in Hepes buffer?

Gibco HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid ) is a zwitterionic organic chemical buffering agent and is categorized as a “Good” buffer which derives from a set of buffers described by Dr. Norman Good and his colleagues in 1966 (Good et.

How much RPMI is strep pen?

You can take out 5.56 mL RPMI from your original 500 mL stock and add there 5.56 mL 100% Pens-Strep, then add there 56 mL FBS. Mix well. Hope it helps.

Is there a complete formulation of RPMI 1640?

The complete formulation is available. RPMI 1640 Medium is unique from other media because it contains the reducing agent glutathione and high concentrations of vitamins. RPMI 1640 Medium contains biotin, vitamin B 12, and PABA, which are not found in Eagle’s Minimal Essential Medium or Dulbecco’s Modified Eagle Medium.

What is the difference between RPMI 1640 medium and Eagle medium?

RPMI 1640 Medium is unique from other media because it contains the reducing agent glutathione and high concentrations of vitamins. RPMI 1640 Medium contains biotin, vitamin B 12, and PABA, which are not found in Eagle’s Minimal Essential Medium or Dulbecco’s Modified Eagle Medium. We offer a variety of RPMI 1640 Medium…

What are the ingredients in RPMI?

In addition, the vitamins inositol and choline are present in very high concentrations. This RPMI formulation has been modified by ATCC to contain high glucose, low sodium bicarbonate, sodium pyruvate, HEPES, and low L-glutamine.

Does 1640 medium contain biotin?

RPMI 1640 Medium contains biotin, vitamin B 12, and PABA, which are not found in Eagle’s Minimal Essential Medium or Dulbecco’s Modified Eagle Medium. In addition, the vitamins inositol and choline are present in very high concentrations.