Dialyzed vs Charcoal Stripped FBS
The blood component called serum is what remains of the cells after coagulation and elimination. In addition to serum proteins, it also contains hormones, amino acids, and growth factors. The serum is one of the most crucial supplements for promoting cell development and proliferation in vitro cell culture because of its comprehensive mixture. Fetal bovine serum is very interesting (FBS).
It is particularly high in growth factors and low in antibodies, which could affect the effectiveness of cell culture.
What is FBS, or charcoal-stripped?
Activated charcoal is used to completely remove free, low molecular weight hormones and proteins from charcoal-stripped plasma and serum. This method does not guarantee the removal of hormones linked to proteins.
Usage of charcoal-stripped?
Studies on hormone-responsive malignancies typically use a charcoal-stripped fetal bovine serum (CS-FBS) to establish hormone-free cell culture conditions. Although the underlying mechanisms are still unknown, CS-FBS may have an impact on the development of cancer cells. When examining androgen responsiveness and androgen metabolism in cultured prostate cancer (CaP) cells, charcoal-stripped fetal bovine serum (CS-FBS) is frequently utilised.
Without eliminating nutrients, charcoal-stripped FBS eliminates nonpolar molecules such as viruses, certain growth factors, hormones, and cytokines (salts, glucose, amino acids). Highly effective for studies on hormones, cancer, cell signalling, medication response, etc.
Benefits of charcoal-stripped
The process of filtering fetal bovine serum (FBS) through an activated carbon-absorbent filter reduces the quantities of steroid hormones such as estradiol, progesterone, cortisol, testosterone, T3, and T4 without losing other serum components in an unintended way. FBS Charcoal Stripped helps research processes affected by steroid hormones (oestrogen stimulation, obesity process), as well as other research areas that profit from low hormone levels, like particular kinds of viral infections.
Widely utilised in eukaryotic cell culture, fetal calf or bovine serum is typically added to growth media as a supplement. A good cell culture depends on the nutritional and growth-promoting components found in fetal bovine serum. Additionally, the fetal bovine serum has a variety of tiny molecules, including hormones, lipids, carbohydrates, and amino acids.
The blood component obtained following coagulation and the removal of cellular components is called fetal bovine serum (FBS). FBS is one of the most crucial supplements for promoting cell growth and proliferation in vitro cell culture because of its distinctive and thorough mix.
Charcoal-Stripped Serum Preparation
- Incubate at 4 °C all night. Dextran-coated charcoal or Activated Charcoal Norit in a solution of 0.25 M sucrose, 1.5 mM MgCl2, and 10 mM HEPES at a pH of 7.4 and Dextran T-70 at a final concentration of 0.0025 per cent.
- Use an amount of dextran-coated charcoal (DCC) from step 1 that is equal to the volume of the serum that needs to be stripped. To pelletize the charcoal, centrifuge it at 500 x g for 10 minutes.
- Substitute the same volume of FBS for the supernatant. Keep in mind that various FBS lots, even those from the same source, may have various growth characteristics and must be verified using the tissue culture procedure used in your lab.
- After completely mixing the charcoal with the serum using a vortex, the tube should be incubated for 12 hours at 4 °C (for DCC-stripped serum) or twice for 45 minutes at 56 °C (for heat-inactivated, DCC-stripped serum, or HIDCC serum).
- The complement-deficient serum is also made via heat-inactivation.
Dialyzed FBS: What is it?
Benefits of Dialyzed FBS
- using a 12–14 kDa cutoff membrane for dialysis
- using glucose monitoring to control a process
- to avoid peptide precipitation and inactivation in the FBS during dialysis
- high cell growth efficiency and low endotoxin
- Mexican/Central American FBS with USDA approval
- produced in a facility with an ISO 9001:2015 certification
- was distributed under the Atlanta Biologicals name
Why Use Heat Inactivated, Dialyzed Fetal Bovine Serum?
A 12,000 to 14,000 Dalton cutoff membrane is used to dialyze pre-selected fetal bovine serum, also known as fetal calf serum (FCS), versus physiological saline. Throughout the operation, the glucose concentration is carefully monitored to keep the process under control. Serum peptide precipitation and inactivation are not prevented by exhaustive dialysis.
Storage and Handling
FBS - Dialyzed 12-14 kD is provided in sterile, gamma-irradiated PETG or PETE bottles. We advise keeping the serum refrigerated between -5 and -20 degrees Celsius. The serum should not undergo repeated freeze-thaw cycles since this could cause the product's quality to decline. We advise using one of our smaller container sizes or separating the serum into smaller aliquots suitable for single-use if occasional consumption of the product is anticipated. When handling the serum, always use aseptic procedures, and a liquot into sterile containers.
Precaution
Safe laboratory practices and personal protective equipment should be used when handling biohazardous materials like human cells.
Wearing protective gear is advised.
Limitations
Not to be used in diagnostic procedures; only for laboratory research use.
- It is unknown whether this product is safe and effective for diagnostic or other clinical applications.
- Because tissue/cells from various donors or sources differ, results may vary.
- Libny Auguste