Discover How Exosome Depletion Creates Optimized FBS
Fetal bovine serum (FBS) is an essential supplement used in cell culture media to provide nutrients and growth factors for optimal cell health and performance. However, standard FBS contains bovine-derived exosomes that can interfere with experiments examining human cell-secreted exosomes.
This article will examine what exosome-depleted FBS is, why it’s used, where to source it, how it’s prepared, and how it differs from regular FBS.
What is Exosome-Depleted FBS?
Exosome-depleted FBS refers to fetal bovine serum that has been processed to remove or deplete extracellular vesicle particles known as exosomes.
Exosomes are small lipid bilayer vesicles (30-150 nm) secreted by cells and found in bodily fluids. They act as transport vehicles to transfer molecules between cells and play diverse signaling roles.
FBS naturally contains bovine-derived exosomal vesicles. To create an optimized cell culture supplement for exosome studies, methods like ultracentrifugation or filtration are used to remove or significantly reduce exosomes from standard FBS.
The resulting exosome-depleted FBS aims to provide a clean slate, removing any confounding bovine exosomal contaminants. This allows researchers to isolate and examine exosomes secreted specifically by human cell cultures, without interference from residual bovine exosomes in the FBS supplement.
What is 10% Exosome-Depleted FBS Used For?
Like regular FBS, exosome-depleted FBS is added as a supplement when formulating cell culture media. Typical concentrations are around 5-10%. At this level it provides necessary nutrients, growth factors, and proteins to nourish cells and maintain viability.
Simultaneously, it eliminates bovine exosomes that could interfere with experiments focused on isolating and analyzing exosomes secreted by the cells in culture. Using exosome-depleted FBS as the media supplement allows researchers to study the cell’s secreted exosomes with confidence.
The Purpose of Exosome-Depleted Medium
The goal of formulating cell culture media with exosome-depleted FBS is to remove bovine-derived exosomes while retaining serum components that support cell growth. Excluding exogenous FBS-sourced exosomes aims to:
- Prevent contamination of cell culture exosome isolates
- Enable detection of low abundance human exosomes
- Standardize basal exosome levels across experiments
- Eliminate confounding factors from bovine exosomes
This optimized cell culture medium provides a cleaner background to elucidate exosomes secreted specifically by the cells under study. However, batch variability in exosome depletion efficiency should be assessed when comparing results across studies.
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Where Can I Source Exosome-Depleted FBS?
Several leading life science companies offer ready-made exosome-depleted FBS products manufactured under controlled conditions:
| Brand | Product | Description |
| Gibco | Exosome-Depleted FBS | Performance tested and optimized for exosome isolation and characterization studies. |
| System Biosciences | Exo-FBS | Specifically designed for exosome secretion experiments. |
| Thermo Fisher | Exosome-Depleted FBS | Maintains high exosome depletion (>90%) with consistent cell culture performance. |
| Norgen Biotek | FBS Exosome Depletion Kit | Provides components to remove exosomes from standard FBS in the lab. |
| South America Fetal Bovine Serum | Exosome-Depleted FBS Variant | Tailored for cell-derived exosome research applications. |
Compare options to find the right exosome-depleted FBS for your specific protocol and study aims. Batch-to-batch consistency is a key factor when selecting a product.
Exosome-Depleted FBS Preparation
Exosome-depleted FBS can be prepared in the lab or purchased as a commercially available product. If purchasing pre-made exosome-depleted FBS, be sure to check the manufacturer’s technical documentation to understand their proprietary methods for exosome removal.
Common methods for preparing exosome-depleted FBS in the lab include:
Ultracentrifugation
This involves centrifuging FBS at very high speeds of 100,000 x g or greater for 18 hours or longer. The high g-forces pellet exosomes, allowing removal of the clarified supernatant which is depleted of exosomes.
A major advantage of this method is very high exosome removal efficiency. However, it requires an expensive ultracentrifuge and lengthy spin times.
Filtration
FBS can be passed through 0.1 μm membrane filters to physically exclude exosomes based on size. This is rapid and simple, but may not remove all exosomes if smaller vesicles are present. Success depends on the pore size chosen.
Commercial Kits
Companies like Norgen Biotek provide kits containing spin columns and protocols to easily deplete exosomes from standard FBS in the lab. This provides standardized materials and methods for consistent results. However, kit performance should still be validated.
Dialysis
Dialyzing FBS overnight against an appropriate buffer and membrane molecular weight cutoff can also gently remove exosomes. This avoids the high g-forces of centrifugation that could damage proteins. However, dialysis may be less efficient than other methods.
The optimal method for a given application depends on balancing exosome removal efficiency with ease of use and cost. Rigorously testing the depletion efficiency and culture performance of any exosome-depleted FBS prepared in the lab is critical to ensuring suitability as a cell culture supplement.
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How Exosome-Depleted Serum Differ from Regular Serum
Exosome-depleted serum differs from regular serum in several ways. Studies have found significant differences in the composition of exosome fractions compared to exosome-depleted fractions of fetal bovine serum (FBS). Some of the key differences include:
- Exosome fractions contain extracellular vesicles and associating proteins/RNA left behind after depletion.
- Multiple microRNA levels decreased by 10 to 100-fold after FBS exosome depletion.
- Growth factors like FGF were reduced in exosome-depleted FBS.
- Overall profiles of proteins, lipids, and RNA content change with exosome removal.
Exosome depletion leaves behind the soluble serum components, but the extracellular vesicle population and its molecular cargo are specifically excluded. This alters the functional properties of depleted serum relative to complete FBS.
Exosome-depleted FBS has been used in various studies to eliminate the risks associated with FBS-induced cell migration and to streamline cell culture workflows for exosome research.
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