Thermo Scientific Centrifugal Small Class | How to Purify Exosomes by Ultracentrifugation

As the smallest unit of our lives, we have macro communication, and they naturally have micro communication. The "language" of this cell is the exosomal system.

The meaning of the exosomal system

Simply put, cells separate their own part of the membrane, wrap their "love words" (nucleic acids, proteins, etc.) into exosomes and release them, and finally integrate into other cells to communicate. For us humans, cracking the exosomal system is equivalent to cracking the password of cell communication, which is of great significance to the development of our life sciences.

Purification of exosomes by ultracentrifugation

The first step in studying exosomes must be to catch these little guys first. Although the exogenous body separation market is full of flowers. But these separation techniques are not satisfactory in terms of purity. Get the purest exosomes and uncover the story behind them. Ultracentrifugation is the best choice.

Then,

How to choose the right centrifuge

Perform centrifugation of exosomes?

Thermo Fisher is a world leader in scientific services, and its centrifuge products are known for their safety, reliability and meticulous convenience.

â–¶ Thermo Scientific Sorvall WX+ Series Ultracentrifuge

Excellent centrifugal performance: maximum centrifugal force of 802,000 xg, meeting the needs of exosomes purification experiments

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Recently, a new microfluidic device was developed by researchers at Massachusetts General Hospital (MGH). The device can use microfluidic technology to rapidly detect exocrine cells derived from cancer cells in the blood as a diagnostic tool for cancer.

Corresponding author Shannon Stott said their devices were able to select tumor-specific extracellular vesicles from billions of extracellular vesicles in the blood. This will provide new ways for future cancer diagnosis and monitoring. [1]

From this point of view, the application of exosomes can be said to be more and more extensive. This technology is also in line with today's hot liquid " liquid biopsy " thinking, the application prospects can be described as very bright. With regard to liquid biopsy, another simple and rude name is “blood cancer test”. Apparents who want to know about "liquid biopsy" can continue to follow our WeChat public account. We will describe in detail later.

So, how is ultracentrifugation applied in exosomes?

Purification of exosomes by ultracentrifugation

Reference experiment flow

1. First take the cell culture supernatant or body fluid as a sample, centrifuge at 300 xg for 10 min at low speed, and separate the cells; leave the supernatant at 2000 xg for 10 min to remove dead cells.

2. The remaining supernatant continues to increase the centrifugal force to 10000 xg for 30 min to remove cell debris, and the treated supernatant can be prepared for the ultracentrifugation step. Centrifuge for 70 min at a centrifugal force of 100,000 xg to obtain a crude exosome precipitate (containing a small amount of heteroprotein).

3. Resuspend the exosomal pellet with PBS, again at 100,000 xg, and centrifuge for 70 min to obtain a pure exosomes. [2]

How to choose the right centrifuge for centrifugation of exosomes?

â–¶Thermo ScientificTM SorvallTM WX+ Series Ultracentrifuge

Excellent centrifugal performance: maximum centrifugal force of 802,000 xg, meeting the needs of exosomes purification experiments

A wide selection of rotors for centrifugation from 1.5 mL centrifuge tubes to 250 mL centrifuge bottles for a wide range of applications in your laboratory for unparalleled convenience

Auto-LockTM III rotor self-locking system, 3 seconds to replace the rotor to achieve multi-purpose; Auto-ID instant rotor automatic identification

Third-party certified ClickSealTM bio-safe sealing technology to prevent aerosols from protecting the safety of laboratory personnel

Never fatigued FiberliteTM rotor, light weight, high hardness, corrosion resistance, 15 year warranty, no need to slow down for life

Recommended configuration:

*This program is for reference only and does not cover all rotors, baskets, adapters and centrifuge tubes. If you need to configure the solution in detail, please contact local sales.

Article Source:

[1] Engineered nanointerfaces for microfluidic isolation and molecular profiling of tumor-specific extracellular vesicles. Nat. Commun. 2018,9(1):175.

[2] Théry, Clotilde, et al. “Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological Fluids.” Current Protocols in Cell Biology, 2006, doi:10.1002/0471143030.cb0322s30.

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