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Exosome Isolation from Cell Culture Media

Exosome isolation from cell culture is a crucial step when trying to understand how intercellular communication is happening and the influence between the two. In this presentation, we look at the advantages and disadvantages of common exosome isolation methods from cell culture for different research purposes. To compare the isolated exosome preparation with a sample containing exosomes of interest, you can contact us at 919654321400 to procure a pure sample of exosomes isolated from human and preclinical animal cells.<br><br><br>For more Info Visit: www.kosheeka.com

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Exosome Isolation from Cell Culture Media

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  1. EXOSOME ISOLATION FROM CELL CULTURE MEDIA Isolating the Nano-Wonder from Cell Culture

  2. EXOSOMES: THE EXTRACELLULAR VESICLE • Exosomes are the smallest extracellular vesicle, with their size ranging from 30 nm to 150 nm. • These are important communication mediators for intercellular communication. • Purity of the exosome sample is necessary to elucidate its function and role. • Exosomes also carry specific markers, making them necessary for specific research studies and targeted therapies. www.kosheeka.com

  3. HOW DO WE ISOLATE EXOSOMES? • There are several methods to isolate exosomes, including • Precipitation • Microfluidics • Nanolithography • Electro-deposition • Immunomagnetic beads • Covalent chemistry • Ultra-centrifugation • Size-exclusion Chromatogaphy Source: PMID: 36714629 www.kosheeka.com

  4. COMMON METHOD OF ISOLATION #1 PRECIPITATION RATIONAL Samples with exosomes are incubated with a solution that changes their solubility or sedimentation rate. ADVANTAGES DISADVANTAGES • High Yield of extracellular vesicles • High sample process rate • Simple procedure • Do not reuquire specialized equipment • Maintain vescile,s morphology and integrity • Difficult toisolate specific type of exosomes • Can carry protein aggregares, lipoproteins, and viruses • Can be contaminated with precipitatiing polymer or chemical www.kosheeka.com

  5. COMMON METHOD OF ISOLATION #2 ULTRAFILTRATION RATIONAL The ultrafiltration process allows the sample solution to pass over a semi-permeable membrane to allow exosomes of a specific size to pass through while retaining others in the sample solution. Tangential Flow Ultrafiltration ADVANTAGES DISADVANTAGES • SImplified protocol • Yields pure exosome samples • Multiple sample processing • Concentrate exosome isolation from dilute samples • Reduced filter clogging • Some loss of sample in filtration • Clogging of the filter • Distortion in the exosome morphology • Contamination with protein aggregates Source: PMID: 32206116 www.kosheeka.com

  6. COMMON METHOD OF ISOLATION #3 ULTRACENTRIFUGATION RATIONAL Samples undergo several rounds of centrifugation (100,000–200,000 g) to remove cellular debris from the sample, followed by diffential centrifugation to yield pure exosomes. ADVANTAGES DISADVANTAGES • Isolation from large volumes • No added chemicals • Require an ultracentrifugation mean • Low yield • Damage to exosomes at high speeds • Sample viscosity impacts sedimentaton • Cannot scale up to multiple samples Source: PMID: 35087806 www.kosheeka.com

  7. COMMON METHOD OF ISOLATION #4 SIZE-EXCLUSION CHROMATOGRAPHY RATIONAL The exosomes are isolated based on their size as the sample passes through a porous column (pore size 40–80 nm), and the exosomes are collected as eluation. ADVANTAGES DISADVANTAGES • Highly reproducible • Preserves the exosomes structure • Easily scalable and can work with multiple samples • Efficient in segrating exosomes based on sizes • Easy to customize to get exosomes of desired size • Restrict sample volumes • Low resolution of similar sized molecules • One type of exosomesi processed at a given time www.kosheeka.com

  8. CONCLUSION: • Exosomes are crucial for intercellular communication. • Exosome isolation is challenging due to size and heterogeneity. • Methods for exosome isolation include: • Precipitation: simple, high yield, low purity • Ultrafiltration: efficient, prone to clogging • Ultracentrifugation: gold standard, laborious, potentially damaging • Size-exclusion chromatography: high purity, limited sample volume • A combination of methods may be necessary. www.kosheeka.com

  9. THANK YOU FOR ATTENTION www.kosheeka.com info@kosheeka.com +91-96543-21400

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