Exosome Isolation from Cell Culture Media
                     EXOSOME
ISOLATION FROM CELL 
CULTURE MEDIA
Isolating the Nano-Wonder from Cell 
Culture
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.
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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: 
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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 • Difficult toisolate specific type of 
• High sample process rate exosomes
• Simple procedure • Can carry protein aggregares, 
• Do not reuquire specialized equipment lipoproteins, and viruses
• Maintain vescile,s morphology and • Can be contaminated with 
integrity precipitatiing polymer or chemical
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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 • Some loss of sample in filtration
• Yields pure exosome samples • Clogging of the filter
• Multiple sample processing • Distortion in the exosome 
• Concentrate exosome isolation from morphology
dilute samples • Contamination with protein 
• Reduced filter clogging aggregates
Source: PMID: 
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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 • Require an ultracentrifugation mean
• No added chemicals • Low yield
• Damage to exosomes at high speeds
• Sample viscosity impacts 
sedimentaton
• Cannot scale up to multiple samples
Source: PMID: 
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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 • Restrict sample volumes
• Preserves the exosomes structure • Low resolution of similar sized 
• Easily scalable and can work with molecules
multiple samples • One type of exosomesi processed at 
• Efficient in segrating exosomes based a given time
on sizes
• Easy to customize to get exosomes of 
desired size
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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.
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THANK YOU
FOR ATTENTION
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[email protected]
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