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Exosomes are a type of vesicles secreted by cells to the outside of the cell, with a size of 30-150nm, a double-layer membrane structure and a saucer-like shape, and rich contents (including nucleic acids, proteins, lipids, etc.
) Molecular transfer between cells
.
Exosomes are widely found in cell culture supernatants and various body fluids, and the research on exosomes has become a hot research topic in recent years
.
) Molecular transfer between cells
.
Exosomes are widely found in cell culture supernatants and various body fluids, and the research on exosomes has become a hot research topic in recent years
.
The main principle of cellular exosomes is to separate exosomes based on differences in density, particle size, sedimentation coefficient and affinity.
Extraction methods include traditional differential centrifugation, density gradient centrifugation, and later developed ultrafiltration and polymer precipitation.
method, immune separation, isolation screening method, size exclusion chromatography,
etc.
Each of these methods has pros and cons
.
Extraction methods include traditional differential centrifugation, density gradient centrifugation, and later developed ultrafiltration and polymer precipitation.
method, immune separation, isolation screening method, size exclusion chromatography,
etc.
Each of these methods has pros and cons
.
Frequently asked questions about cell exosome research:
1.
How to avoid serum-derived exosome contamination when culturing cells?
How to avoid serum-derived exosome contamination when culturing cells?
Cell culture is inseparable from serum, and serum generally contains exosomes.
In order to avoid the interference of serum exosomes, it is recommended to use ultracentrifugation to make exosome-free serum or use commercialized exosome-free serum for cell culture.
cultivate
.
In order to avoid the interference of serum exosomes, it is recommended to use ultracentrifugation to make exosome-free serum or use commercialized exosome-free serum for cell culture.
cultivate
.
2.
Will dead cells during cell culture affect the extraction of exosomes?
Will dead cells during cell culture affect the extraction of exosomes?
Yes
.
The process of apoptosis will release a large number of vesicles, which will contaminate the exosomes produced by living cells during the extraction and purification of exosomes
.
Therefore, when harvesting cells, the proportion of dead cells should not exceed 5%
.
.
The process of apoptosis will release a large number of vesicles, which will contaminate the exosomes produced by living cells during the extraction and purification of exosomes
.
Therefore, when harvesting cells, the proportion of dead cells should not exceed 5%
.
3.
How much volume is needed to isolate cell supernatant exosomes?
How much volume is needed to isolate cell supernatant exosomes?
Different cell lines require different amounts
.
For cell lines with high release, a few milliliters of cell supernatant are sufficient; for cell lines with low release, hundreds of milliliters may be required, and preliminary experiments are required
.
.
For cell lines with high release, a few milliliters of cell supernatant are sufficient; for cell lines with low release, hundreds of milliliters may be required, and preliminary experiments are required
.
The study of cellular exosomes is usually closely related to the study of high-throughput sequencing, and can be roughly divided into three categories:
1.
Expression profile: It is characterized by being short, flat and fast, usually with sequencing data as the main content, with an impact factor of 3-5;
Expression profile: It is characterized by being short, flat and fast, usually with sequencing data as the main content, with an impact factor of 3-5;
2.
Molecular markers: It is characterized by adding a large number of samples to verify the expression profile, establishing ROC and KM curves, and analyzing the correlation between molecules and clinical diseases.
The article affects the shadow 3-10;
Molecular markers: It is characterized by adding a large number of samples to verify the expression profile, establishing ROC and KM curves, and analyzing the correlation between molecules and clinical diseases.
The article affects the shadow 3-10;
3.
Molecular mechanism research: To achieve in-depth research on cell function and mechanism, the workload is large and the impact factor is also high
.
Molecular mechanism research: To achieve in-depth research on cell function and mechanism, the workload is large and the impact factor is also high
.
