CN110747158A - Cell supernatant exosome extraction process based on precipitation reagent method - Google Patents
Cell supernatant exosome extraction process based on precipitation reagent method Download PDFInfo
- Publication number
- CN110747158A CN110747158A CN201911114046.5A CN201911114046A CN110747158A CN 110747158 A CN110747158 A CN 110747158A CN 201911114046 A CN201911114046 A CN 201911114046A CN 110747158 A CN110747158 A CN 110747158A
- Authority
- CN
- China
- Prior art keywords
- supernatant
- exosomes
- under
- aseptic condition
- cell supernatant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Abstract
The invention discloses a cell supernatant exosome extraction process based on a precipitation reagent method, belonging to the field of exosome extraction. The invention relates to a method for combining multi-time centrifugation and a precipitating agent, which comprises the steps of firstly removing cells and cell fragments through centrifugation, removing impurities of macromolecules through a filter membrane, improving the purity of exosomes in supernatant, combining PEG (polyethylene glycol) with a lipid molecule layer of the exosomes, changing the solubility or dispersibility of the exosomes, precipitating the exosomes, centrifuging to obtain primary exosomes, mixing the primary exosomes with a mixed solution of ethanol and ether, removing a small amount of low-abundance proteins in the primary exosomes to obtain high-purity exosomes, combining a streptavidin precipitating agent with the resuspended high-purity exosomes, combining a specific magnetic probe in the streptavidin precipitating agent with the exosomes, centrifuging to obtain precipitates, further removing the low-abundance proteins, and further improving the purity of the exosomes.
Description
Technical Field
The invention relates to the field of exosome extraction, in particular to a cell supernatant exosome extraction process based on a precipitation reagent method.
Background
The exosome is used as an important component of paracrine, and is a lipid bilayer membrane vesicle-like structure corpuscle secreted by various living cells, the size of which is about 30-100 nm and the density of which is 1.13-1.19 g/ml, and the lipid bilayer membrane vesicle-like structure corpuscle is fused with a plasma membrane and then released into an extracellular environment in a exocytosis mode. When the exosome is contacted with a target cell, the exosome and its carried biomolecules (e.g., functional lipids, proteins, molecules such as messenger rnas (mrnas), microRNAs, etc.) are taken up and acted upon in an endocytic form. A great deal of research also finds that the stem cells can play the roles of tissue repair, immunosuppression and immune function regulation similar to cells through self exosomes.
At present, differential centrifugation and precipitation kit methods are mainly adopted for exosome extraction. The differential centrifugation method mainly realizes the extraction of exosome by alternately carrying out low-speed centrifugation and high-speed centrifugation, although the method has simple operation and more and popular vesicles, the process is time-consuming, the recovery rate is unstable, the purity is questioned, and in addition, the repeated centrifugation operation can possibly damage the vesicles, thereby reducing the quality of the vesicles; the precipitation kit method mainly combines a precipitator with hydrophobic protein and lipid molecules to form coprecipitation, so as to realize the extraction of exosome, and although the method is simple in operation and low in cost, the specificity is not high, and not only can high-abundance proteins be precipitated, but also part of low-abundance proteins can be precipitated together.
Based on the above, the invention designs a cell supernatant exosome extraction process based on a precipitation reagent method, so as to solve the above problems.
Disclosure of Invention
The invention aims to provide a cell supernatant exosome extraction process based on a precipitation reagent method, so as to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: a cell supernatant exosome extraction process based on a precipitation reagent method comprises the following steps:
s1, putting cells needing to extract exosomes into a culture medium, and carrying out 3-6% CO treatment at 36-38 DEG C2Culturing the seeds by attaching to the wall for 70-72 d under the aseptic condition;
s2, under the aseptic condition, taking 0.5-1.0 mL of supernatant of the S1 culture medium, placing the supernatant into a centrifuge at the temperature of 2-6 ℃ and 1500-2000 Xg for 10-20 min, discarding the bottom precipitate after centrifugation, and obtaining cell supernatant;
s3, under the aseptic condition, the cell supernatant in the S2 passes through a 0.22 mu m filter to obtain a filtered supernatant;
s4, shaking the filtered supernatant in the S3 and 10-40% of PEG solution uniformly according to the volume ratio of 1: 4-8 under an aseptic condition, standing for 10-12 h at the temperature of 2-6 ℃, centrifuging for 20-30 min at 2500-3000 Xg, discarding the centrifuged supernatant, and obtaining a bottom precipitate by a back-off absorption paper method;
s5, under an aseptic condition, adding 5-7.5 mL of PBS (pH7.2-7.4) into the bottom precipitate in the S3, carrying out heavy suspension, adding 0.5-1.0 mL of 6.0-10.0 mg/mL of streptavidin precipitator, uniformly shaking, and standing for 20-30 min at 2-8 ℃;
s6, centrifuging the liquid obtained after S5 is stood at 2500-3000 Xg for 20-30 min at the temperature of 2-8 ℃ under the aseptic condition, discarding the supernatant, and removing magnetic beads by using a magnetic frame to obtain a precipitate, namely the required exosome;
s7, adding 0.5-1.0 ml PBS solution (pH7.2-7.4) into the exosome in the S6, carrying out heavy suspension, and then refrigerating at-70 to-80 ℃.
Furthermore, the culture medium in the S1 is one of RPMI1640 culture medium and DMEM culture medium.
Furthermore, the PEG in S4 is selected from one of PEG6000 and PEG 8000.
Furthermore, the concentration of PEG in the S4 is 10-40%.
Further, the inverted absorbent paper method in S4 includes the following steps:
A. reversely buckling the centrifugal tube which is centrifuged and then supernatant fluid is removed on absorbent paper, and air-drying for 5-10 min under the aseptic condition of 2-8 ℃;
B. adding 0.5-1.0 mL of ethanol and ether mixed solution (1:1, V/V) into the air-dried bottom sediment, shaking, centrifuging at 2-6 ℃ and 1500-2000 Xg for 10-20 min, discarding the centrifuged supernatant, reversely covering the centrifuged centrifuge tube on absorbent paper, and air-drying at 2-8 ℃ for 5-10 min under an aseptic condition to obtain the bottom sediment.
By adopting the technical scheme, the invention has the beneficial effects that: the invention relates to a method for combining multi-time centrifugation and a precipitating agent, which comprises the steps of firstly removing cells and cell fragments through centrifugation, removing impurities of macromolecules through a filter membrane, improving the purity of exosomes in supernatant, combining PEG (polyethylene glycol) with a lipid molecule layer of the exosomes, changing the solubility or dispersibility of the exosomes, precipitating the exosomes, centrifuging to obtain primary exosomes, mixing the primary exosomes with a mixed solution of ethanol and ether, removing a small amount of low-abundance proteins in the primary exosomes to obtain high-purity exosomes, combining streptavidin precipitating agent with resuspended high-purity exosomes, combining a specific magnetic probe in the streptavidin precipitating agent with the exosomes, centrifuging to obtain precipitates, further removing the low-abundance proteins, and further improving the purity of the exosomes.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without any inventive step, are within the scope of the present invention.
Example 1
A cell supernatant exosome extraction process based on a precipitation reagent method comprises the following steps:
s1, putting the cells to be extracted into culture medium, and culturing at 36 deg.C with 5% CO2Culturing the seeds on the wall for 70d under the aseptic condition;
s2, under the aseptic condition, taking 0.5mL of supernatant of the S1 culture medium, placing the supernatant at 4 ℃, centrifuging at 2000 Xg for 10min, discarding the bottom precipitate after centrifugation, and obtaining cell supernatant;
s3, under the aseptic condition, the cell supernatant in the S2 passes through a 0.22 mu m filter to obtain a filtered supernatant;
s4, shaking the filtered supernatant in S3 and 20% PEG solution uniformly according to the volume ratio of 1:4 under the aseptic condition, standing at 2 ℃ for 12h, centrifuging at 2500 Xg for 30min, discarding the centrifuged supernatant, and obtaining a bottom precipitate by an inverted absorption paper method;
s5, adding 7.5mL of PBS (pH7.3) into the bottom precipitate in the S3 under aseptic conditions, re-suspending, adding 0.8mL of 6.0mg/mL streptavidin precipitator, uniformly shaking, and standing at 2 ℃ for 27 min;
s6, centrifuging the liquid obtained after S5 is stood at 2 ℃ under aseptic conditions for 25min at 3000 Xg, discarding the supernatant, and removing the magnetic beads by using a magnetic frame to obtain a precipitate, namely the required exosome;
s7, 1.0ml PBS solution (pH7.2) was added to the exosomes in S6, and after resuspension, it was refrigerated at-70 ℃.
The culture medium in the S1 is RPMI1640 culture medium.
PEG6000 is selected as PEG in S4.
The concentration of PEG in S4 is 30%.
The operation steps of the inverted absorbent paper method in S4 are as follows:
A. reversely buckling the centrifugal tube which is centrifuged and then removed with supernatant on absorbent paper, and air-drying for 10min at 6 ℃ under the aseptic condition;
B. adding 0.5mL of mixed solution (1:1, V/V) of ethanol and ether into the air-dried bottom precipitate, shaking, centrifuging at 4 ℃ and 1800 Xg for 15min, discarding the centrifuged supernatant, reversely covering the centrifuged centrifuge tube on absorbent paper, and air-drying at 4 ℃ for 5min under aseptic condition to obtain the bottom precipitate.
Example 2
A cell supernatant exosome extraction process based on a precipitation reagent method comprises the following steps:
s1, putting the cells to be extracted into culture medium, and heating at 37 deg.C with 4% CO2Culturing the cells in a wall-mounted manner for 72d under the aseptic condition;
s2, under the aseptic condition, taking 0.6mL of supernatant of the S1 culture medium, placing the supernatant at 6 ℃ and centrifuging the supernatant at 1700 Xg for 18min, discarding bottom sediment after centrifugation, and obtaining cell supernatant;
s3, under the aseptic condition, the cell supernatant in the S2 passes through a 0.22 mu m filter to obtain a filtered supernatant;
s4, shaking the filtered supernatant in S3 and 40% PEG solution uniformly according to the volume ratio of 1:6 under the aseptic condition, standing for 10 hours at 4 ℃, centrifuging for 26 minutes at 3000 Xg, discarding the centrifuged supernatant, and obtaining a bottom precipitate by an inverted absorption paper method;
s5, adding 5mL of PBS (pH7.4) into the bottom precipitate in the S3 under aseptic conditions, re-suspending, adding 0.6mL of streptavidin precipitator of 7.0mg/mL, uniformly shaking, and standing at 4 ℃ for 24 min;
s6, centrifuging the liquid obtained after S5 is stood at 2500 Xg for 20min at 4 ℃ under aseptic conditions, discarding the supernatant, and removing the magnetic beads by using a magnetic frame to obtain a precipitate, namely the required exosome;
s7, 0.7ml PBS solution (pH7.2) was added to the exosomes in S6, and after resuspension, the exosomes were refrigerated at-70 ℃.
The culture medium in the S1 is RPMI1640 culture medium.
PEG6000 is selected as PEG in S4.
The concentration of PEG in S4 is 20%.
The operation steps of the inverted absorbent paper method in S4 are as follows:
A. reversely buckling the centrifugal tube which is centrifuged and then supernatant fluid is removed on absorbent paper, and air-drying for 8min at 2 ℃ under the aseptic condition;
B. adding 0.8mL of mixed solution (1:1, V/V) of ethanol and ether into the air-dried bottom precipitate, shaking, centrifuging at 4 ℃ and 2000 Xg for 10min, discarding the centrifuged supernatant, reversely covering the centrifuged centrifuge tube on absorbent paper, and air-drying at 6 ℃ for 8min under aseptic condition to obtain the bottom precipitate.
Example 3
A cell supernatant exosome extraction process based on a precipitation reagent method comprises the following steps:
s1, putting the cells to be extracted into culture medium, and heating at 37 deg.C with 6% CO2Culturing the seeds on the wall for 70d under the aseptic condition;
s2, under the aseptic condition, taking 0.8mL of supernatant of the S1 culture medium, placing the supernatant at 2 ℃ and centrifuging at 1900 Xg for 14min, discarding bottom sediment after centrifugation, and obtaining cell supernatant;
s3, under the aseptic condition, the cell supernatant in the S2 passes through a 0.22 mu m filter to obtain a filtered supernatant;
s4, shaking the filtered supernatant in S3 and 30% PEG solution uniformly according to the volume ratio of 1:7 under aseptic condition, standing at 4 ℃ for 11h, centrifuging at 2800 Xg for 24min, discarding the centrifuged supernatant, and obtaining a bottom precipitate by an inverted absorption paper method;
s5, adding 6.0mL of PBS (pH7.2) into the bottom precipitate in the S3 under aseptic conditions, re-suspending, adding 0.5mL of 9.0mg/mL streptavidin precipitator, uniformly shaking, and standing at 6 ℃ for 30 min;
s6, centrifuging the liquid obtained after S5 is stood at 2700 Xg for 25min at 6 ℃ under the aseptic condition, discarding the supernatant, and removing the magnetic beads by using a magnetic frame to obtain a precipitate, namely the required exosome;
s7, 0.5PBS (pH7.4) was added to the exosomes in S6, and after resuspension, the exosomes were refrigerated at-80 ℃.
The culture medium in the S1 is DMEM culture medium.
PEG8000 is selected as PEG in S4.
The concentration of PEG in S4 is 10%.
The operation steps of the inverted absorbent paper method in S4 are as follows:
A. reversely buckling the centrifugal tube which is centrifuged and then supernatant fluid is removed on absorbent paper, and air-drying for 5min at 4 ℃ under the aseptic condition;
B. adding 0.8mL of mixed solution (1:1, V/V) of ethanol and ether into the air-dried bottom precipitate, shaking, centrifuging at 6 ℃ and 1800 Xg for 15min, discarding the centrifuged supernatant, reversely covering the centrifuged centrifuge tube on absorbent paper, and air-drying at 2 ℃ for 8min under aseptic condition to obtain the bottom precipitate.
Example 4
A cell supernatant exosome extraction process based on a precipitation reagent method comprises the following steps:
s1, putting the cells to be extracted into culture medium, and heating at 38 deg.C with 3% CO2Culturing the cells in a wall-mounted manner for 72d under the aseptic condition;
s2, under the aseptic condition, taking 1.0mL of supernatant of the S1 culture medium, placing the supernatant at 4 ℃, centrifuging at 1500 Xg for 20min, discarding the bottom precipitate after centrifugation, and obtaining cell supernatant;
s3, under the aseptic condition, the cell supernatant in the S2 passes through a 0.22 mu m filter to obtain a filtered supernatant;
s4, shaking the filtered supernatant in S3 and 10% PEG solution uniformly according to the volume ratio of 1:8 under aseptic condition, standing at 6 ℃ for 12h, centrifuging at 2600 Xg for 20min, discarding the centrifuged supernatant, and obtaining a bottom precipitate by an inverted absorption paper method;
s5, adding 6.5mL of PBS (pH7.3) into the bottom precipitate in the S3 under aseptic conditions, re-suspending, adding 1.0mL of 10.0mg/mL streptavidin precipitator, uniformly shaking, and standing at 8 ℃ for 20 min;
s6, centrifuging the liquid obtained after S5 is stood at 2900 Xg for 30min at 8 ℃ under the aseptic condition, discarding the supernatant, and removing the magnetic beads by using a magnetic frame to obtain a precipitate, namely the required exosome;
s7, 0.8ml PBS solution (pH7.4) was added to the exosomes in S6, and after resuspension, the exosomes were refrigerated at-80 ℃.
The culture medium in the S1 is DMEM culture medium.
PEG8000 is selected as PEG in S4.
The concentration of PEG in S4 is 40%.
The operation steps of the inverted absorbent paper method in S4 are as follows:
A. reversely buckling the centrifugal tube which is centrifuged and then supernatant fluid is removed on absorbent paper, and air-drying for 8min at 8 ℃ under the aseptic condition;
B. adding 1.0mL of mixed solution (1:1, V/V) of ethanol and ether into the air-dried bottom precipitate, shaking, centrifuging at 2 deg.C and 1500 Xg for 20min, discarding the supernatant, turning over the centrifuged centrifuge tube on absorbent paper, and air-drying at 8 deg.C under aseptic condition for 10min to obtain bottom precipitate.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (5)
1. A cell supernatant exosome extraction process based on a precipitation reagent method is characterized in that: the method comprises the following steps:
s1, putting cells needing to extract exosomes into a culture medium, and carrying out 3-6% CO treatment at 36-38 DEG C2Culturing the seeds by attaching to the wall for 70-72 d under the aseptic condition;
s2, under the aseptic condition, taking 0.5-1.0 mL of supernatant of the S1 culture medium, placing the supernatant into a centrifuge at the temperature of 2-6 ℃ and 1500-2000 Xg for 10-20 min, discarding the bottom precipitate after centrifugation, and obtaining cell supernatant;
s3, under the aseptic condition, the cell supernatant in the S2 passes through a 0.22 mu m filter to obtain a filtered supernatant;
s4, shaking the filtered supernatant in the S3 and 10-40% of PEG solution uniformly according to the volume ratio of 1: 4-8 under an aseptic condition, standing for 10-12 h at the temperature of 2-6 ℃, centrifuging for 20-30 min at 2500-3000 Xg, discarding the centrifuged supernatant, and obtaining a bottom precipitate by a back-off absorption paper method;
s5, under an aseptic condition, adding 5-7.5 mL of PBS (pH7.2-7.4) into the bottom precipitate in the S3, carrying out heavy suspension, adding 0.5-1.0 mL of 6.0-10.0 mg/mL of streptavidin precipitator, uniformly shaking, and standing for 20-30 min at 2-8 ℃;
s6, centrifuging the liquid obtained after S5 is stood at 2500-3000 Xg for 20-30 min at the temperature of 2-8 ℃ under the aseptic condition, discarding the supernatant, and removing magnetic beads by using a magnetic frame to obtain a precipitate, namely the required exosome;
s7, adding 0.5-1.0 ml PBS solution (pH7.2-7.4) into the exosome in the S6, carrying out heavy suspension, and then refrigerating at-70 to-80 ℃.
2. The process for extracting exosomes from cell supernatant based on the precipitation reagent method according to claim 1, characterized in that: the culture medium in the S1 is one of RPMI1640 culture medium and DMEM culture medium.
3. The process for extracting exosomes from cell supernatant based on the precipitation reagent method according to claim 1, characterized in that: the PEG in the S4 is selected from one of PEG6000 and PEG 8000.
4. The process for extracting exosomes from cell supernatant based on the precipitation reagent method according to claim 1, characterized in that: the concentration of PEG in the S4 is 10-40%.
5. The process for extracting exosomes from cell supernatant based on the precipitation reagent method according to claim 1, characterized in that: the operation steps of the inverted absorbent paper method in S4 are as follows:
A. reversely buckling the centrifugal tube which is centrifuged and then supernatant fluid is removed on absorbent paper, and air-drying for 5-10 min under the aseptic condition of 2-8 ℃;
B. adding 0.5-1.0 mL of ethanol and ether mixed solution (1:1, V/V) into the air-dried bottom sediment, shaking, centrifuging at 2-6 ℃ and 1500-2000 Xg for 10-20 min, discarding the centrifuged supernatant, reversely covering the centrifuged centrifuge tube on absorbent paper, and air-drying at 2-8 ℃ for 5-10 min under an aseptic condition to obtain the bottom sediment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911114046.5A CN110747158A (en) | 2019-11-14 | 2019-11-14 | Cell supernatant exosome extraction process based on precipitation reagent method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911114046.5A CN110747158A (en) | 2019-11-14 | 2019-11-14 | Cell supernatant exosome extraction process based on precipitation reagent method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110747158A true CN110747158A (en) | 2020-02-04 |
Family
ID=69283242
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911114046.5A Pending CN110747158A (en) | 2019-11-14 | 2019-11-14 | Cell supernatant exosome extraction process based on precipitation reagent method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110747158A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112704688A (en) * | 2020-12-02 | 2021-04-27 | 焕生汇生物基因技术(北京)有限公司 | Hair growth injection based on autologous adipose-derived stem cell exosomes and preparation method thereof |
| CN113249302A (en) * | 2021-06-23 | 2021-08-13 | 北京求臻医学检验实验室有限公司 | Efficient exosome separation and purification method |
| CN113502256A (en) * | 2021-06-11 | 2021-10-15 | 广州市第一人民医院（广州消化疾病中心、广州医科大学附属市一人民医院、华南理工大学附属第二医院） | Extraction method of sterile exosomes |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105675774A (en) * | 2016-01-19 | 2016-06-15 | 上海交通大学 | Preparation method of saliva extracellular vesicles and application of saliva extracellular vesicles to molecular diagnosis |
| CN107007541A (en) * | 2017-05-23 | 2017-08-04 | 北京希诺赛尔健康科技推广有限公司 | Application of the excretion body in skin-whitening preparation |
| CN107254430A (en) * | 2017-08-11 | 2017-10-17 | 上海浦美生物医药科技有限公司 | A kind of method based on positive charge adsorbing separation excretion body |
| CN107523536A (en) * | 2017-10-20 | 2017-12-29 | 上海市同济医院 | A kind of extracting method of tissue-derived excretion body and application |
| CN107893051A (en) * | 2017-10-11 | 2018-04-10 | 北京大学 | A kind of method of excretion body in serum using immuno magnetic cell separation |
| CN108148809A (en) * | 2017-09-18 | 2018-06-12 | 兰州大学 | A kind of method that excretion body is detached in the supernatant from tumour cell |
| CN108642004A (en) * | 2018-05-17 | 2018-10-12 | 广东芙金干细胞再生医学有限公司 | A kind of preparation method of excretion body and the stem cell proliferating agent containing excretion body |
| CN108802374A (en) * | 2018-06-25 | 2018-11-13 | 中山大学附属第五医院 | Excretion body nucleic acid detection technique based on magnetic enrichment electrochemical luminescence |
-
2019
- 2019-11-14 CN CN201911114046.5A patent/CN110747158A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105675774A (en) * | 2016-01-19 | 2016-06-15 | 上海交通大学 | Preparation method of saliva extracellular vesicles and application of saliva extracellular vesicles to molecular diagnosis |
| CN107007541A (en) * | 2017-05-23 | 2017-08-04 | 北京希诺赛尔健康科技推广有限公司 | Application of the excretion body in skin-whitening preparation |
| CN107254430A (en) * | 2017-08-11 | 2017-10-17 | 上海浦美生物医药科技有限公司 | A kind of method based on positive charge adsorbing separation excretion body |
| CN108148809A (en) * | 2017-09-18 | 2018-06-12 | 兰州大学 | A kind of method that excretion body is detached in the supernatant from tumour cell |
| CN107893051A (en) * | 2017-10-11 | 2018-04-10 | 北京大学 | A kind of method of excretion body in serum using immuno magnetic cell separation |
| CN107523536A (en) * | 2017-10-20 | 2017-12-29 | 上海市同济医院 | A kind of extracting method of tissue-derived excretion body and application |
| CN108642004A (en) * | 2018-05-17 | 2018-10-12 | 广东芙金干细胞再生医学有限公司 | A kind of preparation method of excretion body and the stem cell proliferating agent containing excretion body |
| CN108802374A (en) * | 2018-06-25 | 2018-11-13 | 中山大学附属第五医院 | Excretion body nucleic acid detection technique based on magnetic enrichment electrochemical luminescence |
Non-Patent Citations (2)
| Title |
|---|
| CHUN-YAN LV等: "A PEG-based method for the isolation of urinary exosomes and its application in renal fibrosis diagnostics using cargo miR-29c and miR-21 analysis", 《INT UROL NEPHROL》 * |
| 贺娇等: "外泌体提取方法及鉴定分析研究进展", 《中华实用诊断与治疗杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112704688A (en) * | 2020-12-02 | 2021-04-27 | 焕生汇生物基因技术(北京)有限公司 | Hair growth injection based on autologous adipose-derived stem cell exosomes and preparation method thereof |
| CN113502256A (en) * | 2021-06-11 | 2021-10-15 | 广州市第一人民医院（广州消化疾病中心、广州医科大学附属市一人民医院、华南理工大学附属第二医院） | Extraction method of sterile exosomes |
| CN113502256B (en) * | 2021-06-11 | 2023-09-19 | 广州市第一人民医院（广州消化疾病中心、广州医科大学附属市一人民医院、华南理工大学附属第二医院） | Extraction method of sterile exosomes |
| CN113249302A (en) * | 2021-06-23 | 2021-08-13 | 北京求臻医学检验实验室有限公司 | Efficient exosome separation and purification method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110747158A (en) | Cell supernatant exosome extraction process based on precipitation reagent method | |
| Taylor et al. | Methods of isolating extracellular vesicles impact down-stream analyses of their cargoes | |
| CN113444682A (en) | Exosome separation and purification method | |
| CN110511902B (en) | Extracellular vesicle separation and enrichment method based on exclusion chromatography and ultrafiltration technology | |
| Chen et al. | Preparation of anionic polyelectrolyte modified magnetic nanoparticles for rapid and efficient separation of lysozyme from egg white | |
| CN106289927A (en) | A kind of from tumor cell supernatant, separate microcapsule bubble and the method for secreting outward body thereof | |
| CN104418928B (en) | A kind of preparation method of 1,4 fourth disulfonic acid ademetionine | |
| CN111426767A (en) | Magnetic nano composite material, preparation thereof and application thereof in food detection | |
| CN109609458A (en) | A kind of method of low speed ultrafiltration centrifugation conjugated polymer sedimentation separation excretion body | |
| CN112108128A (en) | Hydrophilic hyperbranched polyglycidyl ether anion magnetic adsorbent and preparation method and application thereof | |
| CN112048462B (en) | Extracellular vesicle separation and enrichment method based on anionic polymer modified matrix | |
| Ding et al. | Core-shell magnetic zeolite imidazolate framework-8 as adsorbent for magnetic solid phase extraction of brucine and strychnine from human urine | |
| US9119875B2 (en) | Matrix incorporated fluorescent porous and non-porous silica particles for medical imaging | |
| CN111394307A (en) | Method for separating and purifying exosome from plasma and application | |
| CN112877274B (en) | Extracellular vesicle magnetic imprinting material and preparation method and application thereof | |
| CN113388520A (en) | Purification method of extracellular vesicles | |
| CN103333938B (en) | The hepatitis B surface antigen of recombinant Saccharomyces cerevisiae expression and production method, Hepatitis B virus vaccine and production method thereof | |
| WO2023142219A1 (en) | A method for large-scale preparation of high-purity exosomes | |
| CN109136164A (en) | A method of extracellular vesica is separated based on LBL method modification heparin | |
| CN109941976B (en) | Method for preparing nano-selenium by using theabrownin and nano-selenium prepared by method | |
| CN103421764A (en) | Kit for quickly extracting double-stranded RNA of mycovirus and application of kit | |
| CN107304413A (en) | A kind of excretion body quick separating and the kit of purifying | |
| CN115200969A (en) | Method and kit for efficiently separating and purifying plasma exosomes | |
| CN113433196B (en) | Non-denaturing mass spectrometry of membrane protein complexes using nonaethylene glycol monododecyl ether | |
| CN112574950A (en) | Method for extracting cell exosomes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200204 |
|
| WD01 | Invention patent application deemed withdrawn after publication |