CN103900890A - Method for extracting urinary micro vesicle using nanofilm concentration - Google Patents
Method for extracting urinary micro vesicle using nanofilm concentration Download PDFInfo
- Publication number
- CN103900890A CN103900890A CN201410083345.8A CN201410083345A CN103900890A CN 103900890 A CN103900890 A CN 103900890A CN 201410083345 A CN201410083345 A CN 201410083345A CN 103900890 A CN103900890 A CN 103900890A
- Authority
- CN
- China
- Prior art keywords
- urine
- nanometer film
- microcapsule bubble
- rcf
- centrifugal
- 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
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
- External Artificial Organs (AREA)
Abstract
The invention discloses a method for extracting urinary micro vesicle using nanofilm concentration. The method comprises the following steps: firstly, collecting the morning urine of human, performing low speed centrifugation and collecting the upper clear solution; secondly, concentrating the upper clear solution through the nanofilm, dialyzing and filtering to remove moisture and soluble protein and collecting the upper clear solution in the nanofilm; thirdly, performing ultra speed centrifugation, collecting deposition, resuspending with ultra purified water to obtain the urinary micro vesicle. The method for extracting the urinary micro vesicle using nanofilm concentration can rapidly extract urinary micro vesicle in a urine sample, provides accurate information for clinical and basic science researches and brings possibility of realizing rapid and large scale urine treatment.
Description
Technical field
The invention belongs to microcapsule bubble extractive technique field, be specifically related to a kind of method that adopts nanometer film concentration method to extract urine microcapsule bubble.
Background technology
Urine, as a kind of source of undamaged urine protein biology label, more and more causes everybody interest.Urine microcapsule bubble is to be secreted by renal cells, discharges into the vesica of urethra, therefore, likely carries the molecular marked compound of kidney structure and function damage.Protein science method has identified a large amount of high-abundance proteins in urine, and the contained information of urine microcapsule bubble is considered to candidate's biomarker of disease.The exploitation of urine microcapsule bubble protein science contributes to understand the biological function of urinary system urine microcapsule bubble, is more conducive to understand relevant disease pathophysiological mechanism, is also the target spot of finding the urine biomarker in disease incidence mechanism process.
Secreting body (Exosomes) is outward that a class originates from endocytosis system, the microcapsule bubble of the diameter that the mode being merged by plasma membrane by cell multivesicular body is secreted between 40-100nm.Exosomes is found to be in 1981 at first, by Trams etc. in the time that research normal cell and tumour cell come off 5 ' nucleotide excision enzyme active of corpusculum, under Electronic Speculum, find that diameter also has a lot of secondary vesicas in the vesica of 500-1000nm, and the vesica of the about 40nm of diameter.Exosomes is made up of double-layer of lipoid, transmembrane protein and the hydrophilic core, mRNAs and the microRNAs that contain albumen, can serve as intercellular carrier, by cargo carrier function, transmits the information of biological, physiological and pathology.Exosomes participates in intracellular signal transduction.Depend on its origin, exosomes can regulate process of immune regulation, set up tumor escape mechanism and mediation regeneration and degenerative process.Exosomes is the intercellular signal device of molecular complexity, has several functions, is potential clinical diagnosis new tool, potential new disease treatment target spot.
The urine microcapsule bubble extractive technique of report is mainly supercentrifugation at present both at home and abroad.Supercentrifugation is applicable to the processing of urine sample in a small amount, is generally no more than 5mL urine, and the method needs the ultracentrifuge of 200000g, and density gradient centrifugation is expensive, can not disposable processing great amount of samples.
Summary of the invention
The object of the present invention is to provide a kind of method that adopts nanometer film concentration method to extract urine microcapsule bubble, the method can disposable processing great amount of samples, and sample throughput is large, and required instrument relative cost is lower, and treatment scheme is easy.
Above-mentioned purpose of the present invention is achieved through the following technical solutions: a kind of method that adopts nanometer film concentration method to extract urine microcapsule bubble, contains following steps:
(1) low-speed centrifugal, chooses fresh morning specimen, adopt low speed centrifuge, centrifugal after, collect supernatant;
(2) nanometer film is concentrated, the supernatant obtaining in step (1) is poured in nanometer film concentrated, dialysis filtration is removed moisture and soluble protein, in the time that solution filters near nanometer film bottom, add ultrapure water wash-out, further remove the soluble protein in urine, purifying urine microcapsule bubble, collects supernatant;
(3) ultracentrifugation, the supernatant that step (2) is obtained, after adopting ultracentrifuge centrifugal, collects the urinary precipitation of centrifuge tube bottom, obtains urinating microcapsule bubble after resuspended with ultrapure water.
When the middle low-speed centrifugal of step of the present invention (1), the relative centrifugal force(RCF) RCF of hydro-extractor is 2000g, and temperature is room temperature, and centrifugation time is 30min.
First the present invention adopts low-speed centrifugal to remove the foreign material such as cell fragment, bacterium in sample.
The fresh urina sanguinis that the fresh morning specimen adopting in step of the present invention (1) is behaved, does not add antiseptic, and room temperature is placed and is no more than 3 hours, or is stored in 4 ℃ of refrigerators, as needs transportation, need add dry ice.
In step of the present invention (2), in the time that solution preferably filters near 3～5cm place, nanometer film bottom, preferably add 200mL ultrapure water wash-out, further remove the soluble protein in urine, purifying urine microcapsule bubble, preferably collects 5mL supernatant.
Described in step of the present invention (2), the molecular cut off of nanometer film is preferably 1000kDa.
In the time that the molecular cut off of nanometer film adopting is too low, the urine microcapsule bubble purity obtaining is inadequate, contain the soluble protein that molecular weight is larger, the maximum molecular cut off of nanometer film that we take, be chosen as 1000kDa, only have in the time that the molecular cut off of nanometer film is about 1000kDa, in conjunction with low speed centrifuge and ultracentrifuge, can extract purer urine microcapsule bubble.
When ultracentrifuge is centrifugal in step of the present invention (3), the relative centrifugal force(RCF) RCF of hydro-extractor is 40000g at least, and centrifuging temperature is 4 ℃, and centrifugation time is 1～2 hour.
When in step of the present invention (3), ultracentrifuge is centrifugal, the relative centrifugal force(RCF) RCF of hydro-extractor is preferably 40000～100000g, and centrifuging temperature is 4 ℃, and centrifugation time is 1～2 hour.
Compared with prior art, tool of the present invention has the following advantages:
(1) disposal route of the present invention is easy: when supercentrifugation is processed a large amount of urine sample as 50mL, need first by urine vacuum drying to 5mL(be generally original volume 10%), then carry out ultracentrifugation, extract urine microcapsule bubble; When combining nano membrane concentration method utilizes nanometer film dialysis in the present invention, remove a large amount of moisture, convenient and swift;
(2) the inventive method high specificity: inevitably can cause the change of temperature in prior art in supercentrifugation process of vacuum drying, may destroy the film of urine microcapsule bubble, loss part microcapsule bubble;
(3) the inventive method is saved time: each flow process of supercentrifugation institute's spended time that gets off is longer, and processing a sample generally needs the time of 3-5 days, and nanometer film concentration method in the present invention only needs the time of 2 days;
(4) the inventive method economic cost is low: supercentrifugation needs the ultracentrifuge of 200000g, expensive, and nanometer film concentration method in the present invention only needs the hydro-extractor of 40000g, can save economic cost.
Accompanying drawing explanation
Fig. 1 is 40000 times of transmission electron microscope pictures urinating microcapsule bubble in embodiment 1-3;
Fig. 2 is 50000 times of transmission electron microscope pictures urinating microcapsule bubble in embodiment 1-3;
Fig. 3 is the concentration figure of the urine microcapsule bubble that in embodiment 1-3, NanoSight observes, wherein horizontal ordinate representative diameter 0-900nm, and ordinate represents vesica concentration, unit 10
6individual/mL;
Fig. 4 is the video interception of the urine microcapsule bubble that in embodiment 1-3, NanoSight observes, and in figure, white spherical bright spot is vesica;
Fig. 5 is the particle diameter relative intensity figure of the urine microcapsule bubble that in embodiment 1-3, NanoSight observes, and wherein horizontal ordinate is particle diameter 0-900nm, and ordinate is relative intensity;
Fig. 6 is the particle diameter relative intensity three-dimensional plot of the urine microcapsule bubble that in embodiment 1-3, NanoSight observes.
Embodiment
The employing nanometer film concentration method that the present embodiment provides is extracted the method for urine microcapsule bubble, specifically containing following steps:
(1) low-speed centrifugal: get 100mL healthy volunteer's morning specimen, study, low speed centrifuge (Allegra
tMx-22Centrifuge, Beckman Coulter), relative centrifugal force(RCF) (RCF) 2000g, room temperature, 30 minutes, remove the foreign material such as cell fragment, bacterium, collect supernatant;
(2) nanometer film is concentrated: the supernatant that low-speed centrifugal is collected is all poured in nanometer film, and dialysis filtration, in the time that solution filters 3-5cm place, nanometer film bottom, add 200mL ultrapure water wash-out, further remove the soluble protein in urine, purifying urine microcapsule bubble, collects 5mL supernatant;
When wherein nanometer film is dialysed, can adopt easy nanometer film enrichment facility, can use time, oneself is arranged in pairs or groups, and specifically can comprise 1, funnel, nanometer film length 30～35cm, one, 1000mL graduated cylinder, universal film folder (Universal Closures), two of sealed membranes (2cm × 6cm).
Must the Spectrum Labs Inc of pure (Shanghai) trade Co., Ltd purchased from bodyguard about nanometer film, its molecular cut off is 1000kDa.
(3) ultracentrifugation: adopt ultracentrifuge, RCF40000g, 4 ℃, 2 hours, abandon supernatant, collect centrifuge tube bottom urinary precipitation, urinate microcapsule bubble, 1mL ultrapure water is resuspended;
Before ultracentrifugation, preferably utilize analytical balance to carry out trim between two to all samples, guarantee the not axle of breakdown diagnosis balance, (μ g) to require to accomplish to be accurate to after radix point 3.
(4) transmission electron microscope observing: get the urine microcapsule bubble 20 μ L of preparation in step (3), point sample is in copper mesh, after 5 minutes, give 3%(W/V) phosphotungstic acid negative staining 2 minutes, ultrapure water washing twice, dry detection after 5 minutes, regulates transmission electron microscope focal length, observe form and the particle diameter of urine microcapsule bubble, concrete outcome is shown in shown in Fig. 1-2, can find out from Fig. 1-2, and urine microcapsule bubble is evenly distributed, cup-shaped or circular, particle diameter is at 30～100nm.
Before adopting transmission electron microscope observing, require collected urine microcapsule bubble to carry out protein quantification, the urine microcapsule bubble of getting 10 μ g/mL carries out morphological observation.Protein quantification can take Bradford Coomassie brilliant blue method to carry out protein quantification to collected urine microcapsule bubble, gets BSA as standard items, proportional diluted, and drawing standard curve, according to the urine microcapsule bubble protein concentration of typical curve detection testing sample; Measure in triplicate, average.
(5) NanoSight detects: by ultrapure water dilute urine microcapsule bubble, proportional diluted, dilution is 200 times, syringe loading, and each 2～3m, all cleans and detects groove with ultrapure water before and after loading; Adjust NanoSight instrument parameter, to proper focal length, microcapsule bubble is carried out to analytic record.Testing result is shown in shown in Fig. 3-6, from Fig. 3-6, can find out, urinate the particle diameter main peak of microcapsule bubble at 102nm, assorted peak, sample is purer, in Fig. 4, show, in figure, bright spot is allantois bubble, the particle diameter/relative intensity figure of show sample in Fig. 5, and wherein horizontal ordinate is particle diameter 0-900nm, ordinate is relative intensity, and the vesica of visible 100nm particle diameter is in the great majority; Fig. 6 is particle diameter/relative intensity three-dimensional plot of sample, also can find out that the vesica of 100nm particle diameter is in the great majority from 6.
(6) testing result
Under transmission electron microscope, urinate micro-capsule and soak cup-shaped or spherical, be evenly distributed, shown in concrete visible Fig. 1-2.
The particle diameter mean value of urine microcapsule bubble is 102nm, shown in concrete visible Fig. 3.
Urine microcapsule bubble is evenly distributed, and Brownian movement is remarkable.
The quantity of viewed urine microcapsule bubble is 3.8 × 10
8/ mL, due to 200 times of Sample Dilutions, therefore the data of the urine microcapsule bubble of raw sample should be 7.6 × 10
10/ mL.
Embodiment 2
The employing nanometer film concentration method that the present embodiment provides is extracted the method for urine microcapsule bubble, specifically containing following steps:
(1) low-speed centrifugal: get 100mL healthy volunteer's morning specimen, study, low speed centrifuge (Allegra
tMx-22Centrifuge, Beckman Coulter), relative centrifugal force(RCF) (RCF) 2000g, room temperature, 30 minutes, remove the foreign material such as cell fragment, bacterium, collect supernatant;
(2) nanometer film is concentrated: the supernatant that low-speed centrifugal is collected is all poured in nanometer film, and dialysis filtration, in the time that solution filters 3-5cm place, nanometer film bottom, add 200mL ultrapure water wash-out, further remove the soluble protein in urine, purifying urine microcapsule bubble, collects 5mL supernatant;
Must the Spectrum Labs Inc of pure (Shanghai) trade Co., Ltd purchased from bodyguard about nanometer film, its molecular cut off is 1000kDa.
(3) ultracentrifugation: adopt ultracentrifuge, RCF60000g, 4 ℃, 1.5 hours, abandon supernatant, collect centrifuge tube bottom urinary precipitation, urinate microcapsule bubble, 1mL ultrapure water is resuspended;
(4) transmission electron microscope observing: get the urine microcapsule bubble 20 μ L of preparation in step (3), point sample is in copper mesh, after 5 minutes, give 3%(W/V) phosphotungstic acid negative staining 2 minutes, ultrapure water washing twice, dry detection after 5 minutes, regulates transmission electron microscope focal length, observe form and the particle diameter of urine microcapsule bubble, concrete outcome is shown in shown in Fig. 1-2, can find out from Fig. 1-2, and urine microcapsule bubble is evenly distributed, cup-shaped or circular, particle diameter is at 30～100nm.
(5) NanoSight detects: by ultrapure water dilute urine microcapsule bubble, proportional diluted, dilution is 200 times, syringe loading, and each 2～3m, all cleans and detects groove with ultrapure water before and after loading; Adjust NanoSight instrument parameter, to proper focal length, microcapsule bubble is carried out to analytic record.Testing result is shown in shown in Fig. 3-6, from Fig. 3-6, can find out, urinate the particle diameter main peak of microcapsule bubble at 102nm, assorted peak, sample is purer, in Fig. 4, show, in figure, bright spot is allantois bubble, the particle diameter/relative intensity figure of show sample in Fig. 5, and wherein horizontal ordinate is particle diameter 0-900nm, ordinate is relative intensity, and the vesica of visible 100nm particle diameter is in the great majority; Fig. 6 is particle diameter/relative intensity three-dimensional plot of sample, also can find out that the vesica of 100nm particle diameter is in the great majority from 6.
(6) testing result
Under transmission electron microscope, urinate micro-capsule and soak cup-shaped or spherical, be evenly distributed, shown in concrete visible Fig. 1-2.
The particle diameter mean value of urine microcapsule bubble is 102nm, shown in concrete visible Fig. 3.
Urine microcapsule bubble is evenly distributed, and Brownian movement is remarkable.
Embodiment 3
The employing nanometer film concentration method that the present embodiment provides is extracted the method for urine microcapsule bubble, specifically containing following steps:
(1) low-speed centrifugal: get 100mL healthy volunteer's morning specimen, study, low speed centrifuge (Allegra
tMx-22Centrifuge, Beckman Coulter), relative centrifugal force(RCF) (RCF) 2000g, room temperature, 30 minutes, remove the foreign material such as cell fragment, bacterium, collect supernatant;
(2) nanometer film is concentrated: the supernatant that low-speed centrifugal is collected is all poured in nanometer film, and dialysis filtration, in the time that solution filters 3-5cm place, nanometer film bottom, add 200mL ultrapure water wash-out, further remove the soluble protein in urine, purifying urine microcapsule bubble, collects 5mL supernatant;
Nanometer film must the Spectrum Labs Inc of pure (Shanghai) trade Co., Ltd purchased from bodyguard, and its molecular cut off is 1000kDa.
(3) ultracentrifugation: adopt ultracentrifuge, RCF100000g, 4 ℃, 1 hour, abandon supernatant, collect centrifuge tube bottom urinary precipitation, urinate microcapsule bubble, 1mL ultrapure water is resuspended;
(4) transmission electron microscope observing: get the urine microcapsule bubble 20 μ L of preparation in step (3), point sample is in copper mesh, after 5 minutes, give 3%(W/V) phosphotungstic acid negative staining 2 minutes, ultrapure water washing twice, dry detection after 5 minutes, regulates transmission electron microscope focal length, observe form and the particle diameter of urine microcapsule bubble, concrete outcome is shown in shown in Fig. 1-2, can find out from Fig. 1-2, and urine microcapsule bubble is evenly distributed, cup-shaped or circular, particle diameter is at 30～100nm.
(5) NanoSight detects: by ultrapure water dilute urine microcapsule bubble, proportional diluted, dilution is 200 times, syringe loading, and each 2～3m, all cleans and detects groove with ultrapure water before and after loading; Adjust NanoSight instrument parameter, to proper focal length, microcapsule bubble is carried out to analytic record.Testing result is shown in shown in Fig. 3-6, from Fig. 3-6, can find out, urinate the particle diameter main peak of microcapsule bubble at 102nm, assorted peak, sample is purer, in Fig. 4, show, in figure, bright spot is allantois bubble, the particle diameter/relative intensity figure of show sample in Fig. 5, and wherein horizontal ordinate is particle diameter 0-900nm, ordinate is relative intensity, and the vesica of visible 100nm particle diameter is in the great majority; Fig. 6 is particle diameter/relative intensity three-dimensional plot of sample, also can find out that the vesica of 100nm particle diameter is in the great majority from 6.
(6) testing result
Under transmission electron microscope, urinate micro-capsule and soak cup-shaped or spherical, be evenly distributed; Shown in concrete visible Fig. 1-2.
The particle diameter mean value of urine microcapsule bubble is 102nm, shown in concrete visible Fig. 3.
Urine microcapsule bubble is evenly distributed, and Brownian movement is remarkable.
The present invention will be described more than to enumerate specific embodiment.It is pointed out that above embodiment, only for the invention will be further described, does not represent protection scope of the present invention, nonessential modification and adjustment that other people prompting according to the present invention is made, still belong to protection scope of the present invention.
Claims (5)
1. adopt nanometer film concentration method to extract a method for urine microcapsule bubble, it is characterized in that containing following steps:
(1) low-speed centrifugal, chooses fresh morning specimen, adopt low speed centrifuge, centrifugal after, collect supernatant;
(2) nanometer film is concentrated, the supernatant obtaining in step (1) is poured in nanometer film concentrated, dialysis filtration is removed moisture and soluble protein, in the time that solution filters to close nanometer film bottom, add ultrapure water wash-out, further remove the soluble protein in urine, purifying urine microcapsule bubble, collects supernatant;
(3) ultracentrifugation, the supernatant that step (2) is obtained, after adopting ultracentrifuge centrifugal, collects the urinary precipitation of centrifuge tube bottom, obtains urinating microcapsule bubble after resuspended with ultrapure water.
2. employing nanometer film concentration method according to claim 1 is extracted the method for urine microcapsule bubble, it is characterized in that: when the middle low-speed centrifugal of step (1), the relative centrifugal force(RCF) RCF of hydro-extractor is 2000g, and temperature is room temperature, and centrifugation time is 30min.
3. employing nanometer film concentration method according to claim 1 is extracted the method for urine microcapsule bubble, it is characterized in that: the nanometer film that the molecular cut off of the nanometer film adopting in step (2) is 1000kDa.
4. employing nanometer film concentration method according to claim 1 is extracted the method for urine microcapsule bubble, it is characterized in that: when in step (3), ultracentrifuge is centrifugal, the relative centrifugal force(RCF) RCF of hydro-extractor is at least 40000g, and centrifuging temperature is 4 ℃, and centrifugation time is 1～2 hour.
5. employing nanometer film concentration method according to claim 4 is extracted the method for urine microcapsule bubble, it is characterized in that: when in step (3), ultracentrifuge is centrifugal, the relative centrifugal force(RCF) RCF of hydro-extractor is 40000g～100000g, and centrifuging temperature is 4 ℃, and centrifugation time is 1～2 hour.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410083345.8A CN103900890A (en) | 2014-03-07 | 2014-03-07 | Method for extracting urinary micro vesicle using nanofilm concentration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410083345.8A CN103900890A (en) | 2014-03-07 | 2014-03-07 | Method for extracting urinary micro vesicle using nanofilm concentration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103900890A true CN103900890A (en) | 2014-07-02 |
Family
ID=50992353
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410083345.8A Pending CN103900890A (en) | 2014-03-07 | 2014-03-07 | Method for extracting urinary micro vesicle using nanofilm concentration |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103900890A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105716923A (en) * | 2016-04-21 | 2016-06-29 | 华南理工大学 | Preparation method of scanning electron microscope sample for polymer vesicae |
| CN106929467A (en) * | 2017-02-16 | 2017-07-07 | 上海交通大学 | The method and kit of a kind of separating high-purity urine excretion body |
| CN109406252A (en) * | 2018-09-28 | 2019-03-01 | 苏州大学 | A kind of centrifugal ultrafiltration sample processing device and its application method |
| CN109929795A (en) * | 2019-03-22 | 2019-06-25 | 南昌大学第二附属医院 | A kind of improvement extracting method of the outer vesica of urine cellule |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102171574A (en) * | 2008-10-06 | 2011-08-31 | 莫尔豪斯医学院 | Detection of HIV-related proteins in urine |
-
2014
- 2014-03-07 CN CN201410083345.8A patent/CN103900890A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102171574A (en) * | 2008-10-06 | 2011-08-31 | 莫尔豪斯医学院 | Detection of HIV-related proteins in urine |
Non-Patent Citations (2)
| Title |
|---|
| 刑宇洋等: "乳腺癌细胞外泌体的分离与鉴定", 《肿瘤》 * |
| 卢婉: "外泌体的研究进展", 《生命的化学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105716923A (en) * | 2016-04-21 | 2016-06-29 | 华南理工大学 | Preparation method of scanning electron microscope sample for polymer vesicae |
| CN106929467A (en) * | 2017-02-16 | 2017-07-07 | 上海交通大学 | The method and kit of a kind of separating high-purity urine excretion body |
| CN106929467B (en) * | 2017-02-16 | 2021-05-18 | 上海交通大学 | Method and kit for separating high-purity urine exosomes |
| CN109406252A (en) * | 2018-09-28 | 2019-03-01 | 苏州大学 | A kind of centrifugal ultrafiltration sample processing device and its application method |
| CN109929795A (en) * | 2019-03-22 | 2019-06-25 | 南昌大学第二附属医院 | A kind of improvement extracting method of the outer vesica of urine cellule |
| CN109929795B (en) * | 2019-03-22 | 2022-08-19 | 南昌大学第二附属医院 | Improved extraction method of urine small extracellular vesicle |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhao et al. | Microwave Digestion—Vacuum Filtration-Automated Scanning Electron Microscopy as a sensitive method for forensic diatom test | |
| CN109153968A (en) | Extracellular vesica (EV) is separated from biologicfluid sample | |
| CN106399250A (en) | Method and kit for separating exosome | |
| CN114591905B (en) | Method for preparing apoptotic vesicles from human erythrocytes and application of apoptotic vesicles | |
| US11260347B2 (en) | Method and device for separating extracellular vesicles from biological liquids with the aid of cascade ultrafiltration | |
| CN103900890A (en) | Method for extracting urinary micro vesicle using nanofilm concentration | |
| KR20160019475A (en) | Method for separation of sporadic cells from body fluids, and apparatus for carrying out said method | |
| CN107523536A (en) | A kind of extracting method of tissue-derived excretion body and application | |
| CN113388575A (en) | Preparation method of mesenchymal stem cell exosome for skin injury repair | |
| JP6617516B2 (en) | Method for detecting target cells contained in blood sample | |
| CN108801746A (en) | A kind of device of separation and enrichment body fluid components | |
| CN104789525A (en) | Kit for allogeneic peripheral blood mononuclear cell separation in vitro and application method of kit | |
| Welsh et al. | A simple, high-throughput method of protein and label removal from extracellular vesicle samples | |
| JP6617495B2 (en) | Method for detecting tumor cells | |
| CN114540297A (en) | Method for separating mesenchymal stem cell exosomes and analyzing miRNA | |
| US11077147B2 (en) | Acellular biologic composition and method of manufacture | |
| CN111351937A (en) | MMR protein expression deletion detection kit and detection method thereof | |
| CN112094809A (en) | Method for extracting exosome from serum or plasma | |
| CN102965339A (en) | Kit for treating human bone marrow, umbilical cord blood, and peripheral blood cells, and cell treatment method | |
| CN110438061B (en) | Method for separating exosome from fluid shear stress perfusion liquid | |
| CN209485830U (en) | The device of hospital and the home-use extracellular vesica of enrichment urine that can make | |
| CN115651076A (en) | Surface marker of human mesenchymal stem cell-derived apoptotic vesicle and application thereof | |
| CN113234677A (en) | Method for extracting exosome from in-vitro tumor tissue | |
| KR101150840B1 (en) | Composition for aggregating biological sample, method for preparing paraffin block using the same, and method for microscopic observation of specimen sample using the paraffin block | |
| CN111638354A (en) | Immunofluorescence kit for detecting expression of peripheral blood circulating tumor cells E-Cadherin of pancreatic cancer patient |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140702 |