CN106222126A - The extracting method of body is secreted by human body fluid China and foreign countries - Google Patents
The extracting method of body is secreted by human body fluid China and foreign countries Download PDFInfo
- Publication number
- CN106222126A CN106222126A CN201610597323.2A CN201610597323A CN106222126A CN 106222126 A CN106222126 A CN 106222126A CN 201610597323 A CN201610597323 A CN 201610597323A CN 106222126 A CN106222126 A CN 106222126A
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- CN
- China
- Prior art keywords
- body fluid
- human body
- extracting method
- fluid sample
- pbs
- 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.)
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Classifications
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- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
Abstract
The invention discloses a kind of human body fluid China and foreign countries and secrete the extracting method of body, comprise the following steps: prepared by body fluid sample: the most aseptically extract human body fluid, and be diluted with PBS, then tentatively remove the cell component in body fluid and cell debris by centrifugal screening, make body fluid sample standby；Body fluid sample purification: filtered above-mentioned body fluid sample by filter membrane, is removed the cell debris in body fluid and other impurity further, stands 10～15 minutes, leave and take precipitate standby；Secrete outward body to extract: suspended by above-mentioned precipitate PBS, make outer body of secreting be suspended in liquid upper strata, then draw upper strata with aseptic needle tubing and contain the outer liquid secreting body, be placed in 80 DEG C and store for future use.The present invention outer secretes that body extracting method is simple to operate, purification degree is high, it is possible to extracts from human body fluid simply and effectively and outer secretes body, provides the foundation guarantee for follow-up scientific research activity and clinical practice.
Description
Technical field
The present invention relates to biological technical field, secrete the extracting method of body particularly to a kind of human body fluid China and foreign countries.
Background technology
To secrete outward body be the nanoscale lipid encapsulation body structure of a kind of a diameter of 30～100nm, internal package albumen, mRNA
With the material such as microRNA, the almost all kinds of cell including tumor cell, can produce and discharge and outer secrete body.
Studying confirmation recently, the body fluid (blood, saliva, urine, emulsion etc.) of people also contains and secretes outward body, and also foreign study mechanism
Point out, secrete outward body and be probably the seed of cancer Spreading and diffusion.Just it is found the eighties in last century owing to secreting outward body, secretes outward body
Research is scarcely out of swaddling-clothes, and therefore it is known little about it by people, has a lot of characteristic for the people to research and to excavate, and studies
The premise secreting outward body is first it to be extracted from human body fluid.
Summary of the invention
In order to secrete body outside preferably studying, the invention discloses a kind of human body fluid China and foreign countries and secrete the extracting method of body, including
Following steps:
Prepared by a, body fluid sample: the most aseptically extract human body fluid, and be diluted with PBS, so
Tentatively remove the cell component in body fluid and cell debris by centrifugal screening afterwards, make body fluid sample standby；
B, body fluid sample purification: filtered the body fluid sample in step a by filter membrane, removed in body fluid further
Cell debris and other impurity, stand 10～15 minutes, leave and take precipitate standby；
C, secrete body outward and extract: the precipitate PBS of gained in step b is suspended, make outer to secrete body and be suspended in
Liquid upper strata, then draws upper strata with aseptic needle tubing and contains the outer liquid secreting body, be placed in-80 DEG C and store for future use.
As a further improvement on the present invention, described PBS is made up of the material of following parts by weight:
Sodium dihydrogen phosphate 8～12%, disodium hydrogen phosphate 5～10%, potassium chloride 1～3%, pancreatin 0.5～1.5%, surplus
For water.
As a further improvement on the present invention, described filter membrane is the cellulose mixture filter membrane of aperture 0.2～0.4 μm.
The invention has the beneficial effects as follows:
The present invention outer secretes that body extracting method is simple to operate, purification degree is high, it is possible to carry from human body fluid simply and effectively
Take and outer secrete body, provide the foundation guarantee for follow-up scientific research activity and clinical practice.
Accompanying drawing explanation
For the technical scheme being illustrated more clearly that in the embodiment of the present invention, below by use required in embodiment
Accompanying drawing is briefly described, it should be apparent that, the accompanying drawing in describing below is only some embodiments of the present invention, for ability
From the point of view of the those of ordinary skill of territory, on the premise of not paying creative work, it is also possible to obtain other according to these accompanying drawings
Accompanying drawing.
Fig. 1 is that the outer of the present invention secretes body extraction process flow chart.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Describe, it is clear that described embodiment is only a part of embodiment of the present invention rather than whole embodiments wholely.
As it is shown in figure 1, by extract in blood of human body outer secrete body as a example by, the outer extracting method secreting body of the present invention, including
Following steps:
Prepared by a, blood sample: the most aseptically extract blood of human body, and be diluted with PBS, so
Tentatively remove the cell component in blood and cell debris by centrifugal screening afterwards, make blood sample standby；
B, blood sample purification: filtered the blood sample in step a by filter membrane, removed in blood further
Cell debris and other impurity, stand 10～15 minutes, leave and take precipitate standby；
C, secrete body outward and extract: the precipitate PBS of gained in step b is suspended, make outer to secrete body and be suspended in
Liquid upper strata, then draws upper strata with aseptic needle tubing and contains the outer liquid secreting body, be placed in-80 DEG C and store for future use.
Wherein, PBS is made up of the material of following parts by weight:
Sodium dihydrogen phosphate 8～12%, disodium hydrogen phosphate 5～10%, potassium chloride 1～3%, pancreatin 0.5～1.5%, surplus
For water.
Filter membrane is the cellulose mixture filter membrane of aperture 0.2～0.4 μm, to ensure that secreting body passes through filter membrane outward, and
Cell debris and other impurity can be filtered.
The present invention outer secretes that body extracting method is simple to operate, purification degree is high, it is possible to carry from human body fluid simply and effectively
Take and outer secrete body, provide the foundation guarantee for follow-up scientific research activity and clinical practice.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all essences in the present invention
Within god and principle, any modification, equivalent substitution and improvement etc. made, should be included within the scope of the present invention.
Claims (3)
1. the extracting method of body is secreted by human body fluid China and foreign countries, it is characterised in that comprise the following steps:
Prepared by a, body fluid sample: the most aseptically extract human body fluid, and be diluted with PBS, then lead to
Cross centrifugal screening and tentatively remove the cell component in body fluid and cell debris, make body fluid sample standby；
B, body fluid sample purification: filtered the body fluid sample in step a by filter membrane, that removes further in body fluid is thin
Born of the same parents' relic and other impurity, stand 10～15 minutes, leave and take precipitate standby；
C, secrete body outward and extract: the precipitate PBS of gained in step b is suspended, make outer to secrete body and be suspended in liquid
Upper strata, then draws upper strata with aseptic needle tubing and contains the outer liquid secreting body, be placed in-80 DEG C and store for future use.
The extracting method of body is secreted by human body fluid the most according to claim 1 China and foreign countries, it is characterised in that described PBS
It is made up of the material of following parts by weight:
Sodium dihydrogen phosphate 8～12%, disodium hydrogen phosphate 5～10%, potassium chloride 1～3%, pancreatin 0.5～1.5%, surplus is water.
The extracting method of body is secreted by human body fluid the most according to claim 1 China and foreign countries, it is characterised in that described filter membrane is hole
The cellulose mixture filter membrane of footpath 0.2～0.4 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610597323.2A CN106222126A (en) | 2016-07-27 | 2016-07-27 | The extracting method of body is secreted by human body fluid China and foreign countries |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610597323.2A CN106222126A (en) | 2016-07-27 | 2016-07-27 | The extracting method of body is secreted by human body fluid China and foreign countries |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106222126A true CN106222126A (en) | 2016-12-14 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610597323.2A Pending CN106222126A (en) | 2016-07-27 | 2016-07-27 | The extracting method of body is secreted by human body fluid China and foreign countries |
Country Status (1)
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| CN (1) | CN106222126A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108085245A (en) * | 2018-02-11 | 2018-05-29 | 中南大学 | A kind of room temperature isolates and purifies the casing filtering device of 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 |
| CN108251373A (en) * | 2018-02-11 | 2018-07-06 | 徐州医科大学 | The extracting method of mouse brain slices excretion body |
| CN109628277A (en) * | 2019-01-23 | 2019-04-16 | 东南大学 | The separation of excretion in-vivo tumour mark miRNA a kind of and detection system and method |
| CN110736732A (en) * | 2019-12-09 | 2020-01-31 | 南阳理工学院 | Method and device for measuring body fluid drug concentration based on Raman spectrum |
| WO2020147252A1 (en) * | 2019-01-14 | 2020-07-23 | 浙江大学 | APPLICATION OF EXOSOME TβRII PROTEIN AS MARKER IN BREAST CANCER DETECTION KIT PREPARATION |
Citations (3)
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| CN104488850A (en) * | 2014-11-28 | 2015-04-08 | 广州赛莱拉干细胞科技股份有限公司 | Method for preparing exosome freeze-dried powder of human amniotic mesenchymal stem cells |
| CN104894062A (en) * | 2015-05-19 | 2015-09-09 | 暨南大学 | Stem cell exosome patch and preparation method and application thereof |
| CN105505854A (en) * | 2016-01-14 | 2016-04-20 | 上海市第六人民医院 | Acquisition method for exosomes derived from human urinary cells and application |
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2016
- 2016-07-27 CN CN201610597323.2A patent/CN106222126A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104488850A (en) * | 2014-11-28 | 2015-04-08 | 广州赛莱拉干细胞科技股份有限公司 | Method for preparing exosome freeze-dried powder of human amniotic mesenchymal stem cells |
| CN104894062A (en) * | 2015-05-19 | 2015-09-09 | 暨南大学 | Stem cell exosome patch and preparation method and application thereof |
| CN105505854A (en) * | 2016-01-14 | 2016-04-20 | 上海市第六人民医院 | Acquisition method for exosomes derived from human urinary cells and application |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN108085245A (en) * | 2018-02-11 | 2018-05-29 | 中南大学 | A kind of room temperature isolates and purifies the casing filtering device of excretion body |
| CN108251373A (en) * | 2018-02-11 | 2018-07-06 | 徐州医科大学 | The extracting method of mouse brain slices excretion body |
| WO2020147252A1 (en) * | 2019-01-14 | 2020-07-23 | 浙江大学 | APPLICATION OF EXOSOME TβRII PROTEIN AS MARKER IN BREAST CANCER DETECTION KIT PREPARATION |
| CN109628277A (en) * | 2019-01-23 | 2019-04-16 | 东南大学 | The separation of excretion in-vivo tumour mark miRNA a kind of and detection system and method |
| CN109628277B (en) * | 2019-01-23 | 2022-02-01 | 东南大学 | System and method for separating and detecting tumor marker miRNA in exosome |
| CN110736732A (en) * | 2019-12-09 | 2020-01-31 | 南阳理工学院 | Method and device for measuring body fluid drug concentration based on Raman spectrum |
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Application publication date: 20161214 |
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