CN107410287A - Cell-preservation liquid and its application - Google Patents
Cell-preservation liquid and its application Download PDFInfo
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- CN107410287A CN107410287A CN201710706910.5A CN201710706910A CN107410287A CN 107410287 A CN107410287 A CN 107410287A CN 201710706910 A CN201710706910 A CN 201710706910A CN 107410287 A CN107410287 A CN 107410287A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
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Abstract
A kind of cell-preservation liquid of the present invention and its application.The cell-preservation liquid in parts by weight, including following component：60~1000 parts of pH buffer, osmotic pressure maintain 10~150 parts of agent, 4~55 parts of anti-coagulants, 1~30 part of mucolytic agent, 800~5000 parts of fixative, component A1~150 part, component B10~30 part, 10000~20000 parts of water；Wherein, the component A is that weight ratio is 1~10：2~110 Sodium azide and lauryl sodium sulfate；The component B is carbon disulfide.The cell-preservation liquid of the present invention has preferably impurity treatment effect, reduces the disturbing factor in cell sample, while target cell therein can keep the feature of its structural form in the long period, be easy to the medical science of target cell to test and analyze, and cost is low.
Description
Technical field
The present invention relates to medical science, more particularly to cell-preservation liquid and its application.
Background technology
As the screening method of cervical lesionses, the materials that are related to during traditional Pap smear, tablet quality, cell are applied
The factors such as separation all can seriously reduce the positive rate of detection.Cytology expert is proposed a kind of sectioning cells diagnosis side
Method --- ThinPrep liquid-based cytology test technology (Thinprep Cytology Test；Abbreviation TCT), it mainly passes through technical office
Reason loses non-diagnostic impurity, makes and is easy to observation clearly thin-layer cell piece, makes diagosis person be more easy to observe, diagnostic accuracy is obvious
Improve.
TCT technologies can preserve significant cell to greatest extent, eliminate artificial loss and destruction, sample cell is consolidated
It is fixed to terminate any ongoing biochemical reaction in the solution, increase the mechanical strength and stability of sample, as far as possible to connect
The native state of nearly biomaterial preserves sample, and picture is apparent, diagnosis is more accurate, eliminates blood, mucus and artificial
Influence of the film-making process to cell.
The cell-preservation liquid of liquid-based is the main consumptive material of ThinPrep liquid-based cytology test technology.Cell in cell-preservation liquid
Thin smear is made in programmed processing, grab sample, and in thin smear, abnormal cell is easily observed and moist
Nuclear structures are apparent, are easy to differentiate, can detect more rudimentary lesion and some more serious lesions, are current states
Popular new technology on border.
If human body living cells departs from original living environment and placed, the physiological function of cell, biological activity and intrinsic
Cellular morphology and structure will change.Existing cell-preservation liquid is often with 40~50% alcohols, physiological saline or to thin
Born of the same parents' sample carries out freezen protective, and the cell holding time of these three methods is very short and has the harm of damage to cell, can cause
Cellular prion protein changes, so as to influence follow-up work.
Different cytology is needed to preserve liquid in addition, different clinical cytology samples makes, to uterine neck brushing piece, oesophagus brush
The clinical cytology sample such as piece, sputum, urine, Pleural effusions, cell-preservation liquid have different requirements.For being mixed with more thin of blood
Born of the same parents imitate this, either conventional or liquid based cytology film-making, and visible more red blood cell under mirror, background dyes red deeply, no
Searched beneficial to cancer cell, directly affect diagnosis.And for the more clinical cytology sample of mucus, particularly cervical smear, oesophagus
Scraping blade is, it is necessary to which specific aim removes the interference of mucus.
The defects of current cell-preservation liquid generally existing is expensive, the cell holding time is short, particularly for blood
During the processing of liquid or the more clinical cytology sample of mucus, sample impurity treatment is undesirable, and the interference such as haemocyte, mucus is thin
The processing of born of the same parents' film-making factor also complies with one's wishes not to the utmost.
The content of the invention
Based on this, it is necessary to provide one kind while effectively protection target cell form, impurity treatment effect is good, interference
Factor is few, and the cell-preservation liquid that the holding time is long, cost is low.
A kind of cell-preservation liquid, in parts by weight, including following component：
Wherein, the component A is that weight ratio is 1~10：2~110 Sodium azide and lauryl sodium sulfate；The component
B is carbon disulfide.
In one of the embodiments, the component A is that weight ratio is 1~10：2~15 Sodium azide and dodecyl sulphur
Sour sodium.
In one of the embodiments, the mucolytic agent be methyl cysteine, ACETYLCYSTEINE,
One or more in dithiothreitol (DTT) (DTT, a kind of protein reducing agent), dihydroxy dithiothreitol (DTT).Preferably two sulphur are revived
Sugar alcohol.
In one of the embodiments, the fixative is in methanol, ethanol, ethylene glycol, isopropanol, formaldehyde, glacial acetic acid
One or more.
In one of the embodiments, the fixative is that weight ratio is 2~25：750~5000：10~155 methanol,
The mixed liquor of isopropanol and 35~40% (volumetric concentration) formalins.
In one of the embodiments, described pH buffer is Tris-HCl buffer solutions, PIPES, phosphate buffer
At least one of (PBS), it is 6.5~7.5, preferably 7.0~7.4 to maintain the pH value of cell-preservation liquid.Phosphate delays
Fliud flushing can be at least one of sodium phosphate and potassium phosphate, preferably kaliumphosphate buffer.
In one of the embodiments, it is in sodium chloride, potassium chloride, calcium chloride, sodium acid carbonate that the osmotic pressure, which maintains agent,
One or more.For maintaining the ionic strength in cell-preservation liquid, the stability of target cell is kept.Preferably chlorination
Sodium.
In one of the embodiments, described anti-coagulants is at least one of EDTA, heparin.Preferably EDTA.
In one of the embodiments, described cell-preservation liquid, in parts by weight, including following component：
Cell-preservation liquid answering in cervical smear, oesophagus scraping blade or endometrium scraping blade described in the offer of the invention
With.
Compared with prior art, the invention has the advantages that：
Cell-preservation liquid of the present invention：First, using specified weight than Sodium azide and lauryl sodium sulfate as
Component A, wherein, the cell for the cytode (such as red blood cell) that lauryl sodium sulfate can be destroyed easily in cell sample
Film, make wherein hemoglobin dissolution, while mucus can also be decomposed, can be to having further combined with the use of Sodium azide
Nucleus forms protection, prevents microorganism in remaining component or environment etc. from producing destruction to target cell；Secondly, with curing
Carbon can decompose to the hemoglobin of dissolution as component B, reduce it and target cell is disturbed, while carbon disulfide be made
For specific enzyme inhibitor, the metabolism of target cell can be reduced, is advantageous to protect the morphosis of target cell.
On this basis, distributing rationally with reference to each component parts by weight, cell-preservation liquid of the invention have preferably miscellaneous
Matter treatment effect, reduce the disturbing factor in cell sample, while target cell therein can be kept in the long period
The feature of its structural form, it is easy to the medical science of target cell to test and analyze, and cost is low.
Particular kind of mucolytic agent is further used, the reduction decomposition of mucus can be promoted, interference is reduced, with reference to institute
Fixative is stated, the cell membrane of target cell more can be quickly passed through, biomolecule therein is fixed, is advantageous to mesh
Mark the holding of cellular prion protein.
Brief description of the drawings
Fig. 1 is liquid A1 mounting observation result figure to be preserved in embodiment (left side is to observe for 0 month, and right side is sees after placing 2 months
Examine)；
Fig. 2 is liquid A2 mounting observation result figure to be preserved in embodiment (left side is to observe for 0 month, and right side is sees after placing 2 months
Examine)；
Fig. 3 is liquid A3 mounting observation result figure to be preserved in embodiment (left side is to observe for 0 month, and right side is sees after placing 2 months
Examine)；
Fig. 4 is liquid B1 mounting observation result figure to be preserved in embodiment (left side is to observe for 0 month, and right side is sees after placing 2 months
Examine)；
Fig. 5 is liquid B2 mounting observation result figure to be preserved in embodiment (left side is to observe for 0 month, and right side is sees after placing 2 months
Examine)；
Fig. 6 be in embodiment contrast preserve liquid 1 mounting observation result figure (left side be 0 month observe, right side for place 2 months
Observe afterwards)；
(left side is to be observed after placing 2 months with 10x to the mounting observation result figure that Fig. 7 is preservation liquid C1 in embodiment, and right side is
Observed after placing 2 months with 40x)；
(left side is to be observed after placing 2 months with 10x to the mounting observation result figure that Fig. 8 is preservation liquid C2 in embodiment, and right side is
Observed after placing 2 months with 40x)；
(left side is to be observed after placing 2 months with 10x to the mounting observation result figure that Fig. 9 is preservation liquid C3 in embodiment, and right side is
Observed after placing 2 months with 40x)；
Figure 10 be in embodiment contrast preserve liquid 2 mounting observation result figure (left side be place 2 months after observed with 10x, the right side
Side is to be observed after placing 2 months with 40x).
Embodiment
The cell-preservation liquid of the present invention and its application are described in further detail below in conjunction with specific embodiment.
Embodiment 1
Three kinds of cell-preservation liquids of the present embodiment, including following component (table 1)：
Table 1
| Preserve liquid A1 | Preserve liquid A2 | Preserve liquid A3 | |
| Disodium hydrogen phosphate | 580.2g | 580.2g | 580.2g |
| Sodium dihydrogen phosphate | 59.28g | 59.28g | 59.28g |
| Sodium chloride | 100g | 10 | 150 |
| EDTA | 40g | 4 | 55 |
| Sodium azide | 4g | 4g | 4g |
| Dithiothreitol (DTT) DTT | 15g | 30 | 1 |
| Methanol | 20mL(15.8g) | 2.5mL(2.0g) | 30mL(23.7g) |
| Isopropanol | 4000mL(3160g) | 1000mL(790g) | 6000mL(4740g) |
| 40% formaldehyde | 100mL(108.3g) | 12.5mL(13.5g) | 140mL(151.6g) |
| Lauryl sodium sulfate | 12.6g | 12.6g | 12.6g |
| Carbon disulfide | 20g | 10g | 30g |
| Purified water | Add water to 20L | Add water to 20L | Add water to 20L |
Each component is mixed according to the dosage of table 1, produces the cell-preservation liquid.
Embodiment 2
Three kinds of cell-preservation liquids of the present embodiment, including following component (table 2)：
Table 2
Each component is mixed according to the dosage of table 2, produces the cell-preservation liquid.
Embodiment 3
Four kinds of cell-preservation liquids of the present embodiment, including following component (table 3)：
Table 3
Each component is mixed according to the dosage of table 3, produces the cell-preservation liquid.
Using above-mentioned each cell-preservation liquid processing with leukorrhea/cervical exfoliated cell of batch, the automatic film-makings of HQ TCT are used
Mechanism piece, dyed using HQ pap stainings liquid, mounting observation, as a result as shown in table 4.
Table 4
The most long holding time：Added after referring to sampling, in cell sample and preserve liquid, detected again after can placing 2 months.
General existing cell-preservation liquid, can only be placed 3 weeks, once more than 1 month, sample can not just reuse.
Each technical characteristic of embodiment described above can be combined arbitrarily, to make description succinct, not to above-mentioned reality
Apply all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, the scope that this specification is recorded all is considered to be.
Embodiment described above only expresses the several embodiments of the present invention, and its description is more specific and detailed, but simultaneously
Can not therefore it be construed as limiting the scope of the patent.It should be pointed out that come for one of ordinary skill in the art
Say, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
- A kind of 1. cell-preservation liquid, it is characterised in that in parts by weight, including following component：Wherein, the component A is that weight ratio is 1~10：2~110 Sodium azide and lauryl sodium sulfate；The component B is Carbon disulfide.
- 2. cell-preservation liquid according to claim 1, it is characterised in that the component A is that weight ratio is 1~10：2~15 Sodium azide and lauryl sodium sulfate.
- 3. cell-preservation liquid according to claim 1, it is characterised in that the mucolytic agent be methyl cysteine, One or more in ACETYLCYSTEINE, dithiothreitol (DTT), dihydroxy dithiothreitol (DTT).
- 4. cell-preservation liquid according to claim 1, it is characterised in that the fixative be methanol, ethanol, ethylene glycol, One or more in isopropanol, formaldehyde, glacial acetic acid.
- 5. cell-preservation liquid according to claim 4, it is characterised in that the fixative is that weight ratio is 2~25：750 ~5000：The mixed liquor of 10~155 methanol, isopropanol and 35~40% formalins.
- 6. cell-preservation liquid according to claim 1, it is characterised in that described pH buffer is Tris-HCl bufferings At least one of liquid, PIPES buffers, phosphate buffer, to maintain the pH value of cell-preservation liquid as 6.5~7.5.
- 7. cell-preservation liquid according to claim 1, it is characterised in that it is sodium chloride, chlorination that the osmotic pressure, which maintains agent, One or more in potassium, calcium chloride, sodium acid carbonate.
- 8. cell-preservation liquid according to claim 1, it is characterised in that described anti-coagulants is EDTA, in heparin extremely Few one kind.
- 9. according to the cell-preservation liquid described in claim any one of 1-8, it is characterised in that in parts by weight, including such as the following group Point：
- 10. the cell-preservation liquid described in claim any one of 1-9 is in cervical smear, oesophagus scraping blade or endometrium scraping blade Using.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710706910.5A CN107410287A (en) | 2017-08-17 | 2017-08-17 | Cell-preservation liquid and its application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710706910.5A CN107410287A (en) | 2017-08-17 | 2017-08-17 | Cell-preservation liquid and its application |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108094407A (en) * | 2017-12-29 | 2018-06-01 | 浙江今复康生物科技有限公司 | A kind of preparation method of cell-preservation liquid and Phlegm Cells reference material |
| CN109042630A (en) * | 2018-09-27 | 2018-12-21 | 苏州市奥健医卫用品有限公司 | A kind of cell-preservation liquid and preparation method thereof |
| CN109090104A (en) * | 2018-09-27 | 2018-12-28 | 广州新诚生物科技有限公司 | Save liquid and its preparation method and application |
| CN109682971A (en) * | 2018-12-18 | 2019-04-26 | 杭州海世嘉生物科技有限公司 | A kind of preservation liquid and its preparation process, application method |
| CN110463686A (en) * | 2019-07-08 | 2019-11-19 | 深圳市华晨阳科技有限公司 | A kind of cell-preservation liquid that can effectively save cell for a long time |
| CN110742059A (en) * | 2019-11-27 | 2020-02-04 | 武汉医尔特科技有限公司 | Liquid-based cell preservation solution and preparation method thereof |
| CN111238893A (en) * | 2020-01-19 | 2020-06-05 | 湖北泰康医疗设备有限公司 | Method for extracting humoral cells for detecting lung cancer |
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
| CN111449056A (en) * | 2020-05-25 | 2020-07-28 | 山西汉济堂医疗科技有限公司 | Novel cell preservation solution and preparation method thereof |
| CN112544609A (en) * | 2020-11-10 | 2021-03-26 | 广州市金圻睿生物科技有限责任公司 | Cervical cell sample preservation solution and application thereof |
-
2017
- 2017-08-17 CN CN201710706910.5A patent/CN107410287A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108094407A (en) * | 2017-12-29 | 2018-06-01 | 浙江今复康生物科技有限公司 | A kind of preparation method of cell-preservation liquid and Phlegm Cells reference material |
| CN109042630A (en) * | 2018-09-27 | 2018-12-21 | 苏州市奥健医卫用品有限公司 | A kind of cell-preservation liquid and preparation method thereof |
| CN109090104A (en) * | 2018-09-27 | 2018-12-28 | 广州新诚生物科技有限公司 | Save liquid and its preparation method and application |
| CN109682971A (en) * | 2018-12-18 | 2019-04-26 | 杭州海世嘉生物科技有限公司 | A kind of preservation liquid and its preparation process, application method |
| CN110463686A (en) * | 2019-07-08 | 2019-11-19 | 深圳市华晨阳科技有限公司 | A kind of cell-preservation liquid that can effectively save cell for a long time |
| CN110742059A (en) * | 2019-11-27 | 2020-02-04 | 武汉医尔特科技有限公司 | Liquid-based cell preservation solution and preparation method thereof |
| CN111238893A (en) * | 2020-01-19 | 2020-06-05 | 湖北泰康医疗设备有限公司 | Method for extracting humoral cells for detecting lung cancer |
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
| CN111449056A (en) * | 2020-05-25 | 2020-07-28 | 山西汉济堂医疗科技有限公司 | Novel cell preservation solution and preparation method thereof |
| CN112544609A (en) * | 2020-11-10 | 2021-03-26 | 广州市金圻睿生物科技有限责任公司 | Cervical cell sample preservation solution and application thereof |
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