CN101999344A - In-vitro tissue preserving fluid and preparation method thereof - Google Patents
In-vitro tissue preserving fluid and preparation method thereof Download PDFInfo
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- CN101999344A CN101999344A CN201010601545XA CN201010601545A CN101999344A CN 101999344 A CN101999344 A CN 101999344A CN 201010601545X A CN201010601545X A CN 201010601545XA CN 201010601545 A CN201010601545 A CN 201010601545A CN 101999344 A CN101999344 A CN 101999344A
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Abstract
The invention discloses in-vitro tissue preserving fluid and a preparation method thereof. Each 1,000 milliliters of preserving fluid comprises the following components by weight: 5 to 200 grams of osmotic pressure regulator, 0.2 to 40 grams of a pH value buffer pair, 3 to 15 grams of potassium chloride, 1 to 5 grams of sodium chloride, 10 to 300 milligrams of antibiotic, 5 to 100 grams of glucose and the balance of deionized water. In the invention, a non-CO2 dependent buffer pair is used to reduce the dependence on CO2 in transport and an air exposure process and reduce the fluctuations of the pH value; a hypertonic and high-potassium environment is used to reduce the swelling of cells and tissues and improve the activities of the cells and tissues; and the antibiotic used is free from causing hypersensitivity reaction. The preparation method is simple and in-vitro tissue preserving fluid has a remarkable in-vitro tissue preserving effect. The in-vitro tissue preserving fluid is low in cost, has low transport requirements, and can be stored at room temperature and conveniently.
Description
Technical field
The present invention relates to preservation liquid medically and preparation method thereof, especially a kind of preservation liquid and compound method thereof of the human or animal tissues that is used to exsomatize.
Background technology
Organ transplant is the major progress of 20th century physianthropy science, has begun since clinical practice from the sixties in 20th century, and transplantation medicine has obtained the development of advancing by leaps and bounds.Yet any clinical organ transplant at first will have high-quality donor, and this is the prerequisite and the basic guarantee of organ transplant success, therefore, being kept at this significance arranged of organ, it is the foundation stone of organ transplant.Closely during the last ten years, Chinese organ transplant quantity has leapt to the first place, Asia, but clinically except kidney is preserved, liquid is preserved in the preservation of organs such as liver, the heart and pancreas still dependence on import.The import organ preservative fluid costs an arm and a leg, for the patient who carries out organ transplant has increased heavy financial burden.
Biological therapy and stem-cell therapy also are the great focuses of current medical domain.Along with the development of biological therapy and stem-cell research, people need to preserve, transport the portion of tissue of donor, and arrival is used for scientific research and treatment behind the destination, owing in the transportation a lot of uncontrollable conditions are arranged, causes tissue and cell in transportation mesometamorphism, death.Import organ preservative fluid preservation effect is better relatively, but also exists transportation environment is had relatively high expectations, and shortcoming such as cost an arm and a leg.Therefore, the urgent need development is a kind of can be used for multiple tissue preservation, and the low cheapness of environmental requirement is preserved liquid to external world.In addition, because penicillin causes allergy easily, and streptomycin has genotoxic potential to human body, and the treatment that the tissue of preserving is applied to human diseases later on is potentially dangerous, and therefore preserves to add antibiotic in the liquid and should avoid adding this two classes antibiotic as far as possible.
Summary of the invention
Technical problem to be solved by this invention is, preservation liquid of a kind of cheap, relative better preserve in vitro tissue of preservation effect and preparation method thereof is provided.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is: a kind of preservation liquid of preserving in vitro tissue, every 1000ml are preserved liquid and are made up of following components by weight ratio:
Osmotic pressure regulator 5～200g
PH value buffering is to 0.2～40g
Potassium chloride 3～15g
Sodium chloride 1～5g
Antibiotic 10～300mg
Glucose 5～100g
The deionized water surplus.
Described osmotic pressure regulator is selected HES or D-40 for use.
The preferred D-40 of described osmotic pressure regulator.
Described pH value buffering is to being selected from a pair of in sodium dihydrogen phosphate/sodium hydrogen phosphate, potassium dihydrogen phosphate/dipotassium hydrogen phosphate, potassium dihydrogen phosphate/sodium hydroxide, sodium dihydrogen phosphate/sodium hydroxide or the sodium hydrogen phosphate/potassium dihydrogen phosphate.
Described pH value buffering is to preferably phosphoric acid potassium dihydrogen/dipotassium hydrogen phosphate, and the amount that wherein every 1000ml preserves liquid phosphoric acid potassium dihydrogen is 0.1～20g, and the amount of dipotassium hydrogen phosphate is 0.1～20g.
Described antibiotic is not for there being the antibiotic that causes hypersensitivity.
Described antibiotic preferably sulfuric acid gentamicin.
The preparation method of the preservation liquid of aforesaid preservation in vitro tissue may further comprise the steps:
(1) the preparation osmotic pressure regulator deionized water aqueous solution stirs and spends the night to dissolving fully, the removal of impurity of 50um membrane filtration;
(2) add the residue solute in the above-mentioned aqueous solution, be stirred to dissolving fully;
(3) pass through 50um successively, 0.4nm, 0.2um, the 0.1um membrane filtration, the filtrate room temperature preservation is standby.
Beneficial effect of the present invention: use the in vitro tissue of this method preparation to preserve liquid, can obtain and advance
The preservation effect that the UW organ preservative fluid is close.There is not obvious cytotoxicity, at room temperature stable in properties.It is little to the potential danger of clinical practice in future not have the antibiotic that causes hypersensitivity.Preparation method of the present invention is simple, and is good for the in vitro tissue preservation effect.Cost is low, and is low for movement requirement, but room temperature preservation, and it is convenient to store.
Embodiment
The preservation liquid of preservation in vitro tissue of the present invention, every 1000ml are preserved liquid and are made up of following components by weight ratio:
Osmotic pressure regulator 5～200g
PH value buffering is to 0.2～40g
Potassium chloride 3～15g
Sodium chloride 1～5g
Antibiotic 10～300mg
Glucose 5～100g
The deionized water surplus.
Described osmotic pressure regulator is selected HES or D-40 for use.
The preferred D-40 of described osmotic pressure regulator.
Described pH value buffering is to being selected from a pair of in sodium dihydrogen phosphate/sodium hydrogen phosphate, potassium dihydrogen phosphate/dipotassium hydrogen phosphate, potassium dihydrogen phosphate/sodium hydroxide, sodium dihydrogen phosphate/sodium hydroxide or the sodium hydrogen phosphate/potassium dihydrogen phosphate.
Described pH value buffering is to preferably phosphoric acid potassium dihydrogen/dipotassium hydrogen phosphate, and the amount that wherein every 1000ml preserves liquid phosphoric acid potassium dihydrogen is 0.1～20g, and the amount of dipotassium hydrogen phosphate is 0.1～20g.
Described antibiotic is not for there being the antibiotic that causes hypersensitivity.
Described antibiotic preferably sulfuric acid gentamicin.
The preparation method of the preservation liquid of aforesaid preservation in vitro tissue may further comprise the steps:
(1) the preparation osmotic pressure regulator deionized water aqueous solution stirs and spends the night to dissolving fully, the removal of impurity of 50um membrane filtration;
(2) add the residue solute in the above-mentioned aqueous solution, be stirred to dissolving fully;
(3) pass through 50um successively, 0.4nm, 0.2um, the 0.1um membrane filtration, the filtrate room temperature preservation is standby.
Below, the invention will be further described in conjunction with specific embodiments, but the present invention is not limited to following embodiment.
Prescription: 1000ml preserves liquid and contains
D-40-40 100g
Potassium dihydrogen phosphate 0.2g
Dipotassium hydrogen phosphate 1.4g
Potassium chloride 7.6g
Sodium chloride 2.2g
Glucose 50g
Gentamicin sulphate 50mg
Compound method:
1, preparation D-40-40 aqueous solution: take by weighing D-40-40100g, add the 800ml deionized water, continuous stirring is spent the night to dissolving fully.Be settled to 1000ml.The removal of impurity of 50um membrane filtration.
2, will add potassium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium chloride, sodium chloride, glucose, gentamicin sulphate according to the above ratio in D-40-40 aqueous solution that prepare, be stirred to dissolving fully.
3, pass through 50um successively, 0.4nm, 0.2um, packing behind the 0.1um membrane filtration.0.2um 0.1um should operate in hundred grades of environment.Room temperature preservation is standby.
The present invention uses non-CO2 dependence buffering right, to reduce in the process that reaches in transit with air exposure for the dependence of CO2, reduces the fluctuation of pH value; Use height to ooze and high potassium environment, reduce the swelling of cell and tissue, strengthen its vigor; Use and do not have the potential hazard of antibiotic minimizing that causes hypersensitivity using in the future.
In sum, among the embodiment that content of the present invention is not confined to, those skilled in the art can propose other embodiment within technological guidance's thought of the present invention, but this embodiment comprises within the scope of the present invention.
Claims (8)
1. preservation liquid of preserving in vitro tissue is characterized in that: every 1000ml preserves liquid and is made up of following components by weight ratio:
Osmotic pressure regulator 5～200g
PH value buffering is to 0.2～40g
Potassium chloride 3～15g
Sodium chloride 1～5g
Antibiotic 10～300mg
Glucose 5～100g
The deionized water surplus.
2. the preservation liquid of preservation in vitro tissue according to claim 1 is characterized in that: described osmotic pressure regulator is selected HES or D-40 for use.
3. the preservation liquid of preservation in vitro tissue according to claim 2 is characterized in that: described osmotic pressure regulator is a D-40.
4. the preservation liquid of preservation in vitro tissue according to claim 1 is characterized in that: described pH value buffering is to being selected from a pair of in sodium dihydrogen phosphate/sodium hydrogen phosphate, potassium dihydrogen phosphate/dipotassium hydrogen phosphate, potassium dihydrogen phosphate/sodium hydroxide, sodium dihydrogen phosphate/sodium hydroxide or the sodium hydrogen phosphate/potassium dihydrogen phosphate.
5. the preservation liquid of preservation in vitro tissue according to claim 4, it is characterized in that: described pH value buffering is to being potassium dihydrogen phosphate/dipotassium hydrogen phosphate, the amount that wherein every 1000ml preserves liquid phosphoric acid potassium dihydrogen is 0.1～20g, and the amount of dipotassium hydrogen phosphate is 0.1～20g.
6. the preservation liquid of preservation in vitro tissue according to claim 1 is characterized in that: described antibiotic is not for there being the antibiotic that causes hypersensitivity.
7. the preservation liquid of preservation in vitro tissue according to claim 6 is characterized in that: described antibiotic is a gentamicin sulphate.
8. as the preparation method of the preservation liquid of the described preservation in vitro tissue of claim 1～7, it is characterized in that, may further comprise the steps:
(1) the preparation osmotic pressure regulator deionized water aqueous solution stirs and spends the night to dissolving fully, the removal of impurity of 50um membrane filtration;
(2) add the residue solute in the above-mentioned aqueous solution, be stirred to dissolving fully;
(3) pass through 50um successively, 0.4nm, 0.2um, the 0.1um membrane filtration, the filtrate room temperature preservation is standby.
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| CN 201010601545 CN101999344B (en) | 2010-12-27 | 2010-12-27 | In-vitro tissue preserving fluid and preparation method thereof |
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| CN 201010601545 CN101999344B (en) | 2010-12-27 | 2010-12-27 | In-vitro tissue preserving fluid and preparation method thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102334472A (en) * | 2011-08-02 | 2012-02-01 | 江苏省北科生物科技有限公司 | Umbilical cord preserving fluid and preparation method thereof |
| CN104673717A (en) * | 2015-02-12 | 2015-06-03 | 中国医科大学 | Biological tissue RNA (ribonucleic acid) protection reagent as well as preparation method and application thereof |
| CN105028388A (en) * | 2015-07-08 | 2015-11-11 | 深圳爱生再生医学科技有限公司 | Preserving fluid and preparation method therefor |
| CN106417257A (en) * | 2016-10-13 | 2017-02-22 | 北京焕生汇生物技术研究院 | Novel in-vitro tissue preserving fluid |
| CN106417254A (en) * | 2016-09-30 | 2017-02-22 | 广州赛莱拉干细胞科技股份有限公司 | Tooth specimen preservation liquid and application thereof and tooth specimen preservation method |
| CN106620763A (en) * | 2016-09-30 | 2017-05-10 | 广州赛莱拉干细胞科技股份有限公司 | Method for sterilizing tooth sample |
| CN107183010A (en) * | 2017-06-26 | 2017-09-22 | 苏州卡睿知光电科技有限公司 | A kind of in vitro tissue preserves liquid |
| CN107318826A (en) * | 2017-06-26 | 2017-11-07 | 苏州卡睿知光电科技有限公司 | A kind of in vitro tissue preserves liquid |
| CN108338160A (en) * | 2018-04-27 | 2018-07-31 | 张长水 | A kind of hair follicle preserves liquid and preparation method thereof |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102334472A (en) * | 2011-08-02 | 2012-02-01 | 江苏省北科生物科技有限公司 | Umbilical cord preserving fluid and preparation method thereof |
| CN102334472B (en) * | 2011-08-02 | 2013-04-10 | 江苏省北科生物科技有限公司 | Umbilical cord preserving fluid and preparation method thereof |
| CN104673717A (en) * | 2015-02-12 | 2015-06-03 | 中国医科大学 | Biological tissue RNA (ribonucleic acid) protection reagent as well as preparation method and application thereof |
| CN104673717B (en) * | 2015-02-12 | 2017-11-28 | 中国医科大学 | A kind of biological tissue RNA protections reagent and its preparation method and application |
| CN105028388A (en) * | 2015-07-08 | 2015-11-11 | 深圳爱生再生医学科技有限公司 | Preserving fluid and preparation method therefor |
| CN106417254A (en) * | 2016-09-30 | 2017-02-22 | 广州赛莱拉干细胞科技股份有限公司 | Tooth specimen preservation liquid and application thereof and tooth specimen preservation method |
| CN106620763A (en) * | 2016-09-30 | 2017-05-10 | 广州赛莱拉干细胞科技股份有限公司 | Method for sterilizing tooth sample |
| CN106417257A (en) * | 2016-10-13 | 2017-02-22 | 北京焕生汇生物技术研究院 | Novel in-vitro tissue preserving fluid |
| CN107183010A (en) * | 2017-06-26 | 2017-09-22 | 苏州卡睿知光电科技有限公司 | A kind of in vitro tissue preserves liquid |
| CN107318826A (en) * | 2017-06-26 | 2017-11-07 | 苏州卡睿知光电科技有限公司 | A kind of in vitro tissue preserves liquid |
| CN108338160A (en) * | 2018-04-27 | 2018-07-31 | 张长水 | A kind of hair follicle preserves liquid and preparation method thereof |
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