CN105918308A - Lung perfusion fluid for perfusing transplant donor lung and preparation method for lung perfusion fluid - Google Patents
Lung perfusion fluid for perfusing transplant donor lung and preparation method for lung perfusion fluid Download PDFInfo
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- CN105918308A CN105918308A CN201610277886.3A CN201610277886A CN105918308A CN 105918308 A CN105918308 A CN 105918308A CN 201610277886 A CN201610277886 A CN 201610277886A CN 105918308 A CN105918308 A CN 105918308A
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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
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
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Abstract
The invention provides lung perfusion fluid for perfusing a transplant donor lung and a preparation method for the lung perfusion fluid. The lung perfusion fluid can be used for providing good lung protecting and resuscitating actions and can meet the requirements of the transplant donor lung on perfusion and preservation; meanwhile, the time of perfusion is relatively long, and the donor lung still can retain good lung compliance and oxygenation capability 6 hours after the donor lung is subjected to perfusion by the lung perfusion fluid provided by the invention; and finally, the preparation method for the lung perfusion fluid is simple, and raw materials are readily available and are economical and practical, so that the lung perfusion fluid is suitable for being popularized and applied to general laboratories or hospitals.
Description
Technical field
The present invention relates to medical biotechnology field, particularly relate to a kind of lung perfusate for donor lung perfusion
And preparation method thereof.
Background technology
The end that lung transplantation (Lung Transplantation, LTx) has become as medicine or surgical operation cannot be treated
Latter stage lung disease standard treatment regimen.Current standard compliant donor lung number cannot meet transplanting need
Ask, cause some patients dead during waiting for lung.Can improve in the method for lung shortage present situation at present,
Isolated lung perfusion (Ex Vivo Lung Perfusion, EVLP) is to be applied to lung transplantation field one extend for lung
Holding time and reparation supply the technology of injury of lung.EVLP can be with immediate assessment donor lung function, for medicine even
Gene therapy provides platform, improves " edge is for lung " (Marginal Donor Lungs) function, adds and meet
Transplanting condition transplants the quantity of lung, alleviates the present situation lacked for lung to a certain extent.Steen etc. with based on
The perfusate of perfadex liquid is applied to improve " edge is for lung " function, and 3 examples " edge is for lung " are filled by EVLP
Note (61～121min) successful implantation afterwards.Owing to the perfusate formula of external high-quality maintains secrecy, domestic lung perfusion
The formula of liquid and configuration technique can not meet the needs of donor lung perfusion, and lung preversation and Effect of Resuscitation be not strong,
The short problem being usually present of Hemoperfusion time.
Summary of the invention
The present invention solves that a kind of preparation that the problems referred to above of the prior art propose is simple, lung preversation is with multiple simultaneously
Su Zuoyong, and the lung perfusate for donor lung perfusion of Hemoperfusion time length and preparation method thereof.
In order to realize above-mentioned technical purpose, the technical measures that the present invention is taked are:
A kind of lung perfusate for donor lung perfusion, by purified water, dextran 20, sodium chloride, chlorine
Change potassium, magnesium sulfate, disodium hydrogen phosphate,anhydrous, potassium dihydrogen phosphate, glucose, calcium chloride, albumin, nitric acid
Glycerol, prednisolone, heparin sodium, cefazolin sodium and buffer agent configuration form.
In order to optimize technique scheme further, the technical measures that the present invention is taked also include:
Preferably, the lung perfusate of the present invention in preparation, using 1L purified water as solvent in terms of, be contained within
40-60g dextran 20,6-10g sodium chloride, 0.2-0.6g potassium chloride, 0.09-0.1g magnesium sulfate, 0.04-0.05g
Disodium hydrogen phosphate,anhydrous, 0.06-0.07g potassium dihydrogen phosphate, 0.8-0.9g glucose, 0.05-0.07g calcium chloride,
30-60g albumin, 1-4mg nitroglycerin, 200-300mg prednisolone, 4000-6000u heparin sodium and 0.4-0.6g
Cefazolin sodium, and use Tris-HCl buffer agent to regulate pH to 7.4-7.6..
It is further preferred that in the preparation of above-mentioned lung perfusate, using 1L purified water as solvent in terms of, be contained within 50g
Dextran 20,8g sodium chloride, 0.4g potassium chloride, 0.098g magnesium sulfate, 0.046g disodium hydrogen phosphate,anhydrous,
0.063g potassium dihydrogen phosphate, 0.9g glucose, 0.06g calcium chloride, 50g albumin, 2mg nitroglycerin,
250mg prednisolone, 5000u heparin sodium and 0.5g cefazolin sodium, use Tris-HCl buffer agent regulation pH
To 7.5.
On the other hand, the present invention also provides for a kind of as claimed in claim 1 for the lung filling of donor lung perfusion
The preparation method of flow liquid, comprises the following steps:
Step 1, the preparation of basic liquid
Take dextran heating to be dissolved in purified water, be sequentially added into sodium chloride, potassium chloride, magnesium sulfate, anhydrous phosphorus
Acid disodium hydrogen, potassium dihydrogen phosphate and glucose, use Tris-HCl buffer agent to adjust pH to 7.4-7.6, uses thin
Bacterium filter filters, and fills into purified water constant volume, obtains basic liquid；
Step 2, the preparation of pre-perfusate
The basic liquid obtained in step 1 is sequentially added into calcium chloride, albumin and nitroglycerin, uses
PH to 7.4-7.6 adjusted by Tris-HCl buffer agent, uses bacterial filter to filter, obtains pre-perfusate；
Step 3, prepares lung perfusate
The pre-perfusate obtained in step 2 adds prednisolone, heparin sodium and cefazolin sodium and obtains for transplanting
Lung perfusate for lung perfusion.
In above-mentioned preparation method, in step 1, the preparation of basic liquid is preferably: for taking 50g Dextran-20
Heating is dissolved in purified water, is sequentially added into 8g sodium chloride, 0.4g potassium chloride, 0.098g magnesium sulfate, 0.046g
Disodium hydrogen phosphate,anhydrous, 0.063g potassium dihydrogen phosphate and 0.9g glucose, use Tris-HCl buffer agent to adjust pH
To 7.4-7.6, use bacterial filter to filter, fill into purified water and be settled to 1000ml, obtain basic liquid.
In above-mentioned preparation method, being formulated as in described step 1 of pre-perfusate in step 2 in step 2
To basic liquid in be sequentially added into 0.06g calcium chloride, 50g albumin and 2mg nitroglycerin, use Tris-HCl
PH to 7.4-7.6 adjusted by buffer agent, uses bacterial filter to filter, obtains pre-perfusate.
In above-mentioned preparation method, preparing lung perfusate in step 3 is the pre-perfusion obtained in described step 2
Liquid adds 250mg prednisolone, 5000u heparin sodium and 0.5g cefazolin sodium and obtains for donor lung perfusion
Lung perfusate.
Further, the aperture of above-mentioned bacterial filter is preferably 0.22um.
The present invention uses technique scheme, compared with prior art, has the following technical effect that
The lung perfusate for donor lung perfusion that the present invention provides, first can provide good lung preversation and
Effect of Resuscitation, it is possible to meet the perfusion of donor lung and preserve needs；Secondly Hemoperfusion time is longer, through this
The lung that supplies after 6 hours perfusions of bright lung perfusate remains able to possess good lung compliance and oxygenate ability；?
After, the preparation method of the present invention is simple, and raw material is readily available and economical and practical, is suitable at general laboratory
Or clinical application application.
Detailed description of the invention
The invention provides a kind of lung perfusate for donor lung perfusion, by purified water, dextran, chlorine
Change sodium, potassium chloride, magnesium sulfate, disodium hydrogen phosphate,anhydrous, potassium dihydrogen phosphate, glucose, calcium chloride, white egg
In vain, nitroglycerin, prednisolone, heparin sodium, cefazolin sodium and buffer agent configuration form.
Below by specific embodiment, the present invention is carried out detailed and concrete introduction, so that being better understood from this
Bright, but following embodiment is not limiting as the scope of the invention.
Specific embodiment includes preparing embodiment and checking embodiment, the experimental apparatus of employing and the following institute of reagent
Show:
Water purification machine (Millipore, Germany)；Electronic analytical balance (Sartorius, Germany)；Cardiopulmonary bypass unit (Jostra
H20, Maquet, Germany)；Anesthetic machine (Detax Ohmeda, the U.S.)；Respirator (Detax Ohmeda,
Generalelectric, the U.S.) vacuum pump (SHZ-D (III) type, Shanghai)；Disposable sterilized filter (MF75,
U.S. nalgene).
Dextran-20, sodium chloride, potassium chloride, magnesium sulfate, disodium hydrogen phosphate (anhydrous), potassium dihydrogen phosphate,
Glucose, (purchased from Shanghai Ling Feng chemical reagent company limited).
Tris-HCl buffer agent (purchased from sigma company), bovine serum albumin (Albumin from bovine
Serum, BSA, sigma).
Nitroglycerin injection (Beijing Yimin Pharmaceutical Co., Ltd.), Urbason Solubile (Pfizer),
Heparin sodium injection (Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd), cefazolin sodium for injection (Zhuhai federalism
Medicine limited company).
The preparation of embodiment one lung perfusate
Use following step preparation for the lung perfusate of donor lung perfusion
Step 1, the preparation of basic liquid
Take the heating of 50g dextran to be dissolved in purified water, be sequentially added into 8g sodium chloride, 0.4g potassium chloride, 0.098g
Magnesium sulfate, 0.046g disodium hydrogen phosphate,anhydrous, 0.063g potassium dihydrogen phosphate and 0.9g glucose, use Tris-HCl
PH to 7.5 adjusted by buffer agent, and the bacterial filter using aperture to be 0.22um filters, and fills into purified water and is settled to
1000ml, obtains basic liquid；
Step 2, the preparation of pre-perfusate
The basic liquid obtained in step 1 is sequentially added into 0.06g calcium chloride, 50g albumin and 2mg nitric acid
Glycerol, uses Tris-HCl buffer agent to adjust pH to 7.4-7.6, the bacterial filter mistake using aperture to be 0.22um
Filter, obtains pre-perfusate；
Step 3, prepares lung perfusate
The pre-perfusate obtained in step 2 adds 250mg prednisolone, 5000u heparin sodium and 0.5g cephalo
Oxazoline sodium obtains the lung perfusate for donor lung perfusion.
The Pulmonis Sus domestica perfusion experiment of the lung perfusate of embodiment two present invention
Obtain pig supplies lung, and tracheal intubation is also connected to respirator.Pulmonary artery and left auricle direct cannulation also connect
To perfusion loop.Two intrusion pipes are first set level, and pour out the liquid of the inside, then see whether pressure-measuring pipe is full of liquid
Body, can flow in thick intrusion pipe, now open three junction blocks identical with air, be initially added into after zeroing again
Perfusate, starts perfusion, and perfusion uses the lung perfusate that embodiment one prepares.
Just start the cycle over temperature and be 12-13 DEG C, when the temperature of infusion liquid reaches 32 DEG C, carry out breathing machine ventilation,
Oxygen concentration is 0.21%, with 7ml tidal volume/kilogram beginning, 7 breaths/min and 5cmH2O PEEP frequency
Rate, the deoxidation of the infusion liquid of lung will be with nitrogen (86%), carbon dioxide (8%), and the admixture of gas of oxygen
Middle beginning (6%).I:E (inhaling/exhale ratio)=1:2.Do vim and vigour and a pulmonary ventilation per hour, interval
30 minutes.Do vim and vigour first 5 minutes: i.e. 55 timesharing and respirator be set to tidal volume TV 7 → 10ml/kg,
PEEP 5, RR (respiratory frequency) 7 → 10, oxygen concentration from 21% liter gradually to 100%.Finish handle after vim and vigour
Oxygen concentration recalls to 21%.Measure pulmonary artery partial pressure of oxygen, left room partial pressure of oxygen, calculate lung Arteriovenous Difference in Oxygen,
Measure pulmonary artery pressure and PLA left atrial pressure, calculate pulmonary vascular resistance.
In the present embodiment, utilize the lung perfusate perfusion pig of the present invention in vitro for lung, use 3 different pigs from
Body has carried out 3 times repeating test altogether for lung, all uses such as the lung perfusate of embodiment one preparation, carries out the most simultaneously
Pulmonary vascular resistance and pulmonary arterio venous oxygen partial pressure difference measure, and result is as shown in the table:
In 1 three perfusion experiments of table, pulmonary vascular resistance and pulmonary arterio venous oxygen partial pressure difference measure
As can be seen from the above table, using the lung perfusate of the present invention to carry out perfusion, not only Hemoperfusion time can maintain
Longer, pulmonary vascular resistance and lung Arteriovenous Difference in Oxygen all can produce a desired effect simultaneously.
The lung perfusate for donor lung perfusion that the present invention provides, first can provide good lung preversation and
Effect of Resuscitation, it is possible to meet the perfusion of donor lung and preserve needs；Secondly Hemoperfusion time is longer, through this
The lung that supplies after 6 hours perfusions of bright lung perfusate remains able to possess good lung compliance and oxygenate ability；?
After, the preparation method of the present invention is simple, and raw material is readily available and economical and practical, is suitable at general laboratory
Or clinical application application.
Being described in detail the specific embodiment of the present invention above, but it is intended only as example, the present invention is also
It is not restricted to particular embodiments described above.To those skilled in the art, any the present invention is carried out
Equivalent modifications and substitute the most all among scope of the invention.Therefore, without departing from the spirit of the present invention and model
Enclose lower made impartial conversion and amendment, all should contain within the scope of the invention.
Claims (8)
1. the lung perfusate for donor lung perfusion, it is characterised in that described lung perfusate by purified water,
Dextran 20, sodium chloride, potassium chloride, magnesium sulfate, disodium hydrogen phosphate,anhydrous, potassium dihydrogen phosphate, Fructus Vitis viniferae
Sugar, calcium chloride, albumin, nitroglycerin, prednisolone, heparin sodium, cefazolin sodium and buffer agent configuration and
Become.
A kind of lung perfusate for donor lung perfusion the most according to claim 1, it is characterised in that institute
State in the preparation of lung perfusate, using 1L purified water as solvent in terms of, be contained within 40-60g dextran 20,
6-10g sodium chloride, 0.2-0.6g potassium chloride, 0.09-0.1g magnesium sulfate, 0.04-0.05g anhydrous phosphoric acid hydrogen two
Sodium, 0.06-0.07g potassium dihydrogen phosphate, 0.8-0.9g glucose, 0.05-0.07g calcium chloride, 30-60g are white
Albumen, 1-4mg nitroglycerin, 200-300mg prednisolone, 4000-6000u heparin sodium and 0.4-0.6g
Cefazolin sodium, and use Tris-HCl buffer agent to regulate pH to 7.4-7.6.
A kind of lung perfusate for donor lung perfusion the most according to claim 1, it is characterised in that institute
State in the preparation of lung perfusate, using 1L purified water as solvent in terms of, be contained within 50g dextran 20,8g
Sodium chloride, 0.4g potassium chloride, 0.098g magnesium sulfate, 0.046g disodium hydrogen phosphate,anhydrous, 0.063g di(2-ethylhexyl)phosphate
Hydrogen potassium, 0.9g glucose, 0.06g calcium chloride, 50g albumin, 2mg nitroglycerin, 250mg first are strong
Dragon, 5000u heparin sodium and 0.5g cefazolin sodium, use Tris-HCl buffer agent regulation pH to 7.5.
4. the preparation method of the lung perfusate for donor lung perfusion as claimed in claim 1, it is characterised in that
Comprise the following steps:
Step 1, the preparation of basic liquid
Take dextran heating to be dissolved in purified water, be sequentially added into sodium chloride, potassium chloride, magnesium sulfate, anhydrous phosphoric acid hydrogen
Disodium, potassium dihydrogen phosphate and glucose, use Tris-HCl buffer agent to adjust pH to 7.4-7.6, use antibacterial
Filter filters, and fills into purified water constant volume, obtains basic liquid；
Step 2, the preparation of pre-perfusate
The basic liquid obtained in step 1 is sequentially added into calcium chloride, albumin and nitroglycerin, uses Tris-HCl
PH to 7.4-7.6 adjusted by buffer agent, uses bacterial filter to filter, obtains pre-perfusate；
Step 3, prepares lung perfusate
The pre-perfusate obtained in step 2 adds prednisolone, heparin sodium and cefazolin sodium and obtains for donor lung
The lung perfusate of perfusion.
Preparation method the most according to claim 4, it is characterised in that the preparation of basic liquid in described step 1
For take 50g Dextran-20 heating be dissolved in purified water, be sequentially added into 8g sodium chloride, 0.4g potassium chloride,
0.098g magnesium sulfate, 0.046g disodium hydrogen phosphate,anhydrous, 0.063g potassium dihydrogen phosphate and 0.9g glucose,
Use Tris-HCl buffer agent to adjust pH to 7.4-7.6, use bacterial filter to filter, fill into purified water and be settled to
1000ml, obtains basic liquid.
Preparation method the most according to claim 4, it is characterised in that in described step 2, pre-perfusate joins
It is made as in the basic liquid obtained in described step 1 being sequentially added into 0.06g calcium chloride, 50g albumin and 2mg
Nitroglycerin, uses Tris-HCl buffer agent to adjust pH to 7.4-7.6, uses bacterial filter to filter, obtains pre-
Perfusate.
Preparation method the most according to claim 4, it is characterised in that prepare lung perfusate in described step 3
Pre-perfusate for obtaining in described step 2 adds 250mg prednisolone, 5000u heparin sodium and 0.5g head
Spore oxazoline sodium obtains the lung perfusate for donor lung perfusion.
Preparation method the most according to claim 4, it is characterised in that the aperture of described bacterial filter is
0.22um。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110267534A (en) * | 2017-01-17 | 2019-09-20 | 西维沃医疗科技有限公司 | Organ saves and/or primer solution |
| CN112273373A (en) * | 2020-10-29 | 2021-01-29 | 无锡市人民医院 | Isolated lung mechanical perfusion liquid and preparation method and application thereof |
| CN112425602A (en) * | 2019-08-26 | 2021-03-02 | 无锡市人民医院 | Isolated lung mechanical perfusion liquid and preparation method and application thereof |
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| CN103131666A (en) * | 2011-11-23 | 2013-06-05 | 四川农业大学 | Goose primary hepatocyte isolation culture method |
| CN103841824A (en) * | 2011-04-14 | 2014-06-04 | 特兰斯迈迪茨公司 | Organ care solution for ex-vivo machine perfusion of donor lungs |
| CN104996396A (en) * | 2015-07-27 | 2015-10-28 | 中山大学附属第一医院 | Perfusion preservation fluid for isolated liver |
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2016
- 2016-04-28 CN CN201610277886.3A patent/CN105918308A/en active Pending
Patent Citations (4)
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| WO1997043899A1 (en) * | 1996-05-17 | 1997-11-27 | Breonics, Inc. | Solution and process for resuscitation and reparation of ischemically damaged tissue |
| CN103841824A (en) * | 2011-04-14 | 2014-06-04 | 特兰斯迈迪茨公司 | Organ care solution for ex-vivo machine perfusion of donor lungs |
| CN103131666A (en) * | 2011-11-23 | 2013-06-05 | 四川农业大学 | Goose primary hepatocyte isolation culture method |
| CN104996396A (en) * | 2015-07-27 | 2015-10-28 | 中山大学附属第一医院 | Perfusion preservation fluid for isolated liver |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110267534A (en) * | 2017-01-17 | 2019-09-20 | 西维沃医疗科技有限公司 | Organ saves and/or primer solution |
| CN110267534B (en) * | 2017-01-17 | 2021-11-16 | 西维沃医疗科技有限公司 | Organ preservation and/or perfusion solution |
| CN112425602A (en) * | 2019-08-26 | 2021-03-02 | 无锡市人民医院 | Isolated lung mechanical perfusion liquid and preparation method and application thereof |
| CN112273373A (en) * | 2020-10-29 | 2021-01-29 | 无锡市人民医院 | Isolated lung mechanical perfusion liquid and preparation method and application thereof |
| CN112273373B (en) * | 2020-10-29 | 2022-06-07 | 无锡市人民医院 | Isolated lung mechanical perfusion liquid and preparation method and application thereof |
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Application publication date: 20160907 |