CN102550541B - Organ vitrificated cryopreserration liquid - Google Patents
Organ vitrificated cryopreserration liquid Download PDFInfo
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- CN102550541B CN102550541B CN201010601358.1A CN201010601358A CN102550541B CN 102550541 B CN102550541 B CN 102550541B CN 201010601358 A CN201010601358 A CN 201010601358A CN 102550541 B CN102550541 B CN 102550541B
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Abstract
The present invention relates to medical organ and preserve field, specifically disclose the vitrificated cryopreserration liquid preserved for a long time for organ, it is characterized in that this conserving liquid is made up of compositions such as dimethyl sulfoxide (DMSO), acetamide, glycerine, ethylene glycol, propane diols, sucrose, polyethylene glycol, dimethyl sulfoxide (DMSO), acetamide, glycerine, ethylene glycol, propane diols, sucrose concentration are 0.1-6mo/L, Polyethylene glycol is 1-20mmol/L, and pH value scope is 5.0-8.0.Implement according to the glassy state storage liquid preserved for organ provided by the invention; ice crystal is not formed in glassy state in cooling refrigerating process; protective tissue Morphology and structure integrality; under freezing conditions suppress histiocytic energy metabolism completely; its biological function is recovered after recovery; can be used for the freezen protective of kidney, also can further genralrlization be applied to other organs long-term frozen preserve, for scientific research and clinical practice.
Description
Technical field
Kidney is the organ of function complexity in human body, can only maximum 50 hours of Plantlet in vitro under 4 DEG C of perfusion conditions.Want long-term even persistence kidney, storage temperature must be reduced further, suppress histiocytic metabolic rate, even make cellular metabolism stop completely.Kidney if preserve at suitable long period under refrigeration, when needed by freezen protective adopt suitable means " recovery " for kidney after transplant again, a brand-new prospect will be opened up for clinical transplantation work.
Freezing and storing method is considered to realize the desirable route that organ is preserved for a long time, the key of problem is that organ mass is larger, in cooling and rewarming process, be difficult to overcome intraor extracellular ice crystal formed, the damage of machinery, chemistry, osmotic pressure and biology aspect can not be avoided, so the frozen problem of larger volume organ is still unresolved so far simultaneously.Vitrifying (Vitrification) refers to that liquid rotating becomes the solidification process of amorphous state (glassy state).Relation between glassy solids molecule and liquid without obviously difference, does not form ice crystal in cooling refrigerating process, because of but a kind of desirable freezen protective approach.
Background technology
In the recent period, Cell Cryopreservation, sperm, embryo even ovary tissue successively obtain successfully, but complex organ is as the freezen protective of kidney, are but still difficult problems not yet broken through.The world today, along with the continuous increase with nephrotic's quantity that develops rapidly of kidney transplant technology, stocks in the urgent need to a large amount of kidneys and supplies.Current clinical transplantation is all preserved for kidney more than 0 DEG C, although the method comparative maturity, easy, the holding time is very limited.Although Chinese scholars sets about extending the kidney being kept in existence time from side such as improvement conserving liquid level and conserving appliance etc., or within the limited holding time, improve tissue matching's work, to improve for the reasonable employment rate of kidney and the long-term surviving rate after transplanting, but still clinical demand can not be met.In addition, when donor organ lacks at home, certain organ wasting of resources can also be caused because distribution type, patient's condition are not inconsistent with reasons such as regions.The kidney that human leucocyte antigen (HLA) (HLA) distribution type is coincide can be shared within the scope of a country and even All Around The World, each receptor can be obtained in time optimized for kidney? for Utopian like this hope, only have by improving kidney being kept in existence technology, extend for even several years kidney holding time to several months, and set up " organ storehouse " (OrganBank) and could finally realize.
Summary of the invention
Organ vitrificated cryopreserration liquid of the present invention, be made up of compositions such as dimethyl sulfoxide (DMSO), acetamide, glycerine, ethylene glycol, propane diols, sucrose, polyethylene glycol, dimethyl sulfoxide (DMSO), acetamide, glycerine, ethylene glycol, propane diols, sucrose concentration are 0.1-6mol/L, Polyethylene glycol is 1-20mmol/L, and pH value scope is 5.0-8.0.Vitrificated cryopreserration liquid solution in cooling refrigerating process be glassy state, does not form ice crystal, avoids or reduce the freezing infringement of cell, can the integrality of Cell protection Structure and form, while suppress histiocytic energy metabolism completely.After the recovery of suitable means, metabolic activity in cells is restored, and again can play biological function.Organ vitrificated cryopreserration liquid of the present invention, can be used for the freezen protective (-200--30 DEG C) of kidney, and the long-term frozen that also can be applicable to other organs (liver, heart etc.) is preserved.
Embodiment
Through organ vitrificated cryopreserration perfusion kidney of the present invention ,-80 DEG C will be cooled to, proceed to profound hypothermia refrigerator or liquid nitrogen is preserved for a long time.4 DEG C are warming up to, the laggard line correlation morphology of wash-out conserving liquid and function assessment inspection during organ recovery.
Preserved the new zealand white rabbit kidney of 2 weeks at-80 DEG C by organ vitrificated cryopreserration liquid of the present invention, general form has no obvious freezing infringement, skin medullary substance clear in structure, under light microscope, in glomerulus, Capillary loops is complete, the slight swelling of renal cells, glomerular filtration membrane under electron microscope, the ultra microstructure such as sertoli cell and mesangial cell is clear, nuclei dyeing chromaticness is dense, kytoplasm inner cell organ number is normal, face, epithelial cell chamber brush border microvillus marshalling, vascular endothelial cell and elastic membrane complete, display kidney tissue morphology after freezen protective has no obvious destruction.
Preserved the new zealand white rabbit kidney of 2 weeks at-80 DEG C by organ vitrificated cryopreserration liquid of the present invention, phenolsulfonphthalein excretion test (PSP test), indicarminum measuring renal tubular function is carried out after ex vivo perfusion, PSP15 minute and 120 minutes excretion amounts, all close to normal value (25%; 55%), at once see blue urine and discharge after indicarminum perfusion, display kidney has excretion function through freezen protective metanephric tubule.
Preserved the beasle dog kidney of 2 weeks at-80 DEG C by organ vitrificated cryopreserration liquid of the present invention, autotransplantation of kidney is carried out CDFI for postoperative 1 month, is launched the inspection such as single photon emission tomoscan (ECT) and vein secretion radiography (IVP), color ultrasound is transplanted blood flow in kidney as seen and is passed through, IVP and ECT shows renal secretion/filtering function sustainable existence.
Claims (4)
1. an organ vitrificated cryopreserration liquid, it is characterized in that, be made up of dimethyl sulfoxide (DMSO), acetamide, glycerine, ethylene glycol, propane diols, sucrose, polyethylene glycol, dimethyl sulfoxide (DMSO), acetamide, glycerine, ethylene glycol, propane diols, sucrose concentration are 0.1-6mol/L, Polyethylene glycol is 1-20mmol/L, and pH value scope is 5.0-8.0.
2. conserving liquid according to claim 1 and 2, is characterized in that, described organ is kidney, liver, heart, lungs, skin or limbs.
3. an organ store method, comprises and organ is put into conserving liquid, it is characterized in that, described conserving liquid is organ vitrificated cryopreserration liquid according to claim 1, and storage temperature is-200--30 degree Celsius.
4. organ vitrificated cryopreserration liquid according to claim 1 is used for organ preservation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201010601358.1A CN102550541B (en) | 2010-12-23 | 2010-12-23 | Organ vitrificated cryopreserration liquid |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201010601358.1A CN102550541B (en) | 2010-12-23 | 2010-12-23 | Organ vitrificated cryopreserration liquid |
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| CN102550541A CN102550541A (en) | 2012-07-11 |
| CN102550541B true CN102550541B (en) | 2016-02-17 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP7061610B2 (en) | 2016-12-20 | 2022-04-28 | ティシュー テスティング テクノロジーズ エルエルシー | Ice-free storage of large tissue samples for practical and functional tissue banking |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104585165A (en) * | 2015-02-24 | 2015-05-06 | 陈德琼 | Human organ freezing protective solution |
| CN104904707A (en) * | 2015-05-29 | 2015-09-16 | 广州赛莱拉干细胞科技股份有限公司 | Composition, application of composition, vitrification cryopreserved agent of placenta and preparation method |
| CN105241726A (en) * | 2015-09-14 | 2016-01-13 | 黄小军 | Immunohistochemical staining method and applications of immunohistochemical staining method in cervical tumorous lesion screening |
| CN107236340B (en) * | 2017-05-22 | 2019-04-23 | 安徽宏远职业卫生技术服务有限公司 | A kind of cuvette store method |
| CN111657267B (en) * | 2020-06-17 | 2021-02-02 | 科瑞百奥泰州生物技术有限公司 | Ice-free crystal frozen preservation solution and freezing method for preservation of cartilage, tendon and meniscus |
| CN117717066A (en) * | 2024-02-04 | 2024-03-19 | 广东海洋大学 | Embryo preservation solution, cryopreservation method thereof and application of embryo preservation solution in cryopreservation of Babylonia embryos |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002032225A2 (en) * | 2000-10-19 | 2002-04-25 | Organ Recovery Systems | Method of cryopreservation of tissues or organs other than a blood vessel by vitrification |
| CN1986780A (en) * | 2006-12-14 | 2007-06-27 | 宁夏医学院 | New-type of vetrifying solution |
| CN101037667A (en) * | 2007-02-09 | 2007-09-19 | 浙江大学 | Ultra-low temperature preservation solution for animal cells and preserving method |
| CN101253856A (en) * | 2008-04-11 | 2008-09-03 | 中山大学 | Vitrification frozen stock solution |
| WO2010096821A2 (en) * | 2009-02-23 | 2010-08-26 | Cell & Tissue Systems, Inc. | Method for ice-free cryopreservation of tissue |
-
2010
- 2010-12-23 CN CN201010601358.1A patent/CN102550541B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002032225A2 (en) * | 2000-10-19 | 2002-04-25 | Organ Recovery Systems | Method of cryopreservation of tissues or organs other than a blood vessel by vitrification |
| CN1986780A (en) * | 2006-12-14 | 2007-06-27 | 宁夏医学院 | New-type of vetrifying solution |
| CN101037667A (en) * | 2007-02-09 | 2007-09-19 | 浙江大学 | Ultra-low temperature preservation solution for animal cells and preserving method |
| CN101253856A (en) * | 2008-04-11 | 2008-09-03 | 中山大学 | Vitrification frozen stock solution |
| WO2010096821A2 (en) * | 2009-02-23 | 2010-08-26 | Cell & Tissue Systems, Inc. | Method for ice-free cryopreservation of tissue |
Non-Patent Citations (1)
| Title |
|---|
| 动物胚胎的玻璃化冻存;胡军祥 等;《科技通报》;20051130;第21卷(第6期);第680页左栏 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP7061610B2 (en) | 2016-12-20 | 2022-04-28 | ティシュー テスティング テクノロジーズ エルエルシー | Ice-free storage of large tissue samples for practical and functional tissue banking |
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| CN102550541A (en) | 2012-07-11 |
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