CN102144629A - Organ cryopreservation fluid - Google Patents

Organ cryopreservation fluid Download PDF

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Publication number
CN102144629A
CN102144629A CN2010190910082A CN201019091008A CN102144629A CN 102144629 A CN102144629 A CN 102144629A CN 2010190910082 A CN2010190910082 A CN 2010190910082A CN 201019091008 A CN201019091008 A CN 201019091008A CN 102144629 A CN102144629 A CN 102144629A
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CN
China
Prior art keywords
organ
cryopreservation
kidney
acetamide
glucose
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Pending
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CN2010190910082A
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Chinese (zh)
Inventor
肖传国
潘峰
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XIAO ZHUANGUO
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XIAO ZHUANGUO
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Application filed by XIAO ZHUANGUO filed Critical XIAO ZHUANGUO
Priority to CN2010190910082A priority Critical patent/CN102144629A/en
Publication of CN102144629A publication Critical patent/CN102144629A/en
Pending legal-status Critical Current

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Abstract

The invention relates to the medical organ preservation field, and in particular discloses an organ cryopreservation fluid for preserving the organ for a long time. The organ cryopreservation fluid is characterized by comprising the ingredients such as dimethyl sulfoxide, acetamide, glucose, glycerol, polyethylene glycol and the like; the concentrations of the dimethyl sulfoxide, the acetamide, the glucose and the glycerol are 0.2 to 5 mol/L; and the concentration of the polyethylene glycol is 1 to 20 mol/L. The cryopreservation fluid for preserving the organs in the invention can protect shape and structure integrity of tissue cells under the cryopreservation under the condition that energy metabolism is fully inhibited, and recover biological function after proper condition anabiosis; and the cryopreservation fluid can be used for cryopreservation for kidneys, and also can be applied to cryopreservation for other organs so that the organs can be preserved for a long time in clinical practices.

Description

The organ profound hypothermia is preserved liquid
Technical field
The profound hypothermia store method is considered to realize the desirable approach of organ long preservation, the key of problem is that the organ volume is bigger, ice crystal forms be difficult to overcome cell in cooling and rewarming process inside and outside, can not avoid simultaneously the damage of machinery, chemistry, osmotic pressure and biology aspect, so the frozen problem of larger volume organ is still unresolved so far.Kidney is the organ of function complexity in the human body, can only {in vitro} conservation under 4 ℃ of perfusion conditions maximum 50 hours.For kidney if can be under suitable profound hypothermia condition long preservation, will open up a brand-new prospect for clinical transplantation work.
Background technology
In the recent period, profound hypothermia is preserved cell, sperm, embryo even ovary tissue and is successively achieved success, but for example profound hypothermia preservation of kidney of complex organ but still is a difficult problem that does not break through as yet.In the world today, nephrotic's quantity constantly increases, and the technology of kidney transplant is also developing rapidly, and this stocks with regard to a large amount of kidney of needs and supplies.What present clinical kidney transplant was used is all preserving more than 0 ℃ for kidney, though the method comparative maturity, easy, but the holding time is limited, Chinese scholars sets about prolonging the time that kidney is preserved from improving aspects such as preserving liquid level and conserving appliance, or in the limited holding time, improve tissue matching and work, supply the reasonable utilization rate of kidney and the long-term surviving rate after the transplanting with raising, but still can not satisfy clinical demand.In addition, at home under the situation that donor organ lacks, owing to join type, patient's condition is not inconsistent and reasons such as region also can cause certain organ wasting of resources.Can in a country and even All Around The World scope, share human leucocyte antigen (HLA) (HLA) and join the kidney that type coincide, is it optimized for kidney that each receptor can both in time be obtained? for Utopian like this hope, have only by improving kidney preservation technology, prolong for kidney holding time to several weeks even several years, and set up " organ storehouse " (Organ Bank) and could finally realize.But, preserve for a long time for kidney, must further reduce the temperature that kidney is preserved, suppress histiocytic metabolic rate, even cellular metabolism is stopped fully.That will preserve when needed then, adopts suitable means to transplant after " recovery " for kidney again.
Summary of the invention
Organ profound hypothermia of the present invention is preserved liquid, form by compositions such as dimethyl sulfoxide (DMSO), acetamide, glucose, glycerine, polyethylene glycol, dimethyl sulfoxide (DMSO), acetamide, glucose, glycerol concentration are 0.2~5mol/L, and polyethylene glycol concentration is 1~20mmol/L.Under the profound hypothermia condition that the energy metabolism of organ biological cell is suppressed fully; this preservation liquid can be protected the cellular morphology structural intergrity; make it to be in resting state; avoid or reduce the freezing infringement of profound hypothermia pair cell; after suitable condition recovery; metabolic activity in cells is recovered, brought into play its biological function.Organ profound hypothermia of the present invention is preserved liquid, and the profound hypothermia that can be used for kidney is preserved (200 ℃~-20 ℃), and the profound hypothermia that also can be applicable to other organs is preserved, and realizes the target of organ long preservation.
Embodiment
To put into through the kidney after organ profound hypothermia of the present invention is preserved perfusion and fill the freezing storing box of preserving liquid, service routine frigorimeter control speed is freezing, after reducing to-30 ℃ or-80 ℃ and keeping 1 hour, changes-30 ℃ or-80 ℃ of refrigerators over to.Preserve after 30 days service routine frigorimeter control speed and be warming up to 4 ℃, wash-out is preserved liquid and is carried out linked groups's pathological examination and other operations.
Preserve liquid by organ profound hypothermia of the present invention and carry out new zealand white rabbit kidney after profound hypothermia is preserved, general form is not seen obvious freezing infringement, skin medullary substance clear in structure, the interior capillary button loop of glomerulus is high-visible under the light microscope, Bowman's capsule does not have expansion, the slight cloudy swelling of renal cells, glomerular filtration membrane under the electron microscope, ultra microstructure such as sertoli cell and mesangial cell is complete, the nuclei dyeing chromaticness is dense, kytoplasm inner cell organ number is normal, epithelial cell chamber face brush border microvillus marshalling, vascular endothelial cell and elastic membrane are complete, show that kidney tissue morphology after profound hypothermia is preserved is complete.
Preserve liquid by organ profound hypothermia of the present invention and carry out profound hypothermia preservation back new zealand white rabbit kidney, carry out phenolsulfonphthalein excretion test (PSP test), indicarminum measuring renal tubular function behind the ex vivo perfusion, the excretion amount was 28.5% in PSP15 minute, the excretion amount was 57.6% in 120 minutes, and all the excretion amount is higher than normal value (25%; 55%), indicarminum pours into the back can see blue urine discharge a moment, shows that kidney is normal through profound hypothermia preservation metanephric tubule excretion function.
Preserve liquid by organ profound hypothermia of the present invention and carry out beasle dog kidney after profound hypothermia is preserved, the autotransplantation of kidney postoperative carries out CDFI, emission single photon computed tomography (ECT) and the inspections such as (IVP) of vein secretion radiography, as seen postoperative check color ultrasound transplants that blood flow passes through in the kidney, and IVP and ECT show that also renal function continues to exist.

Claims (6)

1. an organ profound hypothermia is preserved liquid, it is characterized in that, comprises compositions such as dimethyl sulfoxide (DMSO), acetamide, glucose, glycerine, polyethylene glycol.
2. preservation liquid according to claim 1 is characterized in that dimethyl sulfoxide (DMSO), acetamide, glucose, glycerol concentration are 0.2~5mol/L, and polyethylene glycol concentration is 1~20mmol/L.
3. require 1 described preservation liquid according to economic rights, it is characterized in that, profound hypothermia is-200~-20 degrees centigrade.
4. according to the arbitrary described preservation liquid of claim 1 to 3, it is characterized in that described organ is kidney, liver, heart, lungs, skin or limbs etc.
5. an organ store method comprises organ is put into preservation liquid, it is characterized in that, comprises compositions such as dimethyl sulfoxide (DMSO), acetamide, glucose, glycerine, polyethylene glycol in the described preservation liquid.
6. store method according to claim 5 is characterized in that, organ is carried out profound hypothermia preserve.
CN2010190910082A 2010-02-08 2010-02-08 Organ cryopreservation fluid Pending CN102144629A (en)

Priority Applications (1)

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Application Number Priority Date Filing Date Title
CN2010190910082A CN102144629A (en) 2010-02-08 2010-02-08 Organ cryopreservation fluid

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102487938A (en) * 2011-11-15 2012-06-13 青岛大学医学院附属医院 Cryopreservation method for protecting cell junction
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
CN106047812A (en) * 2016-05-25 2016-10-26 上海赛立维生物科技有限公司 Tumor tissue cryopreservation and resuscitation kit and treatment method adopted by same
CN106857501A (en) * 2017-02-22 2017-06-20 单纯 Storing liquid and preparation method thereof for preserving rat liver homogenate S9
CN107560918A (en) * 2017-09-22 2018-01-09 漯河医学高等专科学校 A kind of pathological tissue specimen fixer

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102487938A (en) * 2011-11-15 2012-06-13 青岛大学医学院附属医院 Cryopreservation method for protecting cell junction
CN102487938B (en) * 2011-11-15 2014-04-02 青岛大学医学院附属医院 Cryopreservation method for protecting cell junction
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
CN106047812A (en) * 2016-05-25 2016-10-26 上海赛立维生物科技有限公司 Tumor tissue cryopreservation and resuscitation kit and treatment method adopted by same
CN106857501A (en) * 2017-02-22 2017-06-20 单纯 Storing liquid and preparation method thereof for preserving rat liver homogenate S9
CN106857501B (en) * 2017-02-22 2021-09-24 单纯 Storage liquid for storing rat liver homogenate S9 and preparation method thereof
CN107560918A (en) * 2017-09-22 2018-01-09 漯河医学高等专科学校 A kind of pathological tissue specimen fixer

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Application publication date: 20110810