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1. WO2020107911 - CRYOPRESERVATION AND RESUSCITATION METHOD FOR THREE-DIMENSIONAL RETINA TISSUE

Publication Number WO/2020/107911
Publication Date 04.06.2020
International Application No. PCT/CN2019/097717
International Filing Date 25.07.2019
IPC
C12N 5/071 2010.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
07Animal cells or tissues
071Vertebrate cells or tissues, e.g. human cells or tissues
A01N 1/02 2006.01
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
NPRESERVATION 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
1Preservation of bodies of humans or animals, or parts thereof
02Preservation of living parts
CPC
A01N 1/0221
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES
1Preservation of bodies of humans or animals, or parts thereof
02Preservation of living parts
0205Chemical aspects
021Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
0221Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
A01N 1/0226
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES
1Preservation of bodies of humans or animals, or parts thereof
02Preservation of living parts
0205Chemical aspects
021Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
0226Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
C12N 5/0621
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor;
06Animal cells or tissues; Human cells or tissues
0602Vertebrate cells
0618Cells of the nervous system
0621Eye cells, e.g. cornea, iris pigmented cells
Applicants
  • 中山大学中山眼科中心 ZHONGSHAN OPHTHALMIC CENTER, SUN YAT-SEN UNIVERSITY [CN]/[CN]
Inventors
  • 葛坚 GE, Jian
  • 罗子明 LUO, Ziming
  • 冼碧琨 XIAN, Bikun
  • 李凯婧 LI, Kaijing
  • 叶美芳 YE, Meifang
Agents
  • 广州科粤专利商标代理有限公司 GUANGZHOU KEYUE I.P. LAW OFFICE
Priority Data
201811451304.430.11.2018CN
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) CRYOPRESERVATION AND RESUSCITATION METHOD FOR THREE-DIMENSIONAL RETINA TISSUE
(FR) PROCÉDÉ DE CRYOCONSERVATION ET DE RÉANIMATION POUR TISSU RÉTINIEN TRIDIMENSIONNEL
(ZH) 一种三维视网膜组织的冷冻保存及复苏方法
Abstract
(EN)
Disclosed are a cryopreservation and resuscitation method for a three-dimensional retina tissue. The method mainly comprises the cryopreservation steps of: selecting a three-dimensional retina tissue suitable for cryopreservation, suspending the tissue in an RDM culture solution, and then transferring same to a cryopreservation tube, adding 2× cryopreserved fluid according to an isovolumetric ratio, taking away a portion of supernatant after the three-dimensional retina tissue sinks to the bottom of the tube, sealing the cryopreservation tube and placing same in -80°C overnight so as to lower the temperature, and then transferring the cryopreservation tube to liquid nitrogen to be stored; finally, resuscitating the cryopreserved retina tissue and then placing same into an incubator to be subjected to suspension culture. The method is good in repeatability and high in success rate, and the survival rate of resuscitating is up to 80%-90%, making the three-dimensional retina tissue controllable in a growing process and reliable in storage and transportation, and it is guaranteed that the resuscitated tissue is complete in structure and good in activity.
(FR)
Procédé de cryoconservation et de réanimation pour tissu rétinien tridimensionnel. Le procédé comprend principalement les étapes de cryoconservation consistant à sélectionner un tissu rétinien tridimensionnel approprié pour la cryoconservation, à mettre en suspension le tissu dans une solution de culture RDM, puis à le transférer à un tube de cryoconservation, à ajouter un fluide cryoconservé 2× selon un rapport isovolumétrique, à retirer une partie du surnageant après que le tissu rétinien tridimensionnel ait coulé au fond du tube, à sceller le tube de cryoconservation et à le placer à -80 °C pendant une nuit de façon à abaisser la température, puis à transférer le tube de cryoconservation dans de l'azote liquide pour être stocké; enfin, un étape de réanimation du tissu rétinien cryoconservé puis son placement dans un incubateur pour le soumettre à une culture en suspension. Le procédé présente une bonne répétabilité et un taux de réussite élevé, le taux de survie de la réanimation s'élevant jusqu'à 80 % à 90 %, ce qui rend le tissu rétinien tridimensionnel contrôlable dans un processus de croissance, fiable pour le stockage et le transport, et permet de garantir que le tissu de réanimation est complet en structure et de bonne activité.
(ZH)
一种三维视网膜组织的冷冻保存及复苏方法。该方法的冻存步骤主要为:选取适合冻存的三维视网膜组织,悬浮于RDM培养液,然后转移至冻存管,按等体积比加入2×冷冻保存液,待三维视网膜组织沉至管底后吸走部分的上清液,密封冻存管,置于-80℃中过夜降温,再移入液氮中保存;最后将保存的视网膜组织解冻复苏即可置于培养箱进行悬浮培养。该方法可重复性好,成功率高,复苏成活率可高达80~90%,使得三维视网膜组织的发育进程可控、保存运输可靠,并保证复苏的组织结构完整、活性良好。
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