CN106577632A - Device for freeze preservation of human ovarian tissue under liquid nitrogen - Google Patents

Device for freeze preservation of human ovarian tissue under liquid nitrogen Download PDF

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Publication number
CN106577632A
CN106577632A CN201611103273.4A CN201611103273A CN106577632A CN 106577632 A CN106577632 A CN 106577632A CN 201611103273 A CN201611103273 A CN 201611103273A CN 106577632 A CN106577632 A CN 106577632A
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China
Prior art keywords
liquid nitrogen
human ovarian
closure
under liquid
organizes
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CN201611103273.4A
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CN106577632B (en
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饶金鹏
金敏
钱小红
邱枫
金帆
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Zhejiang University ZJU
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Zhejiang University ZJU
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION 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
    • A01N1/00Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02Preservation of living parts
    • A01N1/0236Mechanical aspects
    • A01N1/0263Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
    • A01N1/0268Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Hematology (AREA)
  • Mechanical Engineering (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

A device for freeze preservation of a human ovarian tissue under liquid nitrogen comprises a pipe body; one end of the pipe body is provided with an opening, the opening is equipped with a sealing cover, the sealing cover and the pipe body can be detachably connected, and a sealed storage space is enclosed by the sealing cover and the pipe body; the inner top surface of the sealing cover is provided with a forking device for forking the human ovarian tissue. The forking device comprises a plurality of fine needles which are arranged in parallel, and fixed ends of the fine needles are vertically fixed on the inner end surface of the sealing cover, free ends of the fine needles extend to the pipe body along the axial direction of the pipe body. The forking device comprises 3 fine needles, the 3 fixed ends of the 3 fine needles are in head-to-tail connection to form a regular triangle, and the circle center of the inner end face of the sealing cover is the center of the regular triangle. The device is simple in structure and easy to use; the forking device can avoid physical damage of blind clipping with forceps under liquid nitrogen on an ovarian follicle enriched cortical part in the ovarian tissue, and also avoids cross contamination with other iatrogenic substances in a liquid nitrogen tank.

Description

A kind of human ovarian organizes cryopreservation device under liquid nitrogen
Technical field
The present invention relates to human fertility tries hard to keep deposits technical field, more particularly to a kind of human ovarian organizes frozen under liquid nitrogen Device.
Background technology
Heavy dose of systemic chemotherapy or radiotherapy in cancer healing process can have a strong impact on the fertility of young woman, there is statistics The young patient that malignant tumor is suffered from display receives after Radiotherapy chemotherapy 1/3 will be infertile.At present, the side of tumor patient fertility is protected Method mainly has 3 kinds:Embryo cryopreservation, oocyte freezing and ovary tissue freezing.But embryo cryopreservation is unsuitable for preadolescence or does not have Have with occasionally without the women of tribute essence;Oocyte freezing then needs the longer induced ovulation cycle, there is delay treatment tumor most The risk on good opportunity;Ovary tissue freezing is drawn materials not by time restriction, in female fertility because can simultaneously preserve a large amount of follicles Preservation aspect has wide potential applicability in clinical practice.
At present ovary tissue is frozen there is no unified standard, conventional cryopreservation methods to have sequencing freezing and vitrification at a slow speed cold Freeze.But sequencing freezing needs the Slow-rate freezing instrument of purchasing expensive, freezing whole process to need 2-4 hours, and operation is relative complex, and vitrification Freezing, then can make ovary tissue be in glassy state, it is to avoid intracellular ice in the temperature-fall period being exceedingly fast and in the extremely short time Brilliant formation, with it is quick, efficient, easily and economically the characteristics of and be used widely.
Conventional Vitrification of Ovarian Tissue method has:Frozen tube method, microdroplet Vitrification, solid surface vitrification Method and metallic sieve method.
Different from embryo and oocyte, ovary tissue has, cortex follicles regions sensitivity many containing cell quantity, species Fragile the features such as.Therefore, select suitable cryopreservation device most important during glass freezing, cooling is improved as far as possible Speed, makes ovary tissue reach glassy state, while reduce the stimulation to sensitive cortex follicles regions damaging.Frozen tube method because In operation first by ovary tissue tubulation seals, liquid nitrogen is subsequently put into again, actual is a kind of mediate contact liquid nitrogen method, its drop Warm speed is far below contact method, causes Follicles quantity and proliferation activity after recovery to be significantly lower than remaining method.Microdroplet Vitrification, solid surface vitrification method are direct liquid nitrogen contact method, although be attained by preferable rate of temperature fall, but throwing To enter be both needed to after liquid nitrogen and under liquid nitrogen ovary tissue is fitted in cryovial with tweezers, because liquid nitrogen has volatility, tubulature operation Interference is big, and tweezers grips that dynamics is wayward under liquid nitrogen, and physical property extruding damage is easily caused to sensitive cortex follicle area Wound.Metallic sieve method is that first ovary tissue is placed on metal grill, is loaded into again in stored frozen bag after input liquid nitrogen, though The directly contact of tweezers and ovary tissue is so avoided, but stored frozen bag resists mechanical stretch ability not as freezing at -196 DEG C Pipe is deposited, it is easy to crack, and stored frozen bag shape do not fix, and is difficult frozen in order.
The content of the invention
For the deficiencies in the prior art, the present invention provides a kind of effect that can reach very fast cooling, is avoided that again to quick The stimulation of sense cortex follicles regions is damaged, at the same can ensure that under liquid nitrogen it is easy to operate accurately, store it is safe and reliable, compatible in order, And the human ovarian of nontoxic pollution-free organizes cryopreservation device under liquid nitrogen.
Technical proposal that the invention solves the above-mentioned problems is:
A kind of human ovarian organizes cryopreservation device under liquid nitrogen, including for deposit human ovarian tissue body, the pipe One end of body is provided with opening, and opening is furnished with closure, and the closure is detachably connected with the body, closure and body Enclose closed parking space;
The inner top surface of the closure is provided with the fork for taking human ovarian's tissue for fork and takes device.
Further, the fork takes device including some fine needles be arrangeding in parallel, and the fixing end of fine needle is vertically fixed on close On the inner face of capping, free end axially the extending into body along body of fine needle.
Further, the fork takes device including 3 fine needles, and 3 fixing ends of 3 fine needles of first place connection constitute one positive three It is angular, and the center of circle of closure inner face is the center of the equilateral triangle.
Further, the material of the fine needle is argent, and the diameter range of the fine needle is 0.3mm~0.4mm.
Further, the body is hollow cylinder shape, and the outside wall surface at body open is provided with external screw thread, closure Internal face is provided with the female thread being engaged with the external screw thread.
Further, the material of the body and the closure is the polypropylene for being resistant to -196 DEG C of low temperature.
Further, a diameter of 26mm of the body, a diameter of 27mm of closure.
Further, sealing ring is provided with the closure, the sealing ring is silicon gum gasket ring.
Fine needle of the present invention is used to fix direct plunge into Liquid Nitrogen after ovary tissue so that tissue fritter can be straight with liquid nitrogen Contact, and fine needle adopts the higher argent of heat conductivity as material, then cause ovary tissue to lower the temperature in liquid nitrogen faster, Glass freezing effect can preferably be reached;The medullary substance part of ovary tissue is fixed with acupuncture needle, then effectively prevent tweezer under liquid nitrogen Sub-folder takes and the physical stimulation of the cortical section of follicle enrichment is damaged.
And the design of inventive closure formula then avoids other pollutions of iatrogenic material to ovary tissue in liquid nitrogen container;Carefully The arrangement of pin is set to equilateral triangle, and fine needle diameter range is set to 0.3mm~0.4mm, then can effectively by ovary tissue A plane is fixed on, and the diameter of body and closure is respectively set to the 2 of conventional 2ml spermatozoa cryopreservation pipe pipe shafts and lid Times, the parking space of ovary tissue had both been ensure that, cause the cryopreservation device to be put into conventional freezing storing box with compatibility again and (cut with blade Go to be connected " cross " plastic support in 4 little square storage element lattice central authorities, that is, make the big pros that capacity is 4 times Shape storage element lattice).
The hickey part colours of closure and body are set to assertive colours (redness) so that in the feelings for having liquid nitrogen to volatilize Both identifications that also can be apparent under condition, and accurately carry out screwing operations;Closure and body are adopted and are resistant to -196 DEG C The polypropylene material of low temperature, and through ethylene oxide sterilization processes, it is to avoid body breach and material caused by ultralow temperature Toxic action to ovary tissue itself.The present invention is repeatable after sterilized to be utilized, and can be made marks on outer wall of the present invention, It is easy to long-term preservation.
Beneficial effects of the present invention are mainly manifested in:
1st, fork of the invention takes device and had both avoided under liquid nitrogen with the gripping of tweezers blindness to follicle enrichment in ovary tissue The physical damnification of cortical section, turn avoid the cross-contamination with other iatrogenic materials in liquid nitrogen container, and fork takes device for solid Determine the medullary substance part of ovary tissue, simple structure, facilitate handy.
2nd, fine needle adopts the higher argent of heat conductivity as material so that ovary tissue can reach faster under liquid nitrogen To glass freezing effect.
3rd, device is repeatable after sterilized utilizes, economical environment-protective.
Description of the drawings
Fig. 1 is the structural representation of the present invention;
Fig. 2 is the schematic diagram after the freezing storing box that the embedded plastic cross shape of pruning of the present invention is supported.
Specific embodiment
Embodiment 1
With reference to Fig. 1, a kind of human ovarian organizes cryopreservation device under liquid nitrogen, including for depositing the body of human ovarian's tissue 3, one end of the body 3 is provided with opening, and opening is furnished with closure 3, and the closure 1 is detachably connected with the body 3, Closure 1 encloses closed parking space with body 3;
The inner top surface of the closure 1 is provided with the fork for taking human ovarian's tissue for fork and takes device.
Embodiment 2
A kind of human ovarian described in embodiment is organized under liquid nitrogen in cryopreservation device:It is flat including some that the fork takes device The fine needle that row is arranged, the fixing end of fine needle is vertically fixed on the inner face of closure, and the free end of fine needle is along the axial direction of body Extend into body.
Embodiment 3
A kind of human ovarian described in embodiment 1 or 2 is organized under liquid nitrogen in cryopreservation device:The fork takes device including 3 Fine needle 2, the length of 3 fine needles 2 is 25mm.3 fixing ends of 3 fine needles 2 of first place connection constitute an equilateral triangle, profit In by ovary tissue fix in one plane;And the center of circle of the inner face of closure 1 is the center of the equilateral triangle so that ovum Nest tissue is fixed on the center position of closure 1.
Embodiment 4
A kind of human ovarian described in embodiment 1-3 any one is organized under liquid nitrogen in cryopreservation device:The material of the fine needle 2 Matter is argent, because silver has higher heat conductivity, is more beneficial for that ovary tissue is very fast to be cooled to glassy state.And it is described The diameter range of fine needle 2 is 0.3mm~0.4mm.
Embodiment 5
A kind of human ovarian described in embodiment 1-4 any one is organized under liquid nitrogen in cryopreservation device:The body 3 is sky Heart cylindrical shape, the outside wall surface at the opening of body 3 is provided with external screw thread 4, and the internal face of closure 1 is provided with and the external screw thread 4 female threads 7 being engaged.Body 3 and closure 1 are realized being detachably connected by female thread 7 and external screw thread 4.Embodiment 6
A kind of human ovarian described in embodiment 1-5 any one is organized under liquid nitrogen in cryopreservation device:The body 3 and institute The material for stating closure 1 is the polypropylene for being resistant to -196 DEG C of low temperature, and through ethylene oxide sterilization processes, to ensure to freeze Cryopreservation device does not ftracture under liquid nitrogen and avoids material itself from causing toxic damages to ovary tissue.
Embodiment 7
A kind of human ovarian described in embodiment 1-6 any one is organized under liquid nitrogen in cryopreservation device:The body it is straight Footpath is 26mm, is 2 times of conventional 2ml sperm freezing pipe pipe shaft diameters;A diameter of 27mm of closure, is that conventional 2ml sperms are cold 2 times of frozen pipe lid diameter, both ensure that the parking space of ovary tissue, cause this invention to be put into conventional jelly with compatibility again Deposit box.Closure sidewall thickness is 0.5mm.
Embodiment 8
A kind of human ovarian described in embodiment 1-7 any one is organized under liquid nitrogen in cryopreservation device:In the closure 1 Sealing ring 5 is provided with, the sealing ring 5 is silicon gum gasket ring, to guarantee the sealing of closure 1 and body 2.
Embodiment 9
A kind of human ovarian described in embodiment 1-8 any one is organized under liquid nitrogen in cryopreservation device:The closure 1 The color of the external screw thread 4 of female thread 7 and body 3 is set to assertive colours (as red), is easy to be identified and dock under liquid nitrogen.
Embodiment 10
A kind of human ovarian described in embodiment 1-9 any one is organized under liquid nitrogen in cryopreservation device:Outside the body 3 Wall is provided with blank marks frame 6, conveniently fills in ovary tissue freezing information.
The present invention use principle be:
Human ovarian's tissue is broadly divided into cortical section and medullary substance part, and the follicle with reproductive function is enriched in entirely Cortical section.When glass freezing is carried out, generally first ovary tissue is cut into into 5-10mm length, 1-2mm thick square tiles, After a series of permeability that concentration gradually rise and impermeability cryoprotective agent immersion treatment, closure 1 is held, will 3 fine needles 2 are pierced into the medullary substance part of ovary tissue, then in liquid nitrogen of the rapid input without other iatrogenic samples of contact, The external screw thread 4 of assertive colours (as red) is identified with by the identification under liquid nitrogen, with the tweezers after prior pre-cooling sealing is clamped respectively The two ends of lid 1 and body 3, will both screwing hermetics.What refrigerating plant was subsequently embedded in a plastic cross shape of pruning supports is normal With (as shown in Figure 2) in freeze box 8, then freezing storing box 8 is put in the grape in liquid nitrogen container, that is, completes whole refrigerating process.
Content described in this specification embodiment is only enumerating to the way of realization of inventive concept, the protection of the present invention Scope is not construed as being only limitted to the concrete form that embodiment is stated, protection scope of the present invention also includes art technology Personnel according to present inventive concept it is conceivable that equivalent technologies mean.

Claims (8)

1. a kind of human ovarian organizes cryopreservation device under liquid nitrogen, it is characterised in that:Including for deposit human ovarian tissue pipe Body, one end of the body is provided with opening, and opening is furnished with closure, and the closure is detachably connected with the body, close Capping encloses closed parking space with body;
The inner top surface of the closure is provided with the fork for taking human ovarian's tissue for fork and takes device.
2. a kind of human ovarian as claimed in claim 1 organizes cryopreservation device under liquid nitrogen, it is characterised in that:The fork takes device Including some fine needles be arrangeding in parallel, the fixing end of fine needle is vertically fixed on the inner face of closure, the free end of fine needle Axially extending into body along body.
3. a kind of human ovarian as claimed in claim 2 organizes cryopreservation device under liquid nitrogen, it is characterised in that:The fork takes device Including 3 fine needles, 3 fixing ends of 3 fine needles of first place connection constitute an equilateral triangle, and the center of circle of closure inner face is The center of the equilateral triangle.
4. a kind of human ovarian as claimed in claim 3 organizes cryopreservation device under liquid nitrogen, it is characterised in that:The material of the fine needle Matter is argent, and the diameter range of the fine needle is 0.3mm~0.4mm.
5. a kind of human ovarian as claimed in claim 4 organizes cryopreservation device under liquid nitrogen, it is characterised in that:The body is sky Heart cylindrical shape, the outside wall surface at body open is provided with external screw thread, and the internal face of closure is provided with and the external screw thread phase The female thread of cooperation.
6. a kind of human ovarian as claimed in claim 5 organizes cryopreservation device under liquid nitrogen, it is characterised in that:The body and institute The material for stating closure is the polypropylene for being resistant to -196 DEG C of low temperature.
7. a kind of human ovarian as claimed in claim 6 organizes cryopreservation device under liquid nitrogen, it is characterised in that:The body it is straight Footpath is 26mm, a diameter of 27mm of closure.
8. a kind of human ovarian as claimed in claim 7 organizes cryopreservation device under liquid nitrogen, it is characterised in that:In the closure Sealing ring is provided with, the sealing ring is silicon gum gasket ring.
CN201611103273.4A 2016-12-05 2016-12-05 Device is deposited to freezing under human ovary tissue liquid nitrogen Active CN106577632B (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108639514A (en) * 2018-06-14 2018-10-12 浙江大学 A kind of cell freezing storage device and its method towards biological 3D printing
CN112741079A (en) * 2020-12-30 2021-05-04 浙江大学 Ovarian tissue sample medulla removing device
WO2021129312A1 (en) * 2019-12-24 2021-07-01 上海明悦医疗科技有限公司 Carrier, vacuumizing device and tissue cryopreservation system

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN2758734Y (en) * 2004-12-16 2006-02-15 昆明医学院 Container suitable for liquid nitrogen cyliner to store frozen cell or tissue
CN101314757A (en) * 2008-07-15 2008-12-03 四川大学华西第二医院 Aculeus type tissue slice glassivation refrigeration carrier and refrigeration method for ovary tissue
CN202522460U (en) * 2012-04-24 2012-11-07 浙江省台州医院 Tissue sample cryopreservation tube
CN203646394U (en) * 2013-10-14 2014-06-18 四川大学华西第二医院 Vitrification carrier

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN2758734Y (en) * 2004-12-16 2006-02-15 昆明医学院 Container suitable for liquid nitrogen cyliner to store frozen cell or tissue
CN101314757A (en) * 2008-07-15 2008-12-03 四川大学华西第二医院 Aculeus type tissue slice glassivation refrigeration carrier and refrigeration method for ovary tissue
CN202522460U (en) * 2012-04-24 2012-11-07 浙江省台州医院 Tissue sample cryopreservation tube
CN203646394U (en) * 2013-10-14 2014-06-18 四川大学华西第二医院 Vitrification carrier

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108639514A (en) * 2018-06-14 2018-10-12 浙江大学 A kind of cell freezing storage device and its method towards biological 3D printing
CN108639514B (en) * 2018-06-14 2023-05-16 浙江大学 Cell freezing storage device and method for biological 3D printing
WO2021129312A1 (en) * 2019-12-24 2021-07-01 上海明悦医疗科技有限公司 Carrier, vacuumizing device and tissue cryopreservation system
CN112741079A (en) * 2020-12-30 2021-05-04 浙江大学 Ovarian tissue sample medulla removing device
CN112741079B (en) * 2020-12-30 2022-07-01 浙江大学 Ovarian tissue sample medulla removing device

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