CN205143336U - Freezing carrier of preserving of trace sperm - Google Patents
Freezing carrier of preserving of trace sperm Download PDFInfo
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- CN205143336U CN205143336U CN201520988161.6U CN201520988161U CN205143336U CN 205143336 U CN205143336 U CN 205143336U CN 201520988161 U CN201520988161 U CN 201520988161U CN 205143336 U CN205143336 U CN 205143336U
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- protective carrier
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Abstract
The utility model discloses a freezing carrier of preserving of trace sperm, freezing save carrier are rectangle, square or circular shape level form or recess form, the refrigerating fluid droplet of size for bearing a 1 -3 0.1 -10.0uL to transparent plastic materials or transparent glass material producing. Freezing preserve insert the sperm culture dish of lid oil on the carrier behind the ready refrigerating fluid droplet in, snatch the sperm with the microinjection needle and put into the refrigerating fluid droplet, take out the freezing carrier of preserving and use simultaneously and absorb water paper and blot mineral oil, quick freezing, will freeze save carrier disect insertion ICSI ware when unfreezeing, utilize its 37 DEG C of mineral oil recovery sperms. The utility model discloses to freeze and preserve that to grab the sperm in the sperm culture dish that the carrier inserted lid oil freezing, need not the special device of moisturizing, not worry the evaporation of refrigerating fluid droplet, operate extremely simple, very big improvement azoospermia patient assist the flexibility and the reliability of fertility treatment, the sperm rate of recovery can reach 100%, sperm survival rate about 60%.
Description
Technical field
The utility model relates to the assisted reproduction field in medical science, more particularly, relates to the freezen protective carrier of micro-sperm (containing single sperm, the sperm etc. as testicular biopsy sperm, epididymis puncture sperm, severe oligospermia patient).
Background technology
Supplementary reproduction clinically, azoospermia patients generally will carry out the diagnostic puncture of epididymis or testis, to guarantee that the ovulation induction cycle can obtain enough sperms and treat for intracytoplasmic sperm injection (Intracytoplasmicsperminjection, ICSI).Due to the pregnancy rate only about 50% of each ICSI treatment cycle, in order to obtain gestation, some patients needs to repeat ICSI treatment for several times.In order to reduce as much as possible patient repeatedly puncture get essence misery, protect holandric spermatogenesis better, medical worker attempt to azoospermia patients diagnostic puncture obtain or the remaining micro-motile sperm of ICSI treatment cycle carry out freezen protective, later the wife's side get ovum day by recovery after the sperm of surviving be used for ICSI, this will improve flexibility and reliability that such patient's assisted reproduction treats greatly.The sperm of severe oligozoospermia patient is unstable, and the sperm of preventative such patient freezing is also very necessary, this will avoid the wife's side get ovum day without sperm can with and cancel the cycle.
Trace sperm cryopreservation is the difficult problem in supplementary reproduction field.Traditional sperm freezing adopts the cryovial (Cryovial) of 1.8mL as freezing carrier, and freezing/resuscitation process needs to make 2-4 centrifuge washing to sperm, and every single stepping all can cause sperm loss in various degree.And the general quantity of sperm that testis or epididymis puncture obtain is few, vigor is poor, after conventional cryopreservation recovery, be often difficult to find enough motile sperms to carry out ICSI, so the sperm that traditional sperm freezing method is not suitable for puncture obtains carries out freezen protective.
Chinese patent application 200510061871.5 provides the frozen of a kind of testicular sperm and method for resuscitation; the oolemma that this invention is found time with endochylema is for carrier; the testicular sperm of biopsy is loaded in the oolemma closed; carry out interpolation and the wash-out of sperm freezing protecting agent; in cryopreservation resuscitation process, sperm Seal and preservation is in oolemma; in the method, the recovery of oolemma sperm need not be separated and washing centrifugal process through cumbersome discontinuous density gradient; can direct sucking-off sperm under micromanipulation instrument, be directly used in ICSI.But the method needs the oolemma of animal (mouse or rat) ovum or people's ovum, material source difficulty (human oocyte zona pellucida is difficult to obtain), and there is dispute of ethic (during ICSI, host DNA has the risk being injected into people's ovum together with sperm), be difficult to be widely used in supplementary reproduction in this way clinical.
A kind of freezing and defreezing method of micro-sperm and the freezing and thawing apparatus (Chinese patent application 200810042695.4) of sperm were once invented by this seminar, freezing of the lengthy motion picture shape that this invention is made with glass or polystyrene or channel-shaped is loaded sperm, the droplet that sperm is placed on freezing is freezing, thaw in deep fat, the sperm rate of recovery is close to 100%, but the method needs the dedicated wet box manufacturing saturated humidity condition, to ensure that in refrigerating process, droplet does not evaporate, loaded down with trivial details, the consuming time length of operating process, this strongly limits this invention widely using clinically.
The object of this invention is to provide simple to operate, micro-sperm is (containing single sperm efficiently, sperm etc. as testicular biopsy sperm, epididymis puncture sperm, severe oligospermia patient) freezen protective carrier, in order to solve this difficult problem of micro-sperm freezing, Yangjing 2008.09.09 rectify, Peng Qiu equality application patent of invention: the freezing and defreezing method of sperm and the freezing and thawing apparatus of sperm, this license notification number CN101671651B, the object of this invention loads the micro-sperm such as testicular sperm, epididymal sperm with freezing to carry out freezing.During freezing freezing micro-sperm, need by micromanipulation system, single sperm one rule to be transferred on freezing, freezing liquid droplet volume used very little (0.5-3.0 μ L), and the time expended long (10-30 minute).Evaporate to prevent droplet and change the osmotic pressure of freezing liquid, so the process need of whole crawl sperm completes under the condition of constant humidity, namely freezing is placed in wet box and operates.
In order to provide stable, reliable constant humidity condition, 2011.06.07 Lu Hui, Peng Qiu equality application utility model patent: a kind of moisturizing device (Authorization Notice No. CN202122042U of rare sperm freezing, authorized announcement date 2012.01.25), this moisturizing device only there is provided the condition of constant humidity, the humidity shifting environment can be ensured in the transfer process of sperm freezing, prevent in process sperm being transferred to from culture dish freezing carrier (as freezing) because drop evaporation makes the osmotic pressure outside sperm increase, cause dead sperm, reduce anabiosis rate.This moisturizing device itself is not the freezing carrier loading sperm.
Summary of the invention
First object of the present utility model is to provide a kind of simple and quick micro-sperm (containing single sperm, the sperm etc. as testicular biopsy sperm, epididymis puncture sperm, severe oligospermia patient) freezen protective carrier.
For realizing above first object, the utility model discloses following technical scheme: a kind of micro-sperm cryopreservation carrier, it is characterized in that, described freezen protective carrier is rectangle, square or circular horizontal or groove-like, size is the freezing liquid droplet that can carry 1-3 0.1-10.0uL, make with transparent plastic material or transparent glass material, at least one end margin of described freezen protective carrier arranges protruding bare terminal end, when freezen protective carrier is rectangle, size is long 0.3-1.5cm, wide 0.2-1.0cm, when freezen protective carrier is rounded, size is diameter 0.2-1.0cm.
The utility model has the advantage of: (1) freezing liquid is same with conventional sperm freezing liquid phase, has no special requirements, can conveniently obtain, as commercially available glycerinated sperm freezing liquid.(2) grab sperm freezing by the sperm culture dish of freezen protective carrier insertion lid oil, without the need to special moisturizing device, do not worry that freezing liquid droplet evaporates, the sperm having time enough as far as possible many with patient crawl like this carries out freezing.The motile sperm of the sperm that puncture obtains or severe oligozoospermia patient is very rare, needs to spend the more time could capture abundant motile sperm and carries out freezen protective.(3) thaw in the ICSI ware of freezen protective carrier insertion lid oil, without the need to centrifuge washing, can be directly used in ovum microscopic insemination, sperm can not be lost, and the sperm rate of recovery can reach 100%.(4) before freezing, the mineral oil blotting paper on freezen protective carrier can be blotted as far as possible, this ensure that the anabiosis rate of sperm is not by the negative effect of mineral oil, after recovery, the survival rate of sperm can reach more than 60%.(5) whole freeze-thaw operation is very simple, as long as can carry out microscopic insemination just can use the freezing single sperm of the method, is convenient to extensively promote clinically in supplementary reproduction.(6) the utility model solves a difficult problem that is complicated, loaded down with trivial details in current monosperm Cryopreservation Technology, inefficiency well, greatly improves flexibility and reliability that azoospermia patients and severe oligozoospermia patient accept assisted reproduction treatment.
Accompanying drawing explanation
Fig. 1 is rectangle freezen protective carrier schematic diagram.
Freezen protective carrier positions schematic diagram when Fig. 2 is freezing.
Fig. 3 is the position view of freezen protective carrier in ICSI ware when thawing.
Label in Fig. 1 is:
1---freezen protective carrier; 11---bare terminal end.
Embodiment
Below in conjunction with specific embodiment, set forth the utility model further.The experimental technique used in following embodiment if no special instructions, is conventional method.Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.Should be understood that these embodiments are only not used in restriction scope of the present utility model for illustration of the utility model.
The micro-sperm cryopreservation carrier of embodiment 1.
Refer to Fig. 1, Fig. 1 is a preferred embodiment of the present utility model, freezen protective carrier (also known as freezing) be rectangle, size is the freezing liquid droplet that can carry 1-3 0.1-10.0uL, make with transparent plastic material (as polystyrene or polypropylene) or transparent glass material, an end margin arranges protruding bare terminal end.
As a preferred version, described freezen protective carrier is rectangle, and size is long 0.3-1.5cm, wide 0.2-1.0cm.
The shape of freezen protective carrier can be rectangle, square or circular horizontal or groove-like, as long as be convenient to be kept at liquid nitrogen container for a long time, follow-up refrigerating process is carried out, as freezen protective carrier inserted the cryovial of 0.5-2.0mL or being connected in rod-shaped material by freezen protective carrier in the container that freezen protective carrier can be stored in arbitrary shape or on backing material.As long as size can carry the freezing liquid droplet of 1-3 0.1-10.0uL, the variable dimension of freezen protective carrier, for freezen protective carrier storage in the cryovial of 0.5-2.0mL, its size can be the arbitrary dimension of the cryovial (Vial) can easily putting into 0.5-2.0mL.
Conveniently freezen protective carrier is operated, as put into/taking out the storage container (as 0.5-2.0mL cryovial) etc. of sperm culture dish, ICSI ware and freezen protective carrier, freezen protective carrier is designed with the little tooth L-shaped with freezen protective carrier sheet part or the projection of similar holding thing, can clamps for pincet.
Freezing and the defreezing method of the micro-sperm of embodiment 2.
Time freezing, first prepare sperm culture dish, namely make long drop with 10%SSS-mHTF and cover mineral oil.Inserted in a small amount of 10%SSS-mHTF by the convoluted seminiferous tubule that biopsy is taken out, convoluted seminiferous tubule pulverizes through elbow tweezers, directly adds the long 10%SSS-mHTF drop in sperm culture dish.Freezen protective carrier prepares the freezing liquid droplet of 15 microlitre, freezen protective carrier is inserted in the sperm culture dish of lid oil, the freezing liquid droplet on freezen protective carrier is inserted with the motile sperm that injection needle (ICSIInjectionPipette) captures some (5-10 bar sperm/sheet), freezen protective carrier is taken out with pincet, and blot the mineral oil on freezen protective carrier with blotting paper after, put into the cryovial (Vial) of 1.8mL as far as possible, snap frozen is carried out by the order of the liquid nitrogen vapor 5 minutes → input liquid nitrogen of 5cm height above 4 DEG C of 20 minutes →-20 DEG C 20 minutes → liquid nitrogen surfaces, long-term preservation.
During recovery, ovum day is got on the wife's side, i.e. ICSI same day, cryovial (Vial) is taken out in liquid nitrogen container, taken out in cryovial by freezen protective carrier with pincet, and insert in ICSI ware (containing 37 DEG C of mineral oil) rapidly, under inverted microscope, (100-200 times of multiplication factor) rakes about sperm in freezing liquid droplet, and proceed in mHTF droplet with injection needle, for follow-up immobilization of spermatozoa and mature egg ICSI.
Freezen protective carrier positions schematic diagram when Fig. 2 is freezing, the long drop of several 10%SSS-mHTF is prepared in ware the first half, cover mineral oil, the freezing puncture sperm of preparation is added this long drop, freezen protective carrier is placed on below long drop, i.e. ware the latter half, inserts sperm crawl in the freezing liquid droplet on freezen protective carrier with micromanipulation entry needle.
Fig. 3 is the position view of freezen protective carrier in ICSI ware when thawing, the drop at nine grids center is 10%PVP, braking sperm is used, other 8 drops are 10%SSS-mHTF, treat that the ovum of ICSI is inserted wherein, the little drop in the right is also 10%SSS-mHTF, is used for the sperm that rinse finds from freezen protective carrier, sperm is in this droplet after rinse, and the 10%PVP dripping being transferred to center is employed in ICSI.
This freezing carrier just in time can be placed in the conventional sperm freezing pipe (Vial) of commercially available 1.0-1.5mL, very convenient in storing in liquid nitrogen container.And, at least one end margin of described freezen protective carrier arranges protruding bare terminal end, can insert very easily in the oily sperm culture dish of lid and directly grab sperm freezing, without the need to special moisturizing device, do not worry freezing liquid droplet evaporation (having mineral oil to cover), have time enough and the patient many sperms of trying one's best that capture to carry out freezing.In addition, the constant humidity condition not needing dependence moisturizing device to provide, also makes the operation of crawl sperm become simpler, convenient, is easy to promote micro-sperm freezing technology clinically.
The above is only preferred embodiment of the present utility model; should be understood that; for those skilled in the art; under the prerequisite not departing from the utility model principle; some improvements and modifications can also be made; such as the shape of freezing, size are suitably beautified and adjusted, or suitably revising bare terminal end, protection domain of the present utility model must be considered as.
Claims (1)
1. a micro-sperm cryopreservation carrier, it is characterized in that, described freezen protective carrier is rectangle, square or circular horizontal or groove-like, size is the freezing liquid droplet that can carry 1-3 0.1-10.0uL, make with transparent plastic material or transparent glass material, at least one end margin of described freezen protective carrier arranges protruding bare terminal end, when freezen protective carrier is rectangle, size is long 0.3-1.5cm, wide 0.2-1.0cm, when freezen protective carrier is rounded, size is diameter 0.2-1.0cm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201520988161.6U CN205143336U (en) | 2015-05-19 | 2015-12-03 | Freezing carrier of preserving of trace sperm |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2015203227303 | 2015-05-19 | ||
| CN201520322730 | 2015-05-19 | ||
| CN201520988161.6U CN205143336U (en) | 2015-05-19 | 2015-12-03 | Freezing carrier of preserving of trace sperm |
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| CN205143336U true CN205143336U (en) | 2016-04-13 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201520988161.6U Expired - Fee Related CN205143336U (en) | 2015-05-19 | 2015-12-03 | Freezing carrier of preserving of trace sperm |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110521720A (en) * | 2019-01-18 | 2019-12-03 | 上海廪典实业有限公司 | A kind of superfreeze and closed store method |
| WO2021018292A1 (en) * | 2019-08-01 | 2021-02-04 | 常州市妇幼保健院 | Method for storing individual sperm or trace amount of sperm, and chip carrier |
-
2015
- 2015-12-03 CN CN201520988161.6U patent/CN205143336U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110521720A (en) * | 2019-01-18 | 2019-12-03 | 上海廪典实业有限公司 | A kind of superfreeze and closed store method |
| WO2021018292A1 (en) * | 2019-08-01 | 2021-02-04 | 常州市妇幼保健院 | Method for storing individual sperm or trace amount of sperm, and chip carrier |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160413 Termination date: 20161203 |