CN108935445A - A kind of jelly precision method based on ICSI micromanipulation - Google Patents
A kind of jelly precision method based on ICSI micromanipulation Download PDFInfo
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- CN108935445A CN108935445A CN201810897724.9A CN201810897724A CN108935445A CN 108935445 A CN108935445 A CN 108935445A CN 201810897724 A CN201810897724 A CN 201810897724A CN 108935445 A CN108935445 A CN 108935445A
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- sperm
- icsi
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION 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
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
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Abstract
The invention belongs to medicine technology fields, are related to a kind of jelly precision method based on ICSI micromanipulation.Method includes the following steps: (1) prepares two ICSI wares, a placement sperm sample, another is placed 1 to few drops sperm freezing and protects liquid；(2) under ICSI inverted microscope, motile sperm is drawn from the ICSI ware for placing sperm sample with intracytoplasmic sperm injection needle, is placed in the drop of sperm freezing protection liquid of another ICSI ware；(3) the ICSI ware of the drop containing sperm freezing protection liquid is placed in liquid nitrogen and freezes and saves.Method of the invention is not necessarily to carrier, significantly reduces intermediate link of a small amount of sperm during freezing and resuscitation, and sperm motility height, sperm Loss Rate are low after defrosting, and the utilization rate of sperm is high.
Description
Technical field
The invention belongs to medicine technology fields, more particularly, to a kind of jelly precision method based on ICSI micromanipulation.
Background technique
Frozen sperm is required technology in supplementary reproduction, and in vitro in fertilization process, some patientss are because of motile sperm
Number is less or takes the reasons such as smart difficulty, needs frozen sperm in advance, in case use in vitro fertilization.Part of patient activity's sperm
Number it is less, conventional method freezing operation link it is more, in operating process there are many utensil and prepare freezing or thaw
Sperm and its cryoprotector are in contact, and are easy to cause a small amount of sperm loss for preparing freezing or defrosting, are difficult to find after defrosting
Motile sperm.In clinical position, the number of motile sperm person below 10-15 item, is freezed with conventional method before freezing, and is thawed
After be difficult to find motile sperm.
The freezing of the existing a small amount of sperm of the mankind is using the smart technology of micro- jelly.Detailed process is will to contain a small amount of sperm
And its cryoprotector is packed into straw, then sets in liquid nitrogen the straw containing a small amount of sperm and its cryoprotector and freezes；Such as
Thaw sperm, then is removed from liquid nitrogen straw, after rewarming by straw a small amount of sperm and its cryoprotector be sucked out, set
It is spare in ware (ICSI ware).
Intracytoplasmic sperm injection (intracytoplasmic sperm injection, ICSI) is widely used in
Treat male sterility；ICSI micromanipulation system is used for the stored frozen of sperm, is mainly drawn from ICSI ware with injection needle
Motile sperm, and the sperm of absorption is placed on the carriers such as oolemma, cryopreservation.The stored frozen mode of sperm has suction pipe
Method, ampulla method, ice crystal method and oolemma store 4 kinds of method.In oolemma storage method, according to ICSI operating process, sperm is infused
Enter oolemma, an oolemma storage is no more than 5 sperms, the oolemma with sperm is placed in straw or straw, freezing is protected
It deposits, the oolemma containing sperm is put into spare in ICSI ware by when defrosting.Though there is ICSI to draw sperm both at home and abroad to be freezed,
Its carrier freezed is straw, capsule or oolemma.CN108207931A discloses a kind of micro sperm cryopreservation method,
Though straw, capsule or oolemma are not used in, but still uses hydrogel microsphere as carrier, when defrosting need to be by sperm and its protection
Agent is sucked out from carrier, and drop, which is made, to be used, and increases too many micromanipulation and intermediate link.
Therefore, existing a small amount of smart technical operation link of sperm jelly is more, and operating process is there are many utensil and prepares to freeze
Or the sperm and its cryoprotector to thaw is in contact, and is easy to cause a small amount of sperm for preparing to freeze or thaw to lose, and thaw
Needing for sperm and its cryoprotector to be transferred to certain vessel rear afterwards can be used, and increase intermediate ring in operation
Section, hence it is evident that increase a possibility that sperm is lost and inactivated.Meanwhile the expense and increase of purchase carrier are also added using carrier
The technical difficulty of operator and operating time
Summary of the invention
The object of the present invention is to provide a kind of jelly precision methods based on ICSI micromanipulation.Method of the invention is without carrying
Body significantly reduces intermediate link of a small amount of sperm during freezing and resuscitation, sperm motility height, sperm after defrosting
Loss Rate is low, and the utilization rate of sperm is high.
Specifically, the present invention provides a kind of jelly precision method based on ICSI micromanipulation, method includes the following steps:
(1) prepare two ICSI wares, one is placed sperm sample (puncture fluid including projecting sperm and testis and epididymis),
Another is placed 1 to few drops sperm freezing and protects liquid；
(2) under ICSI inverted microscope, with the absorption activity from the ICSI ware for placing sperm sample of intracytoplasmic sperm injection needle
Sperm is placed in the drop of sperm freezing protection liquid of another ICSI ware；
(3) the ICSI ware of the drop containing sperm freezing protection liquid is placed in liquid nitrogen and freezes and saves.
The present invention draws sperm and is directly placed into ICSI ware and freezes using ICSI puncture needle, to further take out observation or application
It cuts down the number of intermediate links.
In the present invention, the sperm freezing protection liquid can be conventional various self-controls or the sperm freezing protection of commercialization
Liquid, the present invention are not particularly limited this.A kind of specific embodiment according to the present invention, the sperm freezing protection liquid is commodity
The sperm freezing of change protects liquid (Quinns AdvantageTMSperm Freezing Medium)。
The present invention be a kind of micromanipulation system by ICSI based on micro- jelly essence technology, be a kind of improved micro- jelly
Smart technology.It is composed of suction essence technology two parts in conventional method frozen sperm technology and Intracytoplasmic sperm injection.Respectively
Routine operation process can be used in part.
According to the present invention, step (3) can also include: the ICSI ware by the drop containing sperm freezing protection liquid in 2-6
DEG C place 10-20 minutes, be followed by placed in liquid nitrogen vapor 20-30 minutes, be finally placed in liquid nitrogen and freeze.The step directly freezed
Drop containing sperm freezing protection liquid is not needed to be sucked out on the drop to carrier of sperm and its cryoprotector, then be freezed
Carrier and its drop reduce the link that sperm to carrier is shifted when freezing.
According to the present invention, when defrosting sperm, the ICSI ware of the drop containing sperm freezing protection liquid is removed from liquid nitrogen,
It is spare after rewarming.The specific steps of the rewarming include: the ICSI ware by the drop containing sperm freezing protection liquid from liquid nitrogen
It takes out, is quickly put into 37 DEG C of water baths recovery 5-10 minutes, it is directly spare.It does not need that sperm and its freezing is sucked out in the step
Protective agent is spare to ICSI ware, but directly can be spare, reduces the link that sperm is shifted after thawing.
Using method of the invention, new instrument, equipment and reagent not will increase, instrument used, equipment and reagent are not
It can poison and the sperm of injured patient, any risk will not be brought to patient.Compared with conventional method, jelly essence side of the invention
Method, sperm motility is higher after defrosting, and sperm Loss Rate is lower；It, can with jelly precision method of the invention to the patient for having indication
It is obviously improved its Clinical Outcome.
Method of the invention can be used for a small amount of sperm of the existing mankind (including ejaculated sperm, testis and epididymal sperm etc.) and essence
The freezing for staying in the remaining sperm in ICSI ware in sub- endochylema after microinjection, the number especially suitable for motile sperm before freezing
The person below 10-15 item.In addition, method of the invention applies also for the freezing of the sperm of row testicular biopsy art acquirement, pass through this
The sperm of acquirement is divided into more parts of freezings, takes portion to be fertilized every time, can significantly reduce patient because of failure in vitro fertilization by technology
Row testicular biopsy art takes smart probability again afterwards, reduces the pain and financial burden of patient.
Other features and advantages of the present invention will then part of the detailed description can be specified.
Specific embodiment
The preferred embodiment of the present invention is described in more detail below.Although the following describe preferred implementations of the invention
Mode, however, it is to be appreciated that may be realized in various forms the present invention without that should be limited by the embodiments set forth herein.
Embodiment
It is tested using Guizhou affiliated hospital of medical university 6 parts of discarded semen samples of reproductive center.Including following step
It is rapid:
(1) prepare two ICSI wares, a placement sperm sample, another is placed 1 to few drops sperm freezing and protects liquid；
(2) under ICSI inverted microscope, with the absorption activity from the ICSI ware for placing sperm sample of intracytoplasmic sperm injection needle
Sperm is placed in the drop of sperm freezing protection liquid of another ICSI ware；
(3) the ICSI ware of the drop containing sperm freezing protection liquid is placed 15 minutes in 4 DEG C, is followed by placed on liquid nitrogen vapor
In 25 minutes, be finally placed in liquid nitrogen and freeze and save；
The sperm freezing protection liquid is Quinns AdvantageTMSperm Freezing Medium。
When defrosting sperm, the ICSI ware of the drop containing sperm freezing protection liquid is removed from liquid nitrogen, it is spare after rewarming,
The step of rewarming includes: to be removed from liquid nitrogen the ICSI ware of the drop containing sperm freezing protection liquid, is quickly put into 37
Recovery 10 minutes in DEG C water bath.
Freeze the success or not of smart technology, is mainly calculated according to sperm motility after jelly essence defrosting and sperm Loss Rate.Freeze
Sperm motility is higher after essence is thawed, and sperm Loss Rate is lower, freezes smart technology and just succeeds.It is as shown in table 1 to freeze smart effect.
1 the method for the present invention frozen sperm effect of table
Note: the sperm number of sperm motility=movable sperm number/total；Sperm before sperm Loss Rate=freezing
The sperm number before sperm number/freezing after number-defrosting；Since what is freezed when conventional method frozen sperm is work in sample
Dynamic sperm and inactive sperm, in order to preferably compared with reproductive center conventional method frozen sperm effect, this method this
What experiment freezed is also the motile sperm and inactive sperm in sample.
Effect analysis after 6 parts of sample freezings and defrosting: sperm mean activity rate is 59.58% before freezing；It is smart after defrosting
The average Loss Rate of son is 3.32%, and sperm mean activity rate is 49.08%.Guizhou affiliated hospital of medical university reproductive center is with often
Rule method frozen sperm, decline 16% before the mean activity rate of sperm relatively freezes after defrosting.With jelly precision method of the invention, thaw
The mean activity rate of sperm only declines 10.4% before relatively freezing afterwards.As it can be seen that compared with routinely freezing precision method, with method of the invention
Freeze essence, sperm motility is higher after defrosting, and sperm Loss Rate is lower.
Various embodiments of the present invention are described above, above description is exemplary, and non-exclusive, and
It is not limited to disclosed each embodiment.Without departing from the scope and spirit of illustrated each embodiment, for this skill
Many modifications and changes are obvious for the those of ordinary skill in art field.
Claims (5)
1. a kind of jelly precision method based on ICSI micromanipulation, which is characterized in that method includes the following steps:
(1) prepare two ICSI wares, a placement sperm sample, another is placed 1 to few drops sperm freezing and protects liquid；
(2) under ICSI inverted microscope, motile sperm is drawn from the ICSI ware for placing sperm sample with intracytoplasmic sperm injection needle,
It is placed in the drop of sperm freezing protection liquid of another ICSI ware；
(3) the ICSI ware of the drop containing sperm freezing protection liquid is placed in liquid nitrogen and freezes and saves.
2. the jelly precision method according to claim 1 based on ICSI micromanipulation, wherein sperm freezing protection liquid is
Quinns AdvantageTMSperm freezing liquid.
3. the jelly precision method according to claim 1 based on ICSI micromanipulation, wherein step (3) further include: will contain
Sperm freezing protect liquid drop ICSI ware in 2-6 DEG C placement 10-20 minutes, be followed by placed in liquid nitrogen vapor 20-30 minutes,
It is finally placed in liquid nitrogen and freezes.
4. the jelly precision method according to claim 1 based on ICSI micromanipulation, wherein when defrosting sperm, essence will be contained
The ICSI ware of the drop of sub- frozen solution is removed from liquid nitrogen, spare after rewarming.
5. the jelly precision method according to claim 4 based on ICSI micromanipulation, wherein the step of rewarming includes:
The ICSI ware of drop containing sperm freezing protection liquid is removed from liquid nitrogen, is quickly put into 37 DEG C of water baths recovery 5-10 points
Clock.
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| CN201810897724.9A CN108935445A (en) | 2018-08-08 | 2018-08-08 | A kind of jelly precision method based on ICSI micromanipulation |
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| CN201810897724.9A CN108935445A (en) | 2018-08-08 | 2018-08-08 | A kind of jelly precision method based on ICSI micromanipulation |
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Citations (6)
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| CN101671651A (en) * | 2008-09-09 | 2010-03-17 | 上海交通大学医学院附属第九人民医院 | Sperm freezing and unfreezing method and sperm freezing and unfreezing device |
| WO2010056755A2 (en) * | 2008-11-11 | 2010-05-20 | Craig H Randall | Microfluidic embryo and gamete culture systems |
| CN104823966A (en) * | 2015-05-19 | 2015-08-12 | 上海交通大学医学院附属第九人民医院 | Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof |
| CN105660607A (en) * | 2016-03-18 | 2016-06-15 | 冯贵雪 | Fertility preserving method of completely immovable sperms |
| CN205884526U (en) * | 2016-08-04 | 2017-01-18 | 江西紫星生物技术有限公司 | Freezing ware of glass ization |
| CN206666552U (en) * | 2017-03-27 | 2017-11-24 | 柳州市妇幼保健院 | A kind of constant temperature egg mother cell/embryo cryopreservation/thawing apparatus |
-
2018
- 2018-08-08 CN CN201810897724.9A patent/CN108935445A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101671651A (en) * | 2008-09-09 | 2010-03-17 | 上海交通大学医学院附属第九人民医院 | Sperm freezing and unfreezing method and sperm freezing and unfreezing device |
| WO2010056755A2 (en) * | 2008-11-11 | 2010-05-20 | Craig H Randall | Microfluidic embryo and gamete culture systems |
| CN104823966A (en) * | 2015-05-19 | 2015-08-12 | 上海交通大学医学院附属第九人民医院 | Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof |
| CN105660607A (en) * | 2016-03-18 | 2016-06-15 | 冯贵雪 | Fertility preserving method of completely immovable sperms |
| CN205884526U (en) * | 2016-08-04 | 2017-01-18 | 江西紫星生物技术有限公司 | Freezing ware of glass ization |
| CN206666552U (en) * | 2017-03-27 | 2017-11-24 | 柳州市妇幼保健院 | A kind of constant temperature egg mother cell/embryo cryopreservation/thawing apparatus |
Non-Patent Citations (3)
| Title |
|---|
| N. BOUAMAMA等: "Comparison of two methods of cryoconservation of sperm when in very small numbers", 《GYNÉCOLOGIE OBSTÉTRIQUE & FERTILITÉ》 * |
| 侯建文等: "单精子冷冻研究进展", 《中华男科学杂志》 * |
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Application publication date: 20181207 |
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