CN102283718A - Preantral follicle separation method and special scalpel thereof - Google Patents

Abstract

The invention discloses a preantral follicle separation method and a special scalpel thereof. The scalpel which is provided in the invention and is used for separating preantral follicles at least comprises two stacked blades, wherein a clearance is left between the blades. The blades are double edge blades. A partition handhold part is respectively arranged at the middle part of each double edge blade along the longitudinal direction; and the partition handhold part is formed by winding adhesive tapes. The scalpel which is used for separating the preantral follicles can be widely applied in separating the preantral follicle outside the body, and has the following advantages of simplicity, high efficiency, low cost and convenience in popularization, after the scalpel is adopted, the number of the separated preantral follicles can be increased, and the separation effect is good.

Description

The method and the special-purpose cutter thereof of follicle before a kind of disengagement chamber

Technical field

The present invention relates to Animal Science (animal reproduction), relate in particular to the method and the special-purpose cutter thereof of the preceding follicle of a kind of disengagement chamber.

Background technology

Tens thousand of primordial follicles are arranged on the cattle ovary, in its growth and maturity process, a considerable amount of follicles degenerations, locking are arranged, the follicular development that only a few is only arranged is to the maturation period, until ovulation.In the female reproduction resource, embryo biotechnologies such as oocyte is that external fertilization, embryo transfer, sex controll, embryo are cut apart, animal cloning and transgenic are researched and developed requisite material, the deficient principal element that becomes these Research progress of restriction in oocyte source, so the exploitation of the preceding follicle of ovarian cavity solves the effective way of this difficult problem beyond doubt.

In recent years, many scholars are devoted to the research of follicle separation method before the chamber, and follicle had been obtained reasonable effect before usefulness enzymes such as Figueiredo separated the cattle chamber in conjunction with Mechanical Method, but it is longer to handle required time with enzyme, and easily follicle before the chamber are caused certain infringement; And Mechanical Method is owing to need special instrument, and involves great expense, and can not excellent popularization come.Therefore, set up apparent very necessary of follicle separation method before the cover simple and direct effective cattle chamber.

Summary of the invention

An object of the present invention is to provide a kind of cutter that is used to separate follicle before the mammal chamber.

The cutter that is used to separate follicle before the mammal chamber provided by the invention comprises at least two blades that stack, and is provided with the gap between the described blade.

Simple in order to use, described blade can be double-edged blade.

In order to produce the gap, longitudinally can be provided with the interval at the middle part of described double-edged blade and hold parts;

For the ease of holding, it is one that the interval of each described double-edged blade holds parts.

For simple to operate, reduce cost, described interval holds that parts can twine form with adhesive tape.

For easy to use, and good separating effect, the width in above-mentioned gap is identical or different.

In order to improve isolating effect, the width in above-mentioned gap is identical, and the width in described gap is 0.056-0.065cm; During actual the use, the width in described gap is specially 0.058cm.

Above-mentioned blade specifically can be 6.

Another object of the present invention provides a kind of method of separating follicle before the mammal chamber of exsomatizing.

Method provided by the invention comprises the steps:

Draw from the cortex portion of the Mammalian Ovary that exsomatizes with above-mentioned cutter and to get, obtain drawing and get thing, promptly obtain follicle before the chamber.

Described stroke of mode of getting got for drawing along horizontal and vertical both direction successively;

Before described stroke is got, also comprise the step of described stripped Mammalian Ovary being removed mesentery, corpus luteum and medullary substance;

Described stroke get after, comprise the steps: also described stroke got that thing once filters successively, secondary filter, the filtrate that obtains is centrifugal, get precipitation, obtains follicle before the chamber.

Filtering the described first time and adopting size is (180-220) μ m mesh screen, filters the described first time and adopts size to be specially 200 μ m mesh screens;

Filtering the described second time and adopting size is 80-120 μ m mesh screen, filters the described second time and adopts size to be specially 100 μ m mesh screens;

Described centrifugal rotation speed is 1450-1550r/min, and described centrifugal rotation speed is specially 1500r/min; The described centrifugal time is 4.5-5.5min, and the described centrifugal time is specially 5min, and described centrifugal radius of turn is 6.02cm;

Before each described filtration, also comprise and blow and beat the step of disperseing to be used for filtering mixture.

Described mammal is a cattle.

Of the present inventionly be used for that the cutter of follicle can be widely used in follicle external isolated ex vivo chamber before before the disengagement chamber, have the following advantages: be simple and direct, efficient, cost is low, be convenient to popularization.Of the present invention experiment showed, by cutter of the present invention adopted separation method provided by the invention, can make isolated chamber before follicle quantity increase and good separating effect.

Description of drawings

Fig. 1 is the structural representation of the cutter of follicle before the disengagement chamber

The specific embodiment

Employed experimental technique is conventional method if no special instructions among the following embodiment.

Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.

The preparation of follicle separating knife before embodiment 1, the cattle chamber

As shown in Figure 1, follicle separating knife 1 before the cattle of the present invention chamber, it comprises that 6 separate the blade 2 that stacks by the gap, are provided with gap 4 between the described blade.Described blade is double-edged blade (a twolip razors slice).Longitudinally be provided with the interval at the middle part of described double-edged blade and hold parts 3; Described interval holds parts and twines formation with adhesive tape; It is one that the interval of each described double-edged blade holds parts.The width in described gap is identical.The width in described gap is 0.058cm.

The preparation method of follicle separating knife is as follows before the cattle of the present invention chamber: get 6 razor blades, twine the middle part of blade one by one with cellophane adhesive tape, make the certain slit (width in slit is 0.058cm) of generation between the blade, to be used to hold with the middle part that cellophane adhesive tape twines all blades again, make as shown in Figure 1.

It is as follows that the present invention is used for before the chamber using method of the isolating cutter of follicle:

1, the obtaining of follicle before the chamber

The cattle ovary that exsomatizes that the slaughterhouse is obtained places the glass dish of 60mm diameter, remove unnecessary mesentery and corpus luteum, subsequently ovary is cut in half along hilum of ovary, remove medullary substance, after in culture medium, cleaning 2 times, use to draw from ovarian cortex portion according to horizontal and vertical both direction and get follicle by the isolating cutter 1 of follicle before the chamber, follicle and the culture medium obtained are blown and beaten with dropper, blow and beat 80～100 times with the suction pipe of diameter 1mm then, filter through 200 μ m mesh screens, reuse diameter 0.5mm suction pipe piping and druming 80-100 time is filtered with 100 μ m mesh screens, with the centrifugal 5min of 1500r/min (centrifugal radius 6.02cm), remove supernatant, precipitation is follicle, and its precipitation adds separating medium dilute suspension, microscopy follicle under the stereomicroscope, counting is also measured diameter;

2, follicle quantity statistics

The above-mentioned follicle that obtains is carried out quantity statistics, determine the efficient that follicle obtains, experiment triplicate, results averaged.

Each ovary is 130 pieces as a result.

Claims (10)

1. a cutter that is used to separate follicle before the mammal chamber comprises at least two blades that stack, and is provided with the gap between the described blade.

2. cutter according to claim 1 is characterized in that: described blade is a double-edged blade.

3. cutter according to claim 1 and 2 is characterized in that:

Longitudinally be provided with the interval at the middle part of described double-edged blade and hold parts;

It is one that the interval of each described double-edged blade holds parts.

4. according to the cutter described in the claim 1-3, it is characterized in that:

Described interval holds parts and twines formation with adhesive tape.

5. according to arbitrary described cutter among the claim 1-4, it is characterized in that: the width in described gap is identical or different.

6. according to arbitrary described cutter among the claim 1-5, it is characterized in that: the width in described gap is identical; The width in described gap is 0.056-0.065cm; The width in described gap is specially 0.058cm.

7. according to arbitrary described cutter among the claim 1-6, it is characterized in that:

Described blade is 6.

8. the method for follicle before the mammal chamber of separate exsomatizing comprises the steps:

Draw from the cortex portion of the Mammalian Ovary that exsomatizes with arbitrary described cutter among the claim 1-7 and to get, obtain drawing and get thing, promptly obtain follicle before the chamber.

9. method according to claim 8 is characterized in that:

Described stroke of mode of getting got for drawing along horizontal and vertical both direction successively;

Before described stroke is got, also comprise the step of described stripped Mammalian Ovary being removed mesentery, corpus luteum and medullary substance;

Described stroke get after, comprise the steps: also described stroke got that thing once filters successively, secondary filter, the filtrate that obtains is centrifugal, get precipitation, obtains follicle before the chamber.

10. it is characterized in that according to Claim 8 or 9 described methods:

Filtering the described first time and adopting size is (180-220) μ m mesh screen, filters the described first time and adopts size to be specially 200 μ m mesh screens;

Filtering the described second time and adopting size is 80-120 μ m mesh screen, filters the described second time and adopts size to be specially 100 μ m mesh screens;

Described centrifugal rotation speed is 1450-1550r/min, and described centrifugal rotation speed is specially 1500r/min; The described centrifugal time is 4.5-5.5min, and the described centrifugal time is specially 5min, and described centrifugal radius of turn is 6.02cm;

Before each described filtration, also comprise and blow and beat the step of disperseing to be used for filtering mixture.

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