CN108342357A - A kind of high-purity leucocyte extracting method and device - Google Patents
A kind of high-purity leucocyte extracting method and device Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 23
- 238000000926 separation method Methods 0.000 claims abstract description 153
- 210000004369 blood Anatomy 0.000 claims abstract description 35
- 239000008280 blood Substances 0.000 claims abstract description 35
- 210000002381 plasma Anatomy 0.000 claims abstract description 29
- 238000004062 sedimentation Methods 0.000 claims abstract description 26
- 210000004027 cell Anatomy 0.000 claims abstract description 18
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 14
- 238000013517 stratification Methods 0.000 claims abstract description 12
- 238000005119 centrifugation Methods 0.000 claims abstract description 8
- 239000012141 concentrate Substances 0.000 claims abstract description 4
- 230000010148 water-pollination Effects 0.000 claims description 4
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 claims description 3
- 238000003780 insertion Methods 0.000 claims description 2
- 230000037431 insertion Effects 0.000 claims description 2
- 238000000605 extraction Methods 0.000 abstract description 5
- 239000004615 ingredient Substances 0.000 description 10
- 239000007788 liquid Substances 0.000 description 10
- 229920001612 Hydroxyethyl starch Polymers 0.000 description 9
- 229940050526 hydroxyethylstarch Drugs 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 230000005660 hydrophilic surface Effects 0.000 description 6
- 239000000654 additive Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 230000000996 additive effect Effects 0.000 description 4
- 230000005661 hydrophobic surface Effects 0.000 description 4
- 230000001788 irregular Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000002209 hydrophobic effect Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 241001391944 Commicarpus scandens Species 0.000 description 2
- 210000000601 blood cell Anatomy 0.000 description 2
- 235000019994 cava Nutrition 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000005436 troposphere Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
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Abstract
The present invention relates to leucocyte separation field, more particularly to a kind of high-purity leucocyte extracting method and device.A kind of high-purity leucocyte extraction element, including settling vessel, separation bag and stake body.Whole blood is first used settling vessel stratification, obtained rich leucocyte blood plasma to be transferred in separation bag by the present invention, and separation bag includes the thick intermediate thin three parts being sequentially communicated in both ends；Separation bag is fixed on stake body, is then centrifuged；After centrifugation, for enriched for leukemic cells at the middle part of separation bag, the bottom of separation bag, blood plasma described in erythrocyte sedimentation concentrate on the top of separation bag；Leucocyte in the middle part of separation bag is detached up to high-purity leucocyte.High-purity leucocyte extracting method provided by the invention and device, it is easy to operate, it is easy to implement, of low cost, isolated leucocyte purity is 97% or more, and high income.
Description
Technical field
The present invention relates to leucocyte separation fields, in particular to a kind of high-purity leucocyte extracting method and device.
Background technology
Currently, known common several leucocyte extraction elements and method are that blood is fitted into test tube or blood bag using ladder
It spends centrifugal process to obtain, the sedimentation agents such as hydroxyethyl starch is either added in blood and stands acquisition or hydroxyl second is added in blood
Gradient centrifugation obtains base starch again.
The past technical system is major problem is that the extraction purity of leucocyte and inefficient, main reasons is that haemocyte
After sedimentation separation, the leucocyte interface of formation is too thin, can not further be efficiently separated with red blood cell, causes operating procedure complicated,
The problems such as red blood cell is easily mixed into, and leucocyte is easy to be lost, or even have to use some cytotoxic additives, such as using leaching
The chemical substances such as bar cell separating liquid assist gradient centrifugation, need to be centrifuged repeatedly washing and remove the separating liquid, can be only achieved mitigation
Cytotoxicity, safe separating purpose.Therefore this also causes operating process time-consuming and laborious, leucocyte harvests quantity or quality not
It is high.In addition, also useful membrane filtration method harvests leucocyte, but because needing to rinse repeatedly, so ineffective.
In view of this, special propose the present invention.
Invention content
The first object of the present invention is to provide a kind of high-purity leucocyte extracting method, and this method first detaches whole blood
To rich leucocyte blood plasma, then rich leucocyte blood plasma is detached using separation bag provided by the invention again so that leucocyte
It is enriched in the elongated position in the middle part of separation bag, through detaching up to high-purity leucocyte.
The second object of the present invention is to provide the device for implementing the high-purity leucocyte extracting method, using the dress
It sets separation whole blood and obtains high-purity leucocyte, it is easy to operate, it is easy to implement, of low cost, separating effect is notable.
In order to realize that the above-mentioned purpose of the present invention, spy use following technical scheme：
A kind of high-purity leucocyte extracting method, includes the following steps：
By whole blood stratification, obtained rich leucocyte blood plasma is transferred in separation bag, and the separation bag includes that both ends are thick
The intermediate thin three parts being sequentially communicated；
The separation bag is fixed on stake body, is then centrifuged；
After centrifugation, enriched for leukemic cells is at the middle part of the separation bag, the bottom of separation bag, blood plasma described in erythrocyte sedimentation
Concentrate on the top of the separation bag；
Leucocyte in the middle part of the separation bag is detached up to the high-purity leucocyte.
A kind of high-purity leucocyte extracting method provided by the invention, first carries out initial gross separation by whole blood, obtains Fu Baixi
Born of the same parents' blood plasma；Obtained rich leucocyte blood plasma is detached through separation bag provided by the invention so that enriched for leukemic cells is in separation bag
Middle part, the middle part of separation bag is elongated portion, convenient for leucocyte and other compositions are separated, isolated leucocyte
For high-purity leucocyte.
The present invention also provides the devices for implementing above-mentioned high-purity leucocyte extracting method, including：Settling vessel, separation bag
And stake body；
Settling vessel：For by whole blood stratification；
Separation bag：For the isolated high-purity leucocyte of rich leucocyte blood plasma；
Stake body：For fixedly separated bag to be centrifuged.
Provided by the present invention for implementing the device of high-purity leucocyte extracting method, first whole blood is carried out with settling vessel
Layering, obtains rich leucocyte blood plasma；Rich leucocyte blood plasma is transferred to separation bag, and stake body centrifuges after fixing separation bag, different
Ingredient detaches, the enriched for leukemic cells part long and narrow in separation bag, isolated high-purity leucocyte.
Further, the settling vessel includes container, and the container is provided with entrance, the container with sedimentation side
It is unsymmetric structure to parallel adjacent inner wall.
Existing whole blood staticly settles device and is generally carried out in cylindrical container, such as the test tube of different size.But
Its sinking speed generally per hour 2mm, sinking speed is slow, and the yield after separation is bad.In consideration of it, general using centrifugation
Machine accelerated sedimentation, although CENTRIFUGAL ACCELERATING sedimentation, its yield and obtained material be still to be improved.In addition, using
Centrifuge also has cell itself certain damage, influences subsequently use or testing result, and increase lock out operation
Difficulty.
The present invention is unsymmetric structure by the way that the container adjacent inner wall parallel with sedimentation direction is arranged, and has upset whole blood cells
Stress balance situation in existing cylindrical settling vessel, improves the sinking speed and separating effect of each ingredient.Wherein,
It is unsymmetric structure that adjacent inner wall, which is between the unsymmetric structure i.e. adjacent plane of container inner wall, such as consistent with Blood Sedimentation direction
Section be non-disc, such as rectangular, polygon, arc, the asymmetrical section of irregular shape adjacent surface.For example, hold
The inner wall of device is four faces, is between the rectangle or two adjacent faces that between two adjacent faces are different in width
The face of trapezoidal etc. or other asymmetric irregular shapes of different in width；The inner wall of the application may be multiple faces, or
Person is the face of certain radian, and certainly, the radian of adjacent inner wall is different.
Inventor has found that the hydrophobicity of each plane of container is different, also influences the sinking speed and separating degree of cell.
Further, at least side of the container inner wall parallel with sedimentation direction has hydrophily.Due in blood
Ingredient to hydrophilic inner wall have bigger expansionary force, and for hydrophobic inner wall then with bigger convergent force, lead to
It crosses and inner wall is arranged to different hydrophobicitys, upset active force of the inner wall to cell, so that hanging down in a certain of settling vessel
Straight position forms the face that caves in so that and red blood cell drops to the bottom of settling vessel faster, and since reaction force is formed
Troposphere so that leucocyte is constantly increased, increase both separation speed and purity.
Whole blood is added into hydroxyethyl starch stratification relative to existing container, need 60min or so just and can make each layer it
Between demarcate apparent, and the yield of the ingredient detached only has 70%-80%；And settling vessel provided by the invention, by whole blood plus
Hydroxyethyl starch stratification, it is only necessary to the time of repose of 10-20min, you can each layer separation comes, after separation, obtains
Whole blood in leucocyte yield in 85%-98%.
Further, container can be cuboid or gengon, wherein being preferred with cuboid, it is preferable that the container
Shape be cuboid.The shape of container is cuboid both processing easy to produce, and reaches separation speed faster when standing
Spend and reach higher purity.Specifically, each length of side of cuboid, wherein the longest length of side be most short side length at least 2 times with
On.It can be such as 2 times, 3 times, 5 times, 8 times, 10 times, 15 times, not exceed 20 times generally.The size of specific container also needs
The volume of the whole blood to detach as needed determines.
In the present invention, the material of container is generally transparent or semitransparent plastics, preferably selects preferable with blood compatibility
Plastics, the medical polystyrene material used as existing.Its hydrophobic surface can keep its smooth with general technology, such as throw
Light, surface siliconization processing etc..
Using the method for above-mentioned settling vessel separating red corpuscle, include the following steps：
By in whole blood input pod, hydroxyethyl starch mixing is added, stands 10-20min, red blood cell is removed.
The additive amount of the additive amount of hydroxyethyl starch routinely carries out, and the quality for the hydroxyethyl starch generally added is dense
Degree is 5%-6%.
Then standing in step removes obtained rich leucocyte blood plasma generally using container is kept flat standing.
In addition, being stood using container provided by the invention, the time of standing does not exceed 30min, more than 30min
Afterwards, it floats on leucocyte above to be easy to break through the tunica albuginea layer of layering, into red blood cell layer, to reduce the yield of leucocyte.
Further, the separation bag includes the thick intermediate thin three parts being sequentially communicated in both ends, the separation bag it is outer
Edge is provided with overlap, and the overlap is equipped with suppending hole, and the separation bag is fixed on by the suppending hole on the stake body.
Preferably, the width at the middle part of the separation bag is less than the width of the bottom of the separation bag, the separation bag
The width of bottom is less than the width on the top of the separation bag.
Depending on content of the volume of the three parts of separation bag according to each ingredient of rich leucocyte blood plasma to be separated.Separation bag
Middle part width is most narrow, is fine strip shape, is for the ease of enriched for leukemic cells；And separation bag bottom width takes second place, and is for the ease of dividing
Go down from remaining erythrocyte sedimentation in the process；And the upper width of separation bag is most wide so that remaining blood plasma floats, Jiang Gecheng
Divide and preferably separates.
It is verified, it is detached using the separation bag of following shape, separating rate and separating degree are good.Further, institute
The top for stating separation bag is taper, and the middle part of the separation bag is vertical bar type, and the bottom of the separation bag is elliposoidal or circle.
Rich leucocyte blood plasma wall built-up in separation bag, separation are not thorough in order to prevent, it is preferable that the separation bag bag is internal
The part that wall connects is fillet, makes the inner edge continuous and derivable of bag body.
Entrance is arranged in the elongated part of separation bag, and being primarily to facilitate to be put into needs substance to be separated, and takes out separation
Good substance, inlet and outlet can be carried out by same pipeline.Further, the lower part at the middle part of the separation bag is provided with
Inlet tube.
Liquid to be separated is needed by inlet tube input, is drawn the liquid after separation by outlet, to prevent entrance
Manage the purity of liquid after remaining composition influence detaches.
It is highly preferred that the top at the middle part of the separation bag is provided with outlet.
Stake body plays the role of fixedly separated bag, i.e., separation bag trails, and then fixes, ensure bag body periphery by
Power is uniform.It is then placed in Centrifuge Cup to be centrifuged, in centrifugation, bag body does not fall off, does not tear, do not reveal.Further, described
The component for fixing the separation bag and be used to support the frame body that stake body includes frame body, is arranged on the frame body
Pedestal.
Further, the frame body is internally provided with the space mutually fitted with the separation bag so that after separation bag insertion
It is fixed.The space that separation bag size is mutually fitted is set to inside frame body, separation bag is embedded, and the suppending hole at separation bag edge passes through
It is fixed with the component of fixedly separated bag on frame body.Wherein, the component of fixedly separated bag can be on frame body：It slots on frame body
Afterwards, it is fixed with dismountable part, such as in frame body position trepanning corresponding with the suppending hole on separation bag overlap or slot, makes to hang
Hanging hole is fixed with frame body by screw thread or other component；Can also by frame body be arranged evagination hanging head be hung with
Reach fixed effect.
In addition, the clamping plate that can be adjusted can be also arranged for adjusting in the top and bottom of the holder vertical with Centrifuge Cup section
The capacity of the bag body of separation bag, and clamping device is set to control the discrepancy of liquid in two ports that come in and go out of bag body.
Compared with prior art, beneficial effects of the present invention are：
(1) high-purity leucocyte extracting method provided by the invention, this method is first by the isolated rich leucocyte blood of whole blood
Slurry, is then again detached rich leucocyte blood plasma using separation bag provided by the invention so that enriched for leukemic cells is in separation bag
The elongated position at middle part, through detaching the high-purity leucocyte for being 97% or more up to purity.
(2) high-purity leucocyte extraction element provided by the invention, it is easy to operate, it is easy to implement, of low cost, separation effect
Fruit is notable.
(3) settling vessel provided by the invention, time of repose only need 10-20min, efficiently quickly float on leucocyte
The yield on the top of red blood cell, the leucocyte being isolated to reaches 85% or more, and the purity of leucocyte reaches 90% or more.
(4) thin both sides are thick among separation bag provided by the invention, and after centrifugation, obtained enriched for leukemic cells is narrow in separation bag
Long part, remaining ingredient are then distributed in the both sides of separation bag, not only 95% or more isolated leucocyte yield, and pure
Degree is 97% or more, and separating effect is notable.
(5) the present invention also provides the stake bodies of fixed separation bag, ensure separation bag periphery uniform force, are centrifuging
When, bag body does not fall off, does not tear, does not reveal.
Description of the drawings
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art are briefly described, it should be apparent that, in being described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, other drawings may also be obtained based on these drawings.
Fig. 1 is the stereogram of settling vessel provided in an embodiment of the present invention；
Fig. 2 is the front view of settling vessel shown in FIG. 1；
Fig. 3 is the structural schematic diagram of separation bag provided in an embodiment of the present invention；
Fig. 4 is the vertical view of stake body provided in an embodiment of the present invention；
Fig. 5 is the front view of stake body provided in an embodiment of the present invention；
Reference numeral：
1- containers；2- entrances；11- hydrophilic surfaces；3- separation bags；The top of 31- separation bags；The middle part of 32- separation bags；
The bottom of 33- separation bags；4- overlaps；5- suppending holes；6- outlets；7- inlet tubes；8- frame bodies；9- pedestals；Inside 10- frame bodies
Space.
Specific implementation mode
Technical scheme of the present invention is clearly and completely described below in conjunction with attached drawing, it is clear that described implementation
Example is a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, ordinary skill
The every other embodiment that personnel are obtained without making creative work, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that term "center", "upper", "lower", "left", "right", "vertical",
The orientation or positional relationship of the instructions such as "horizontal", "inner", "outside" be based on the orientation or positional relationship shown in the drawings, merely to
Convenient for the description present invention and simplify description, do not indicate or imply the indicated device or element must have a particular orientation,
With specific azimuth configuration and operation, therefore it is not considered as limiting the invention.In addition, term " first ", " second ",
" third " is used for description purposes only, and is not understood to indicate or imply relative importance.
In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " installation ", " phase
Even ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or be integrally connected；It can
Can also be electrical connection to be mechanical connection；It can be directly connected, can also indirectly connected through an intermediary, Ke Yishi
Connection inside two elements.For the ordinary skill in the art, above-mentioned term can be understood at this with concrete condition
Concrete meaning in invention.
The device of high-purity leucocyte extraction provided by the invention, including：Settling vessel, separation bag and stake body；
Settling vessel：For by whole blood stratification；
Separation bag：Isolated high-purity leucocyte for rich leucocyte blood plasma；
Stake body：For fixedly separated bag to be centrifuged.
Wherein, existing container may be used in settling vessel, can also use settling vessel provided by the invention.It stands and holds
For device by whole blood stratification, the content of obtained rich leucocyte blood plasma influences the yield of follow-up high-purity leucocyte.
Fig. 1 is the top perspective view of settling vessel provided in an embodiment of the present invention；Fig. 2 is settling vessel shown in FIG. 1
Front view.As shown in Figs. 1-2, settling vessel provided in an embodiment of the present invention, including container 1, the container 1 are provided with entrance
2, the adjacent inner wall parallel with sedimentation direction of the container 1 is unsymmetric structure.
The present invention is unsymmetric structure by the way that the container adjacent inner wall parallel with sedimentation direction is arranged, and has upset whole blood cells
Stress balance situation in existing cylindrical settling vessel, improves the sinking speed and separating effect of each ingredient.
Specifically, it is generally carried out in cylindrical container relative to existing stood for whole blood, such as different size
Test tube, sedimentation need 1h or so, and the yield after separation is below 80%；And centrifuge accelerated sedimentation is used, to cell itself
Also there is certain damage, influence subsequently use or testing result, and increase the difficulty of lock out operation.It is provided by the invention red
Cell standing sedimentation separator, sedimentation only need 10-20min, and the yield after separation is 85% or more.
Wherein, it is non-that container adjacent inner wall provided by the invention, which is between the unsymmetric structure i.e. adjacent plane of container inner wall,
Symmetrical structure, such as section consistent with Blood Sedimentation direction are non-disc, such as rectangular, polygon, arc, irregular shape be adjacent
The asymmetrical section in surface.For example, the inner wall of container is four faces, is the rectangular of different in width between two adjacent faces
Shape, for the trapezoidal etc. of different in width or other are asymmetric as depicted in figs. 1 and 2 or between two adjacent faces
The face of irregular shape；The inner wall of the application may be multiple faces, or be the face of certain radian, certainly, the arc of adjacent inner wall
Degree is different.
Inventor has found that the hydrophobicity of each plane of container is different, also influences the sinking speed and separating degree of cell.
Further, at least side of the container inner wall parallel with sedimentation direction has hydrophily.As shown in Figure 2
Thick line be expressed as hydrophilic surface 3, the line not blackened is then considered as hydrophobic surface.The container at least side inner wall parallel with sedimentation direction
For hydrophilic surface 3, by sedimentation telegoniometer, container inner wall in parallel can be hydrophilic surface 3 there are one face, remaining is hydrophobic
Face；Can also be hydrophilic surface 3 there are two inner wall, remaining is hydrophobic surface；Can also be hydrophilic surface 3 there are three inner wall, remaining
For hydrophobic surface；Etc., be just not listed one by one.
Since the ingredient in blood has hydrophilic inner wall the expansionary force of bigger, and then have for hydrophobic inner wall
The convergent force for having bigger upsets active force of the inner wall to cell by the way that inner wall to be arranged to different hydrophily and hydrophobicity, from
And to form the face that caves in a certain upright position of settling vessel so that red blood cell drops to settling vessel faster
Bottom, and since the convection action that reaction force is formed makes leucocyte constantly rise, increase the speed and purity of the two separation, obtains
To leucocyte blood plasma in leucocyte yield be 85% or more.
Whole blood is added into hydroxyethyl starch stratification relative to existing container, need 60min or so just and can make each layer it
Between demarcate apparent, and the yield of the ingredient detached only has 60%-80%；And settling vessel provided by the invention, by whole blood plus
Hydroxyethyl starch stratification, it is only necessary to the time of repose of 10-20min, you can each layer separation is come, separating effect such as Fig. 3
Shown, after separation, the yield of the leucocyte in obtained whole blood is 85% or more.
Wherein, the additive amount of the additive amount of hydroxyethyl starch routinely carries out.
In addition, carrying out stratification to whole blood using settling vessel provided in an embodiment of the present invention, the time of standing should not
More than 30min, after 30min, floats on leucocyte above and be easy to break through the tunica albuginea layer of layering, into red blood cell layer.
Further, container can be that cuboid or gengon are preferred wherein with cuboid, it is preferable that the container
Shape be cuboid, as shown in Figure 1.The shape of container is cuboid both processing easy to produce, and is reached more when standing
Fast separating rate and reach higher purity.Specifically, each length of side of cuboid, wherein the longest length of side is most short side length
At least 2 times or more.It can be such as 2 times, 3 times, 5 times, 8 times, 10 times, 15 times, not exceed 20 times generally.Specific container
Size also needs to the volume of the whole blood detached as needed to determine.
Fig. 3 is the structural schematic diagram of separation bag provided in an embodiment of the present invention；Fig. 4 and Fig. 5 is the structural representation of stake body
Figure.
As shown in figure 3, separation bag includes the thick intermediate thin three parts being sequentially communicated in both ends, the outer rim setting of separation bag 3
There are overlap 4, the overlap 4 to be equipped with suppending hole 5, the separation bag 3 is fixed on by the suppending hole 5 on the stake body.
Further, the width at the middle part 32 of separation bag is less than the width of the bottom 33 of separation bag, the bottom 33 of separation bag
Width be less than separation bag top 31 width.
Depending on content of the volume of the three parts of separation bag according to each ingredient of rich leucocyte blood plasma to be separated.Separation bag
Middle part width is most narrow, is fine strip shape, is for the ease of enriched for leukemic cells；And separation bag bottom width takes second place, and is for the ease of dividing
Go down from remaining erythrocyte sedimentation in the process；And the upper width of separation bag is most wide so that remaining blood plasma floats, Jiang Gecheng
Divide and preferably separates.
It is verified, it is detached using the separation bag 3 of following shape, separating rate and separating degree are good.Further, institute
The top 31 for stating separation bag is taper, and the middle part 32 of the separation bag is vertical bar type, and the bottom 33 of the separation bag is elliposoidal
Or it is round.
Rich leucocyte blood plasma wall built-up, separation in separation bag 3 are not thorough in order to prevent, it is preferable that the separation bag bag body
The part that inner wall connects is fillet, makes the inner edge continuous and derivable of bag body.
Entrance is arranged in the elongated part of separation bag 3, and being primarily to facilitate to be put into needs substance to be separated, and takes out separation
Good substance, inlet and outlet can be carried out by same pipeline.Further, the lower part setting at the middle part 32 of the separation bag
There is inlet tube 7.
Need liquid to be separated by the input of inlet tube 7, drawn the liquid after separation by outlet 6, with prevent into
Mouth manages the purity of liquid after remaining composition influence separation.
It is highly preferred that the top at the middle part 32 of the separation bag is provided with outlet 6.
Stake body as shown in Figures 4 and 5 plays the role of fixedly separated bag, i.e., separation bag trails, and then fixed
Firmly, ensure bag body periphery uniform force.The separation bag fixed and stake body are put into togerther in the Centrifuge Cup of centrifuge, from
When the heart, it is ensured that bag body does not fall off, does not tear, do not reveal.
Further, the stake body include frame body 8, be arranged on the frame body for fixing the portion of the separation bag
Part and the pedestal 9 for being used to support the frame body.
Further, the frame body is internally provided with the space fitted with 3 phase of the separation bag so that the separation bag 3 is embedded in
After fix.Space 10 inside frame body is mutually fitted with 3 size of separation bag, and separation bag 3 is embedded, the suppending hole 5 at 3 edge of separation bag
By being fixed with the component of fixedly separated bag on frame body 8.Wherein, the component of fixedly separated bag can be on frame body：On frame body
After fluting, fixed with dismountable part, such as in frame body position trepanning corresponding with the suppending hole 5 on 3 overlap 4 of separation bag or
Slot, makes that suppending hole passes through screw thread with frame body or other component is fixed；Can also by frame body 8 be arranged evagination hanging head into
Row suspension is to reach fixed effect.
In addition, the clamping plate that can be adjusted can be also arranged for adjusting in the top and bottom of the frame body 8 vertical with Centrifuge Cup section
The capacity of the bag body of separation bag 3 is saved, and clamping device is set to control the discrepancy of liquid in two ports that come in and go out of bag body.
Specifically, settling vessel carries out initial gross separation to whole blood, obtains rich leucocyte blood plasma；Obtained rich leucocyte blood plasma
Input separation bag 3 is detached so that for enriched for leukemic cells at the middle part of separation bag 32, the middle part 32 of separation bag is elongated portion；
The bottom 33 of separation bag described in erythrocyte sedimentation, blood plasma concentrate on the top 31 of the separation bag；By leucocyte and other compositions
It separates, isolated leucocyte is high-purity leucocyte, and purity reaches 97% or more, is come relative to rich leucocyte blood plasma
It says, the yield of obtained leucocyte reaches 95% or more.
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Present invention has been described in detail with reference to the aforementioned embodiments for pipe, it will be understood by those of ordinary skill in the art that：Its according to
So can with technical scheme described in the above embodiments is modified, either to which part or all technical features into
Row equivalent replacement；And these modifications or replacements, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of high-purity leucocyte extracting method, which is characterized in that include the following steps：
By whole blood stratification, obtained rich leucocyte blood plasma is transferred in separation bag, and the separation bag includes that both ends are slightly intermediate
The thin three parts being sequentially communicated；
The separation bag is fixed on stake body, is then centrifuged；
After centrifugation, enriched for leukemic cells at the middle part of the separation bag, concentrate by the bottom of separation bag described in erythrocyte sedimentation, blood plasma
On the top of the separation bag；
Leucocyte in the middle part of the separation bag is detached up to the high-purity leucocyte.
2. implementing a kind of device of high-purity leucocyte extracting method described in claim 1, which is characterized in that including：It stands
Container, separation bag and stake body；
Settling vessel：For by whole blood stratification；
Separation bag：Isolated high-purity leucocyte for rich leucocyte blood plasma；
Stake body：For fixedly separated bag to be centrifuged.
3. the apparatus of claim 2, which is characterized in that the settling vessel includes container, and the container is provided with
The adjacent inner wall parallel with sedimentation direction of entrance, the container is unsymmetric structure；
Preferably, at least side of the container inner wall parallel with sedimentation direction has hydrophily；
The container is rectangular parallelepiped structure.
4. the apparatus of claim 2, which is characterized in that the separation bag, which includes that both ends are thick intermediate thin, to be sequentially communicated
Three parts, the outer rim of the separation bag is provided with overlap, and the overlap is equipped with suppending hole, and the separation bag passes through described outstanding
Hanging hole is fixed on the stake body.
5. device according to claim 4, which is characterized in that the width at the middle part of the separation bag is less than the separation bag
Bottom width, the width of the bottom of the separation bag is less than the width on the top of the separation bag.
6. device according to claim 4, which is characterized in that the top of the separation bag is taper, the separation bag
Middle part is vertical bar type, and the bottom of the separation bag is elliposoidal or circle.
7. device according to claim 4, which is characterized in that the part that the internal wall of separation bag bag connects is circle
Angle.
8. device according to claim 4, which is characterized in that the lower part at the middle part of the separation bag is provided with inlet tube；
Preferably, the top at the middle part of the separation bag is provided with outlet.
9. the apparatus of claim 2, which is characterized in that the stake body includes frame body, is arranged on the frame body
For fixing the component of the separation bag and being used to support the pedestal of the frame body.
10. device according to claim 9, which is characterized in that the frame body is internally provided with mutually fits with the separation bag
Space so that fixed after separation bag insertion.
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| CN201810431306.0A CN108342357B (en) | 2018-05-08 | 2018-05-08 | High-purity leukocyte extraction method and device |
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| CN108498329A (en) * | 2018-05-09 | 2018-09-07 | 深圳市祥平元创科技有限公司 | A kind of blood bag |
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| CN108342357B (en) | 2024-04-05 |
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