CN115141800A - Autoblood treatment method and autoblood treatment device - Google Patents
Autoblood treatment method and autoblood treatment device Download PDFInfo
- Publication number
- CN115141800A CN115141800A CN202210832461.XA CN202210832461A CN115141800A CN 115141800 A CN115141800 A CN 115141800A CN 202210832461 A CN202210832461 A CN 202210832461A CN 115141800 A CN115141800 A CN 115141800A
- Authority
- CN
- China
- Prior art keywords
- filter membrane
- flexible
- array filter
- red blood
- module
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 74
- 239000012528 membrane Substances 0.000 claims abstract description 207
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 165
- 210000004369 blood Anatomy 0.000 claims abstract description 131
- 239000008280 blood Substances 0.000 claims abstract description 131
- 210000000265 leukocyte Anatomy 0.000 claims abstract description 70
- 238000001914 filtration Methods 0.000 claims abstract description 68
- 238000004140 cleaning Methods 0.000 claims description 50
- 239000011148 porous material Substances 0.000 claims description 50
- 238000005406 washing Methods 0.000 claims description 24
- 230000000694 effects Effects 0.000 claims description 19
- 239000007924 injection Substances 0.000 claims description 15
- 238000002347 injection Methods 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 15
- 239000000523 sample Substances 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 230000008595 infiltration Effects 0.000 claims description 12
- 238000001764 infiltration Methods 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 10
- 238000003860 storage Methods 0.000 claims description 9
- 230000009471 action Effects 0.000 claims description 7
- 238000007865 diluting Methods 0.000 claims description 7
- 238000011156 evaluation Methods 0.000 claims description 7
- 239000008055 phosphate buffer solution Substances 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 6
- 238000004043 dyeing Methods 0.000 claims description 4
- 239000012723 sample buffer Substances 0.000 claims description 3
- 239000002504 physiological saline solution Substances 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 abstract description 19
- 210000002381 plasma Anatomy 0.000 abstract description 6
- 230000006378 damage Effects 0.000 abstract description 5
- 238000011084 recovery Methods 0.000 description 11
- 230000008569 process Effects 0.000 description 8
- 238000010186 staining Methods 0.000 description 5
- 239000003344 environmental pollutant Substances 0.000 description 4
- 231100000719 pollutant Toxicity 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 210000003855 cell nucleus Anatomy 0.000 description 3
- 210000000805 cytoplasm Anatomy 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000003672 processing method Methods 0.000 description 3
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 2
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000002745 absorbent Effects 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 239000003114 blood coagulation factor Substances 0.000 description 2
- 229940019700 blood coagulation factors Drugs 0.000 description 2
- 230000005779 cell damage Effects 0.000 description 2
- 208000037887 cell injury Diseases 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000006073 displacement reaction Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000004907 flux Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 235000001637 Ganoderma lucidum Nutrition 0.000 description 1
- 240000008397 Ganoderma lucidum Species 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 241001317416 Lius Species 0.000 description 1
- 206010071229 Procedural haemorrhage Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 208000003441 Transfusion reaction Diseases 0.000 description 1
- 206010053476 Traumatic haemorrhage Diseases 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000006727 cell loss Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0081—Purging biological preparations of unwanted cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/08—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by vibration
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/12—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by pressure
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0641—Erythrocytes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
- C12N2509/10—Mechanical dissociation
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Sustainable Development (AREA)
- Cell Biology (AREA)
- Hematology (AREA)
- Immunology (AREA)
- External Artificial Organs (AREA)
Abstract
The invention provides a method and a device for treating autologous blood, wherein the sizes of a first flexible micropore array filter membrane and a second flexible micropore array filter membrane in the embodiment of the invention can be accurately defined, the first flexible micropore array filter membrane can be used for effectively removing white blood cells in the autologous blood based on size difference, the second flexible micropore array filter membrane is used for effectively replacing solution components in the filtered blood, the red blood cells are cleaned, and finally the cleaned red blood cells are recovered, so that the white blood cells/blood plasma in the autologous blood can be efficiently removed. The flexible microporous array filter membrane in the embodiment of the invention has the thickness of micron level (not more than 10 microns), the cell stroke is obviously shortened during filtration, and the possibility of damage to red blood cells can be effectively reduced. The embodiment of the invention also lays a net-shaped supporting structure below the flexible micropore array filter membrane, thereby improving the filtration strength which can be borne by the flexible micropore array filter membrane on the premise of not changing the microstructure of the flexible micropore array filter membrane.
Description
Technical Field
The invention relates to the field of biomedical basic research and clinical medicine, in particular to a method and a device for treating autoblood.
Background
Autologous blood transfusion means that intraoperative/traumatic bleeding is collected and timely transfused to a patient, and the collected blood is filtered and cleaned under the condition that the collected blood possibly contains pollutants such as platelets, activated blood coagulation factors, high-concentration potassium ions, free hemoglobin, anticoagulation and the like.
At present, common autoblood instruments realize separation of red blood cells and pollutants based on a centrifugal principle, and the methods have high treatment efficiency but have inherent defects, wherein the centrifugal process inevitably damages the cells and influences the activity of the cells; on the other hand, the centrifugal operation determines that the instrument is relatively bulky and is not suitable for use in a severe environment in a battlefield, away from a medical facility, or the like.
In order to overcome the negative effects brought by centrifugation, a method based on filter membrane filtration is provided, the recovery of blood in the operation is realized mainly based on a microporous filter membrane and a super absorbent material membrane, the collected blood flows through an upper layer blood bag during working, and the components with larger sizes, such as red blood cells, white blood cells and the like in the blood cannot pass through the microporous filter membrane and enter an output flow path; and pollutants with smaller sizes in the blood can pass through the microporous filter membrane and be adsorbed by the super absorbent material wrapped by the microporous filter membrane. However, this method cannot remove white blood cells larger in size than red blood cells, easily causes transfusion reaction, and also causes damage to red blood cells due to high-speed blood flow during filtration.
It follows that in the related art, the method based on the centrifugal principle has high processing efficiency but large volume; the method based on the filter membrane filtration is small in volume and portable, but the treatment efficiency is limited; in addition, it is difficult to secure the activity of erythrocytes in both of these methods. A new portable and efficient autologous blood treatment method is needed to realize efficient removal of leukocytes/plasma and high-activity recovery of erythrocytes in autologous blood under a severe environment.
Disclosure of Invention
In order to solve the problems in the background art, the invention provides a method and a device for treating autologous blood, so as to realize high-efficiency removal of white blood cells/blood plasma and high-activity recovery of red blood cells in the autologous blood.
In a first aspect, the present invention provides a method of autologous blood treatment, the method comprising:
autologous blood collection step: collecting blood of a user and storing the blood into a first container;
transferring the autologous blood from the first container to a sample buffer container in a leukocyte removal module;
a leukocyte filtration step: under the effect of external force that vortex oscillator was applyed, the first flexible micropore array filter membrane in the module is got rid of to leucocyte is right autologous blood filters, collects and filters back blood, deposits in the second container, wherein, external force orthogonal in the flow direction of the fluid in the module is got rid of to leucocyte, first flexible micropore array filter membrane is used for the filtering leucocyte in the autologous blood, the filtration pore diameter of first flexible micropore array filter membrane is 6-10 microns, and the filtration pore spacing is 2-4 microns, and thickness is not more than 10 microns, first netted bearing structure has been laid under the first flexible micropore array filter membrane, first netted bearing structure is used for supporting first flexible micropore array filter membrane, first netted bearing structure comprises a plurality of first netted supporting unit, a plurality of first netted supporting unit evenly distributed in first flexible micropore array filter membrane below, the aperture of first netted supporting unit is greater than the filtration pore diameter of first flexible micropore array filter membrane, the area of first supporting unit is less than 130 mm's filtration pore diameter, first flexible micropore array filter membrane's area is less than 130mm 2 。
Optionally, the method further comprises:
transferring the filtered blood from the second container to a sample buffer container in a red blood cell washing module;
and (3) red blood cell cleaning: under the effect of the external force that vortex oscillator applyed, to last add red blood cell washing liquid in the red blood cell washing module, utilize flexible micropore array filter membrane of second in the red blood cell washing module is right blood after filtering washs, wherein, external force orthogonal in the flow direction of the fluid in the red blood cell washing module, the flexible micropore array filter membrane of second is used for catching red blood cells, the filtration pore diameter of the flexible micropore array filter membrane of second is 2-4 microns, and the filtration pore interval is 2 microns, and thickness is not more than 10 microns, the netted bearing structure of second has been laid under the flexible micropore array filter membrane of second, the netted bearing structure of second is used for supporting the flexible micropore array filter membrane of second, the netted bearing structure of second comprises the netted bearing unit of a plurality of secondThe plurality of second reticular supporting units are uniformly distributed below the second flexible micropore array filter membrane, the aperture of each second reticular supporting unit is larger than the aperture of the filter pore of the second flexible micropore array filter membrane, and the area of each second reticular supporting unit is smaller than 25mm 2 ；
The liquid outlet side of the red blood cell cleaning module is connected with a negative pressure module, and the red blood cell cleaning module is provided with a red blood cell collecting module for storing the treated red blood cells;
and (3) red blood cell collection: and applying an external force orthogonal to the plane of the second flexible microporous array filter membrane to the red blood cell cleaning module by using the negative pressure module, so that the red blood cells pass through the filter pores of the second flexible microporous array filter membrane and reach the red blood cell collecting module, and the red blood cells captured on the second flexible microporous array filter membrane are collected to the red blood cell collecting module.
Optionally, the negative pressure module is an injection pump, and the liquid outlet side of the red blood cell cleaning module is connected to the negative pressure module and provided with a red blood cell collecting module, including:
the liquid outlet side of the red blood cell cleaning module is connected with an injection pump, the injection pump comprises an injector, and the injector is used for temporarily storing red blood cells;
the red blood cell collecting step comprises: under the action of an external force applied by the injection pump and orthogonal to the plane of the second flexible microporous array filter membrane, the red blood cells pass through the filter pores of the second flexible microporous array filter membrane and are temporarily stored in an injector connected with the injection pump, and then the red blood cells are transferred to the red blood cell collection module from the injector, so that the red blood cells captured on the second flexible microporous array filter membrane are collected to the red blood cell collection module.
Optionally, the shape of the first mesh supporting unit or the shape of the second mesh supporting unit is: any one of regular hexagon, circle, regular quadrangle and regular trilateral;
the aperture shape of the first net-shaped supporting unit or the aperture shape of the second net-shaped supporting unit is as follows: any one of regular hexagon, circle, regular quadrangle and regular trilateral.
Optionally, after the red blood cell collecting step, the method further comprises:
evaluation step of autoblood treatment effect: and taking out the second flexible micropore array filter membrane from the erythrocyte cleaning module, dyeing and counting leukocytes of the second flexible micropore array filter membrane, counting the number of the leukocytes captured by the second flexible micropore array filter membrane, judging whether the number of the leukocytes is smaller than a preset value or not, and obtaining an evaluation result.
Optionally, prior to the leukocyte filtering step, the method further comprises: a first flexible microporous array filter membrane treatment step: performing infiltration treatment on the first flexible microporous array filter membrane by using alcohol, and after the infiltration treatment, diluting and removing the alcohol attached to the first flexible microporous array filter membrane by using a phosphate buffer solution;
prior to the red blood cell washing step, the method further comprises:
a second flexible microporous array filter membrane treatment step: and performing infiltration treatment on the second flexible microporous array filter membrane by using alcohol, and after the infiltration treatment, diluting and removing the alcohol attached to the second flexible microporous array filter membrane by using a phosphate buffer solution.
Optionally, the operating conditions of the vortex oscillator include: the rotation speed is 200-3000rpm, and the vortex oscillator is periodically operated at 30s intervals.
Optionally, in the red blood cell washing step, the washing solution is normal saline, and the volume of the washing solution is 1/2 of the volume of the autologous blood.
In a second aspect, the present invention provides an autologous blood treatment device, the device comprising: the device comprises a first container, a white blood cell removing module, a second container, a red blood cell cleaning module, a vortex oscillator, a negative pressure module and a red blood cell collecting module;
the device is used for executing the autologous blood treatment method of the first aspect.
Optionally, a first leukocyte removal module is disposed in the leukocyte removal moduleNetted bearing structure, first netted bearing structure is used for supporting first flexible micropore array filter membrane, first netted bearing structure comprises a plurality of first netted supporting unit, a plurality of first netted supporting unit evenly distributed in first flexible micropore array filter membrane below, the aperture of first netted supporting unit is greater than the filtration pore aperture of first flexible micropore array filter membrane, the area of first netted supporting unit is less than 130mm 2 ；
The second netted bearing structure is used for supporting the flexible micropore array filter membrane of second, the netted bearing structure of second comprises the netted supporting element of a plurality of second, a plurality of netted supporting element evenly distributed of second are in the flexible micropore array filter membrane below of second, the aperture of the netted supporting element of second is greater than the filtration pore aperture of the flexible micropore array filter membrane of second, the area of the netted supporting element of second is less than 25mm 2 。
Compared with the prior art, the invention has the following advantages:
(1) The embodiment of the invention realizes the highly effective autologous blood treatment. The sizes of the first flexible micropore array filter membrane and the second flexible micropore array filter membrane in the embodiment of the invention can be accurately defined, and the first flexible micropore array filter membrane can be used based on size difference to apply external force in the direction parallel to the plane of the filter membrane, so that leukocytes in autologous blood can be effectively removed; by combining with external force applied in the direction parallel to the plane of the filter membrane, the solution component in the filtered blood is effectively replaced by using the second flexible microporous array filter membrane, so that the red blood cells are cleaned; and finally, recovering the cleaned red blood cells by combining with an external force applied in the direction orthogonal to the plane of the filter membrane, thereby realizing the high-efficiency removal of the white blood cells/blood plasma in the autologous blood. In addition, the leukocyte removal efficiency can be fully sampled and evaluated in a mode of dyeing and counting through the second flexible micropore array filter, and the leukocyte removal effectiveness is further guaranteed.
(2) Realizes the autoblood treatment with high activity. The flexible microporous array filter membrane in the embodiment of the invention has the thickness of micron level (not more than 10 microns), the cell stroke is obviously shortened during filtration, and the possibility of damage to red blood cells can be effectively reduced. Considering that the strength of the flexible microporous array filter membrane is influenced due to the small thickness of the flexible microporous array filter membrane; in addition, in order to increase the volume capable of treating autologous blood, the area of the flexible microporous array filter membrane also needs to be increased correspondingly, and the possibility of filter failure caused by filter membrane breakage is further increased. Therefore, the embodiment of the invention also lays the reticular support structure below the flexible micropore array filter membrane, thereby improving the filtration strength which can be borne by the flexible micropore array filter membrane on the premise of increasing the filtration area and keeping the thickness of the filter membrane unchanged. In the embodiment of the invention, the net-shaped supporting structure is composed of a plurality of net-shaped supporting units, and the net-shaped supporting units are independent of the flexible microporous array filter membrane, so that compared with a method for designing a supporting structure in the filter membrane, the supporting strength is improved, and meanwhile, the production difficulty and cost are obviously reduced.
Drawings
FIG. 1 is a flow chart of a method of autologous blood treatment according to an embodiment of the present invention;
FIG. 2 is a flow chart of another method of autologous blood treatment in an embodiment of the present invention;
FIG. 3 is a schematic illustration of an autologous blood treatment device in an embodiment of the present invention;
FIG. 4 is a schematic diagram showing the comparison of the cell staining results of the first flexible microporous array filter membrane and the second flexible microporous array filter membrane after being processed by an autologous blood processing method according to the embodiment of the present invention.
Detailed Description
The following examples are provided to better understand the present invention, not to limit the best mode, and not to limit the content and protection scope of the present invention, and any product that is the same or similar to the present invention and is obtained by combining the present invention with other features of the prior art and the present invention falls within the protection scope of the present invention.
The examples do not indicate specific experimental procedures or conditions, and can be performed according to the procedures or conditions of the conventional experimental procedures described in the literature in the field. The reagents and other instruments used are not indicated by manufacturers, and are conventional reagent products which can be purchased in the market.
In a first aspect, an embodiment of the present invention provides an autologous blood treatment method, and fig. 1 is a flowchart of an autologous blood treatment method in an embodiment of the present invention; as shown in fig. 1, the method comprises the steps of:
s101, autologous blood collection: and collecting autologous blood of the user and storing the autologous blood into the first container.
In practice, a medical sterile instrument (such as a spoon or a syringe) can be used to collect and store the bleeding of the patient in the first container.
And S102, transferring the autologous blood from the first container to a sample temporary storage container in the leukocyte removal module.
In one embodiment, the first container may be connected to the upper portion of the leukoreduction module, so that the autologous blood in the first container flows into the temporary sample storage container of the leukoreduction module at a uniform speed.
S103, leukocyte filtration step: under the action of external force applied by the vortex oscillator, the autologous blood is filtered by the first flexible micropore array filter membrane in the leukocyte removal module, and the filtered blood is collected and stored in the second container.
The external force is orthogonal to the flow direction of fluid in the leukocyte removal module, the first flexible micropore array filter membrane is used for filtering leukocytes in the autologous blood, the pore size of filter pores of the first flexible micropore array filter membrane is 6-10 micrometers, the space between the filter pores is 2-4 micrometers, and the thickness is not more than 10 micrometers; first netted bearing structure has been laid under first flexible micropore array filter membrane, first netted bearing structure is used for supporting first flexible micropore array filter membrane, first netted bearing structure comprises a plurality of first netted support unit, a plurality of first netted support unit evenly distributed in first flexible micropore array filter membrane below, the aperture of first netted support unit is greater than the filtration pore aperture of first micropore array filter membrane, the area of first netted support unit is less than 130mm 2 。
In the embodiment of the invention, considering that the white blood cells are generally spherical cells with the diameter of 6-20 microns, the red blood cells are round cake-shaped cells with the diameter of about 6 microns and the thickness of about 2 microns, theoretically, when the size of the filter hole of the filter membrane is 6 microns, the white blood cells in the autologous blood can be removed on the premise of not damaging the red blood cells, and the main components in the filtered blood are the red blood cells and other components in the plasma. Although the smaller the filter pore size of the filter membrane (close to 6 microns), the higher the efficiency of leukocyte removal from autologous blood, the longer the filtration time and the greater the damage to erythrocytes. Considering that the leukocyte size distribution conforms to the positive distribution, the lower limit is about 6 microns, and the increase of the filter pore size of the filter membrane does not cause the significant reduction of the leukocyte removal efficiency, in practical application, the filter pore diameter of the first flexible micropore array filter membrane can be 6-10 microns. In specific application, a first flexible microporous array filter membrane with a proper pore size can be selected from 6-10 micrometers according to actual requirements.
In the embodiment of the invention, it is also considered that the larger the filter pore spacing of the filter membrane is, the higher the probability that non-target cells with the size smaller than the filter pore diameter fall between the filter pores is, and the lower the filtering effectiveness is, so the smaller the filter pore spacing is, the higher the filtering effectiveness is, but the smaller the filter pore spacing is, the lower the overall strength of the filter membrane is, and after comprehensive consideration is given, the filter pore spacing is selected to be 2-4 micrometers.
In the embodiment of the invention, in order to reduce the cell stroke of red blood cells in the filtering process and reduce red blood cell damage, the invention provides a flexible microporous filter membrane which is obtained by making the thickness of the filter membrane of the microporous array filter membrane not more than 10 microns and is used for the autologous blood treatment method provided by the embodiment of the invention.
In the embodiment of the invention, the area of the first flexible microporous array filter membrane can be determined according to the volume of autologous blood which needs to be treated actually. In the embodiment of the invention, under the condition that the area of the filter membrane is large enough, a large-volume autologous blood sample can be quickly processed.
In the embodiment of the invention, the overall strength of the first flexible micropore array filter membrane is considered to be smaller, and in the application process, the mesh-shaped support structure is laid under the first flexible micropore array filter membrane, so that the strength which can be borne by the flexible micropore array filter membrane is improved on the premise of not changing the microstructure of the flexible micropore array filter membrane.
In the embodiment of the invention, the net-shaped supporting structure is formed by a plurality of net-shaped supporting units, so that the supporting strength of the net-shaped supporting structure can be flexibly adjusted by adjusting the number and distribution of the net-shaped supporting units, and the aim of adjusting the strength of the filter membrane is further fulfilled. Therefore, in practical application, the strength requirement of the filter membrane can be determined according to actual needs, and a proper number of reticular support units are selected to form the reticular support structure. For example, when the requirement for the strength of the filter membrane is low when the volume of the autologous blood to be treated is small, a smaller number of the mesh-shaped supporting units can be selected to form the mesh-shaped supporting structure, and when the requirement for the strength of the filter membrane is high when the volume of the autologous blood to be treated is large, a larger number of the mesh-shaped supporting units can be selected to form the mesh-shaped supporting structure.
In the embodiment of the invention, considering that the net-shaped supporting structure is obtained by splicing a plurality of net-shaped supporting units, compared with the whole net-shaped supporting structure, the supporting strength is higher, and for the net-shaped supporting structure with the same size obtained by splicing, the smaller the net-shaped supporting unit which forms the net-shaped supporting structure is, the higher the supporting strength is. Therefore, after comprehensive consideration of the embodiments of the present invention, it is proposed that the area of the first net-shaped supporting unit is less than 130mm 2 。
In an embodiment of the present invention, the shape of the first mesh-shaped supporting unit is: any one of regular hexagon, circle, regular quadrangle and regular trilateral; the aperture shape of the first reticular supporting unit is as follows: any one of regular hexagon, circle, regular quadrangle and regular trilateral.
In the embodiment of the present invention, in practice, the mesh-shaped supporting unit having any shape and any aperture shape may be selected according to the production difficulty and cost, which is not particularly limited in the present invention.
In the embodiment of the present invention, in consideration that in the leukocyte filtering step, autologous blood is filtered only by gravity, and the number of leukocytes in the autologous blood is large, a densely arranged cell layer is quickly formed above the first flexible microporous array filter membrane, which prevents the filtering from continuing, the embodiment of the present invention provides: and an external force orthogonal to the flow direction of the fluid in the leukocyte removal module is intermittently applied through the vortex oscillator, and the cell layers densely arranged above the filter membrane are dispersed, so that the filter holes of the first flexible microporous array filter membrane are always kept in a smooth state, and the aim of continuously treating large-volume autologous blood can be fulfilled.
In an embodiment of the present invention, the operating conditions of the vortex oscillator include: the rotation speed is 200-3000rpm, and the vortex oscillator is periodically operated at 30s intervals.
In an embodiment of the present invention, before the leukocyte filtering step, the method may further include:
a first flexible microporous array filter membrane treatment step: and after the soaking treatment, diluting and removing the alcohol attached to the first flexible microporous array filter membrane by using a phosphate buffer solution.
By adopting the autologous blood treatment method provided by the embodiment of the invention, the white blood cells in the autologous blood can be effectively removed to obtain the filtered blood, and the filtered blood can be directly used for clinical application of autologous blood transfusion under the condition of not cleaning red blood cells.
In an embodiment of the present invention, another method for treating autoblood is provided, and fig. 2 is a flowchart of another method for treating autoblood in an embodiment of the present invention; as shown in fig. 2, the method comprises the steps of:
s201, autologous blood collection: and collecting autologous blood of the user and storing the autologous blood into the first container.
S202, transferring the autologous blood from the first container to a sample temporary storage container in the leukocyte removal module.
S203, leukocyte filtration step: under the action of external force applied by the vortex oscillator, the autologous blood is filtered by the first flexible micropore array filter membrane in the leukocyte removal module, and the filtered blood is collected and stored in the second container.
Steps S201 to S203 are similar to steps S101 to S103, and the embodiment of the present invention is not described herein again.
In an embodiment of the present invention, the red blood cells in the filtered blood may be washed, in which case the method further comprises the following steps:
s204, transferring the filtered blood from the second container to a sample temporary storage container in the red blood cell cleaning module.
In an alternative embodiment of the present invention, the leukocyte filtering module and the erythrocyte washing module may be used in conjunction, so that the temporary sample storage containers in the second container and the erythrocyte washing module are the same container, in this case, after the leukocyte filtering is completed, the filtered blood obtained by filtering directly enters the erythrocyte washing module to perform the erythrocyte washing step.
S205, a red blood cell cleaning step: and continuously adding a red blood cell cleaning solution into the red blood cell cleaning module under the action of external force applied by the vortex oscillator, and cleaning the filtered blood by utilizing a second flexible micropore array filter membrane in the red blood cell cleaning module.
Wherein, external force orthogonal in the fluidic flow direction in the red blood cell cleaning module, the flexible micropore array filter membrane of second is used for catching red blood cells, the filtration pore aperture of the flexible micropore array filter membrane of second is 2-4 microns, and the filtration pore interval is 2 microns, and thickness is not more than 10 microns, the netted bearing structure of second has been laid to the flexible micropore array filter membrane of second, the netted bearing structure of second is used for supporting the flexible micropore array filter membrane of second, the netted bearing structure of second comprises the netted supporting element of a plurality of seconds, the netted supporting element evenly distributed of a plurality of seconds is in the flexible micropore array filter membrane below of second, the aperture of the netted supporting element of second is greater than the filtration pore aperture of the second micropore array filter membrane, the area of the netted supporting element of second is less than 25mm 2 。
In the embodiment of the invention, the erythrocyte cleaning module is used for cleaning erythrocytes in filtered blood treated by the leukocyte removing module, and reducing or even removing pollutants such as plasma, free hemoglobin and activated blood coagulation factors. The working principle is that a flexible microporous array filter membrane with the pore diameter smaller than the size of red blood cells is selected, cleaning solution is added for multiple times and is filtered, components with the size smaller than the size of the red blood cells in blood after filtration are removed in a mode of replacing filtrate, and the red blood cells are cleaned.
In an embodiment of the present invention, the cleaning solution is physiological saline, and in a specific implementation, the volume of the cleaning solution may be determined according to a requirement of red blood cell cleaning, and in a preferred embodiment of the present invention, the volume of the cleaning solution is 1/2 of the volume of the autologous blood. In the embodiment of the invention, the volume of the cleaning solution influences the cleaning degree of the red blood cells, the larger the volume of the cleaning solution is, the better the cleaning effect is, but the longer the treatment time is, so that in the embodiment of the invention, the volume of the cleaning solution is determined to be 1/2 of the volume of the autologous blood under comprehensive consideration.
In the embodiment of the invention, the red blood cells are round cake-shaped cells with the diameter of about 6 microns and the thickness of about 2 microns, theoretically, when the size of the filter hole of the filter membrane is 4 microns, the red blood cells cannot pass through the filter membrane, the filtrate can smoothly pass through the filter membrane, and the smaller the size of the filter hole is, the more difficult the red blood cell loss is. However, in the embodiment of the present invention, after completing the red blood cell washing, an external force orthogonal to the plane of the second micropore array filter is applied to deform the red blood cells through the filter, so as to complete the red blood cell recovery. Therefore, the size of the filtration pores of the second microporous array filtration membrane is determined to be 2 to 4 μm in combination.
In the embodiment of the invention, similarly to the determination method of the first flexible micropore array filter membrane, the filtration effectiveness of the filter pores and the overall strength of the filter membrane are also considered, and under the condition that the size of the filter pores of the second flexible micropore array filter membrane is 2-4 micrometers, the filter pore spacing is determined to be 2 micrometers. Also in order to reduce the cell travel of the red blood cells during filtration to reduce red blood cell damage, the second flexible micro-pore array filter membrane is provided with a thickness of no more than 10 microns.
In the embodiment of the invention, the area of the second flexible micropore array filter membrane is the same as that of the first flexible micropore array filter membrane, so that the blood after filtration can be conveniently treated.
In embodiments of the invention, a mesh support structure is also laid underneath the second flexible micro-well array filter membrane, based on similar considerations as the first flexible micro-well array filter membrane. The difference is that in the embodiment of the present invention, since an external force orthogonal to the plane of the second microporous array filter membrane is to be applied subsequently to deform the red blood cells passing through the filter membrane to complete the recovery of the red blood cells, the requirement on the strength of the second flexible microporous array filter membrane is higher, and accordingly, the requirement on the supporting function of the mesh-shaped supporting structure is higher. Therefore, the embodiment of the present invention provides that the area of the second net-shaped supporting unit is less than 25mm 2 。
In an embodiment of the present invention, the shape of the second mesh-like supporting unit may also be: any one of regular hexagon, circle, regular quadrangle and regular trilateral; the aperture shape of the second reticular supporting unit can also be as follows: any one of regular hexagon, circle, regular quadrangle and regular trilateral.
In the embodiment of the present invention, in practice, the mesh-shaped supporting unit having any shape and any aperture shape may be selected according to the production difficulty and cost, which is not particularly limited in the present invention.
In the embodiment of the invention, based on similar consideration in the white blood cell filtering step, the external force which is applied to the flow direction of the fluid in the red blood cell cleaning module by the vortex oscillator is also set to be applied intermittently, and the densely arranged cell layer formed above the filter membrane is dispersed, so that the aim of continuously treating the large-volume autologous blood is fulfilled.
In an embodiment of the present invention, the operating conditions of the vortex oscillator include: the rotation speed is 200-3000rpm, and the vortex oscillator is periodically operated at 30s intervals.
In an embodiment of the present invention, before the red blood cell washing step, the method further comprises:
a second flexible microporous array filter membrane treatment step: and performing infiltration treatment on the second flexible microporous array filter membrane by using alcohol, and after the infiltration treatment, diluting and removing the alcohol attached to the second flexible microporous array filter membrane by using a phosphate buffer solution.
S206, the liquid outlet side of the red blood cell cleaning module is connected with a negative pressure module, and a red blood cell collecting module is arranged and used for storing the processed red blood cells.
In the embodiment of the invention, after the red blood cells are cleaned, the negative pressure module and the red blood cell collecting module can be used for collecting the red blood cells so as to perform subsequent clinical application.
S207, a red blood cell collecting step: and applying an external force orthogonal to the plane of the second microporous array filter membrane to the red blood cell cleaning module by using the negative pressure module, so that the red blood cells pass through the filter pores of the second flexible microporous array filter membrane and reach the red blood cell collecting module, and the red blood cells captured on the second flexible microporous array filter membrane are collected to the red blood cell collecting module.
In the embodiment of the invention, an external force orthogonal to the plane of the second micropore array filter membrane can be applied to deform the red blood cells to pass through the filter membrane so as to complete the recovery of the red blood cells. In the process, because the red blood cells have small diameters and are easy to deform and can easily pass through the filter membrane, the white blood cells have large diameters and relatively weak deformability, and the white blood cells which are not completely filtered in the white blood cell filtering step can be further removed by collecting the red blood cells through the filter membrane.
In an optional embodiment, the negative pressure module is a syringe pump, and S206 specifically includes: the liquid outlet side of the red blood cell cleaning module is connected with an injection pump, the injection pump comprises an injector, the injector is used for temporarily storing red blood cells, and the liquid outlet of the injector is connected with the red blood cell collecting module.
S207 specifically includes: under the action of an external force applied by the injection pump and orthogonal to the plane of the second microporous array filter membrane, the red blood cells pass through the filter pores of the second flexible microporous array filter membrane and are temporarily stored in an injector connected with the injection pump, and then the red blood cells are transferred from the injector to the red blood cell collection module, so that the red blood cells captured on the second flexible microporous array filter membrane are collected in the red blood cell collection module.
In the embodiment of the invention, in order to further facilitate subsequent use, the negative pressure module can be set as an injection pump, and the red blood cell collection module is set as a blood bag, so that the red blood cells cleaned on the second flexible microporous array filter membrane can be directly transferred into the blood bag through the injection pump for storage.
In an optional embodiment of the present invention, the autologous blood treatment method further comprises the steps of:
s208, evaluation step of autoblood treatment effect: and taking out the second microporous array filter membrane from the erythrocyte washing module, dyeing and counting leucocytes of the second flexible microporous array filter membrane, counting the number of leucocytes captured by the second flexible microporous array filter membrane, and judging whether the number of leucocytes is smaller than a preset value or not to obtain an evaluation result.
In the embodiment of the invention, after the collection of the red blood cells is completed, the second flexible microporous array filter membrane can be taken out from the red blood cell cleaning module, the second flexible microporous array filter membrane is dyed, the dyed second flexible microporous array filter membrane is placed under a microscope for counting white blood cells, and the number of the white blood cells captured by the second flexible microporous array filter membrane is counted to estimate the removal effect of the white blood cells in the autologous blood treatment process. Theoretically, the autologous blood treatment effect can also be evaluated by performing staining counting on the first flexible micropore array filter membrane in the leukocyte removal module, but most of leukocytes and a few of red blood cells in the autologous blood are captured on the first flexible micropore array filter membrane, the number of the cells is too large, the cells may be distributed in a multi-layer 3D space, the observation and counting difficulty is high, and therefore in order to reduce the evaluation difficulty, in the embodiment of the invention, the autologous blood treatment effect is evaluated by using the second flexible micropore array filter membrane in the red blood cell cleaning module.
In the embodiment of the present invention, the staining step may be performed by using a standard method, in this embodiment, a fluorescence dye of Hoechst + Cell Tracker is used to mark the Cell nucleus and the Cell cytoplasm, respectively, and a lius dye, a reishi dye, and the like may also be used, which is not specifically limited in the present invention.
Prior to the red blood cell washing step, the method further comprises:
a second flexible microporous array filter membrane treatment step: and performing infiltration treatment on the second flexible microporous array filter membrane by using alcohol to improve the hydrophilicity, and after the infiltration treatment, diluting and removing the alcohol attached to the second flexible microporous array filter membrane by using a phosphate buffer solution.
In a second aspect, embodiments of the present invention further provide an autologous blood treatment apparatus, including: the device comprises a first container, a leukocyte removal module, a second container, a red blood cell cleaning module, a vortex oscillator, a negative pressure module and a red blood cell collecting module; the device is used for executing the autologous blood treatment method of the first aspect.
In an embodiment of the present invention, a first mesh-shaped supporting structure is disposed in the leukocyte removal module, the first mesh-shaped supporting structure is used for supporting the first flexible microporous array filter membrane, the first mesh-shaped supporting structure is composed of a plurality of first mesh-shaped supporting units, the plurality of first mesh-shaped supporting units are uniformly distributed below the first flexible microporous array filter membrane, the pore size of each first mesh-shaped supporting unit is larger than the pore size of a pore of the first flexible microporous array filter membrane, and the area of each first mesh-shaped supporting unit is smaller than 130mm 2 ；
The second netted bearing structure is used for supporting the flexible micropore array filter membrane of second, the netted bearing structure of second comprises the netted supporting element of a plurality of second, a plurality of netted supporting element evenly distributed of second are in the flexible micropore array filter membrane below of second, the aperture of the netted supporting element of second is greater than the filtration pore aperture of the flexible micropore array filter membrane of second, the area of the netted supporting element of second is less than 25mm 2 。
In practical applications, the autologous blood treatment device may be configured as an integrated device, as shown in fig. 3, and fig. 3 is a schematic view of an autologous blood treatment device provided according to an embodiment of the present invention. The device mainly comprises four parts, namely a leukocyte removal unit, an erythrocyte cleaning unit, an erythrocyte recovery unit and an auxiliary structure. Specifically, the leukocyte removal unit is composed of a sample temporary storage 11, a first flexible microporous filter membrane and fixed carrier 12, a vortex oscillator 13, an electromagnetic valve 14 and a sample outlet 15. The erythrocyte cleaning unit consists of an erythrocyte cleaning solution inlet 21, an electromagnetic valve 22, a sample temporary storage 23, a second flexible microporous filter membrane, a fixed carrier 24 and a vortex oscillator 25. The red blood cell recovery unit consists of an electromagnetic valve 31, a vacuum pump interface and a sample outlet 32. The auxiliary structure comprises a pipeline 41 consisting of a silica gel hose and an instrument shell 42.
In an embodiment of the present invention, the sample outlet 32 of the red blood cell recovery unit of the autologous blood processing apparatus may be further connected to a syringe pump through a silicone hose to return the filtered and recovered red blood cells to the user.
In the embodiment of the invention, the autologous blood treatment device is an integrated device, the volume of the device is matched with that of the filter membrane, the device is small in volume and convenient to carry, and the device can realize efficient filtration and recovery of autologous blood based on the leukocyte removal unit, the erythrocyte cleaning unit and the erythrocyte recovery unit.
Therefore, in the embodiment of the invention, the integrated portable autologous blood treatment device can be used for efficiently collecting, filtering and returning the autologous blood in a severe environment (such as a battlefield, a severe environment far away from a medical institution and the like).
In order to make the present invention more understandable to those skilled in the art, the method for treating autologous blood provided by the present invention is described below by way of specific examples.
Example 1
The aperture is 6um, the distance between the holes is 2um, the thickness is 10um, and the effective filtration area is about 130mm 2 The leukocyte-removing filtration operation was performed on 2mL of autologous blood by applying an additional vortex at a rotation speed of 2000r for a duration and at intervals of 30s. Then the aperture is 4um, the distance between the holes is 2um, the thickness is 10um, and the effective filtration area is about 130mm 2 Second flexible micro-holeThe filter membrane was arrayed, and negative pressure was applied to the filter membrane with a 2mL blood collection tube, and the filtrate obtained after the leukocyte filtration operation was subjected to an erythrocyte washing and collection operation. The whole treatment process is completed within 5 min.
After completion of the treatment, the results of staining the first and second flexible microwell array filters with Hoechst and Cell Tracker stains are shown in fig. 4 (a) and (b). After staining, hoechst binds to the nucleus and fluoresces blue, while Cell Tracker binds to the cytoplasm and fluoresces red. Therefore, the processed white blood cells emitting blue and red fluorescence simultaneously are intact white blood cells, the processed white blood cells emitting only blue fluorescence are cell nucleus fragments, and the processed white blood cells emitting only red fluorescence are cytoplasm fragments or red blood cells.
As can be seen from the figure, most of the white blood cells can be captured on the first flexible micropore array filter membrane, and the first flexible micropore array filter membrane is stacked along the thickness direction of the filter membrane, so that the number cannot be counted; and the second flexible micropore array filter membrane only captures a small amount of complete leucocytes and cell nucleus fragments, which proves that the first flexible micropore array filter membrane has a better leucocyte filtration effect.
Example 2
Table 1 is a comparison of results of autologous blood (whole blood) samples treated using the autologous blood treatment method provided by the present invention and the commonly used Controlled Incremental Filtration (CIF) method, inertial flow method, and Deterministic Lateral Displacement (DLD) method, respectively.
TABLE 1 comparison of treatment results of autologous blood (whole blood) treatment methods
As can be seen from Table 1, the late Controlled Incremental Filtration (CIF) method, the inertial flow and Deterministic Lateral Displacement (DLD) method, leukocyte depletion and erythrocyte transfusionThe flux is high, but the three methods can only process blood after pretreatment such as dilution, centrifugation and the like, and the universality is limited; while the early Controlled Incremental Filtration (CIF) method can directly process whole blood, it has the disadvantage of lower red blood cell output flux than the autologous blood processing method provided by the embodiments of the present invention. In addition, in the embodiment of the invention, the filtration area of the first flexible micropore array filter membrane and the filtration area of the second flexible micropore array filter membrane are 130mm 2 In this case, it takes about 7.5 minutes to treat 2mL of autologous blood, and if the area of the flexible microporous array filter is expanded by about 50 times to 8000mm 2 Accordingly, the filtering of the corresponding 100mL autologous blood sample can be completed in about 7.5 minutes, and accordingly, the processing speed of the autologous blood processing method provided by the present embodiment can reach 13mL/min, which is significantly higher than that of other methods.
Therefore, the autologous blood treatment method provided by the embodiment of the invention can be used for rapidly treating the autologous blood sample with large volume, does not need to pretreat the autologous blood, and can achieve the treatment effects of high leukocyte removal rate and high erythrocyte activity.
For simplicity of explanation, the method embodiments are described as a series of acts or combinations, but those skilled in the art will appreciate that the present invention is not limited by the order of acts, as some steps may occur in other orders or concurrently in accordance with the invention. Further, those skilled in the art will appreciate that the embodiments described in the specification are preferred embodiments and that the acts and elements referred to are not necessarily required to practice the invention.
The method and the device for processing the autologous blood provided by the invention are described in detail, and the principle and the implementation mode of the invention are explained by applying specific examples, and the description of the examples is only used for helping to understand the method and the core idea of the invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present invention.
Claims (10)
1. A method of autologous blood treatment, said method comprising:
and (3) autologous blood collection step: collecting blood of a user and storing the blood into a first container;
transferring the autologous blood from the first container to a sample temporary storage container in a leukocyte removal module;
a leukocyte filtration step: under the effect of external force that vortex oscillator was applyed, first flexible micropore array filter membrane in the module is got rid of to leucocyte is right autoblood filters, collects and filters back blood, deposits to the second container, wherein, external force quadrature in the fluidic flow direction in the module is got rid of to leucocyte, first flexible micropore array filter membrane is used for the filtering leucocyte in the autoblood, the filtration pore diameter of first flexible micropore array filter membrane is 6-10 microns, and the filtration pore spacing is 2-4 microns, and thickness is not more than 10 microns, first netted bearing structure has been laid under the first flexible micropore array filter membrane, first netted bearing structure is used for supporting first flexible micropore array filter membrane, first netted bearing structure comprises a plurality of first netted supporting unit, a plurality of first netted supporting unit evenly distributed in first flexible micropore array filter membrane below, the aperture of first netted supporting unit is greater than the filtration pore diameter of first flexible micropore array filter membrane, the area of first supporting unit is less than 130 mm's filter membrane, the area of first supporting unit is less than 130mm 2 。
2. The autologous blood treatment method of claim 1, further comprising:
transferring the filtered blood from the second container to a sample buffer container in a red blood cell washing module;
and (3) red blood cell cleaning: continuously adding a red blood cell cleaning solution into the red blood cell cleaning module under the action of an external force applied by the vortex oscillator, and cleaning the filtered blood by using a second flexible micropore array filter membrane in the red blood cell cleaning module, wherein the external force is orthogonal to the flow direction of fluid in the red blood cell cleaning module, and the second flexible micropore array filter membrane is flexible and flexibleThe hole array filter membrane is used for catching red blood cells, the filtration pore aperture of the flexible micropore array filter membrane of second is 2-4 microns, and the filtration pore interval is 2 microns, and thickness is not more than 10 microns, the netted bearing structure of second has been laid under the flexible micropore array filter membrane of second, the netted bearing structure of second is used for supporting the flexible micropore array filter membrane of second, the netted bearing structure of second comprises a plurality of second netted supporting units, the netted supporting unit evenly distributed of a plurality of second is in the flexible micropore array filter membrane below of second, the aperture of the netted supporting unit of second is greater than the filtration pore aperture of the flexible micropore array filter membrane of second, the area of the netted supporting unit of second is less than 25mm 2 ；
The liquid outlet side of the red blood cell cleaning module is connected with a negative pressure module, and is provided with a red blood cell collecting module which is used for storing the treated red blood cells;
and (3) red blood cell collection: and applying an external force orthogonal to the plane of the second flexible microporous array filter membrane to the red blood cell cleaning module by using the negative pressure module, so that the red blood cells pass through the filter pores of the second flexible microporous array filter membrane and reach the red blood cell collecting module, and the red blood cells captured on the second flexible microporous array filter membrane are collected to the red blood cell collecting module.
3. The autologous blood treatment method of claim 2, wherein the negative pressure module is a syringe pump, the outlet side of the red blood cell washing module is connected to the negative pressure module, and a red blood cell collecting module is provided, comprising:
the liquid outlet side of the red blood cell cleaning module is connected with an injection pump, the injection pump comprises an injector, the injector is used for temporarily storing red blood cells, and the liquid outlet of the injector is connected with the red blood cell collecting module;
the red blood cell collecting step comprises: under the action of an external force applied by the injection pump and orthogonal to the plane of the second flexible microporous array filter membrane, the red blood cells pass through the filter pores of the second flexible microporous array filter membrane and are temporarily stored in an injector connected with the injection pump, and then the red blood cells are transferred from the injector to the red blood cell collection module, so that the red blood cells captured on the second flexible microporous array filter membrane are collected to the red blood cell collection module.
4. The autologous blood treatment method according to claim 1 or 2, wherein the shape of the first mesh-like support unit or the shape of the second mesh-like support unit is: any one of regular hexagon, circle, regular quadrangle and regular trilateral;
the aperture shape of the first net-shaped supporting unit or the aperture shape of the second net-shaped supporting unit is as follows: any one of regular hexagon, circle, regular quadrangle and regular trilateral.
5. The autologous blood treatment method according to claim 2, wherein after the red blood cell collection step, the method further comprises:
evaluation step of autoblood treatment effect: and taking out the second flexible micropore array filter membrane from the erythrocyte cleaning module, dyeing and counting leukocytes of the second flexible micropore array filter membrane, counting the number of the leukocytes captured by the second flexible micropore array filter membrane, judging whether the number of the leukocytes is smaller than a preset value or not, and obtaining an evaluation result.
6. The autologous blood treatment method according to claim 2, wherein before the leukocyte filtration step, the method further comprises: a first flexible microporous array filter membrane treatment step: performing infiltration treatment on the first flexible microporous array filter membrane by using alcohol, and after the infiltration treatment, diluting and removing the alcohol attached to the first flexible microporous array filter membrane by using a phosphate buffer solution;
prior to the red blood cell washing step, the method further comprises:
a second flexible microporous array filter membrane treatment step: and performing infiltration treatment on the second flexible microporous array filter membrane by using alcohol, and after the infiltration treatment, diluting and removing the alcohol attached to the second flexible microporous array filter membrane by using a phosphate buffer solution.
7. The autologous blood treatment method of claim 1, wherein the operating conditions of the vortex oscillator include: the rotation speed is 200-3000rpm, and the vortex oscillator is periodically operated at 30s intervals.
8. The method according to claim 2, wherein in the red blood cell washing step, the washing solution is physiological saline, and the volume of the washing solution is 1/2 of the volume of the autologous blood.
9. An autologous blood treatment apparatus, characterized in that said apparatus comprises:
the device comprises a first container, a white blood cell removing module, a second container, a red blood cell cleaning module, a vortex oscillator, a negative pressure module and a red blood cell collecting module;
the device is used for executing the autologous blood treatment method of any one of claims 1-8.
10. The autologous blood treatment apparatus of claim 9, wherein the leukocyte removal module is provided with a first mesh-like support structure, the first mesh-like support structure is used for supporting the first flexible microporous array filter membrane, the first mesh-like support structure is composed of a plurality of first mesh-like support units, the plurality of first mesh-like support units are uniformly distributed below the first flexible microporous array filter membrane, the pore size of the first mesh-like support units is larger than that of the first flexible microporous array filter membrane, and the area of the first mesh-like support units is smaller than 130mm 2 ；
The second net-shaped supporting structure is used for supporting the second flexible micropore array filter membrane, the second net-shaped supporting structure consists of a plurality of second net-shaped supporting units, the plurality of second net-shaped supporting units are uniformly distributed below the second flexible micropore array filter membrane, and the second net-shaped supporting unitThe aperture of the first net-shaped supporting unit is larger than the aperture of the filtration pores of the second flexible microporous array filtration membrane, and the area of the second net-shaped supporting unit is smaller than 25mm 2 。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210832461.XA CN115141800B (en) | 2022-07-15 | 2022-07-15 | Autologous blood treatment method and device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210832461.XA CN115141800B (en) | 2022-07-15 | 2022-07-15 | Autologous blood treatment method and device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115141800A true CN115141800A (en) | 2022-10-04 |
| CN115141800B CN115141800B (en) | 2024-04-16 |
Family
ID=83412761
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210832461.XA Active CN115141800B (en) | 2022-07-15 | 2022-07-15 | Autologous blood treatment method and device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115141800B (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020058030A1 (en) * | 2000-05-03 | 2002-05-16 | Eligix, Inc. | Whole blood separator apparatus and method of use |
| CN102430163A (en) * | 2011-09-02 | 2012-05-02 | 四川大学华西医院 | Method for recovering intraoperative autoblood |
| CN105999445A (en) * | 2016-08-02 | 2016-10-12 | 宁波市妇女儿童医院 | Method for treating recycled blood in cesarean section |
| CN107674861A (en) * | 2017-11-02 | 2018-02-09 | 苏州浚惠生物科技有限公司 | Method for separating and detecting single cell level of rare cell |
| CN112292161A (en) * | 2018-06-20 | 2021-01-29 | 意大利费森尤斯血液技术有限公司 | Filter assembly and container for collecting blood comprising a filter assembly |
| CN113252428A (en) * | 2021-04-28 | 2021-08-13 | 北京大学第三医院（北京大学第三临床医学院） | Filter device and liquid sample processing system |
| US20210299272A1 (en) * | 2020-03-31 | 2021-09-30 | The Trustees Of The University Of Pennsylvania | Methods and compositions for drug delivery |
-
2022
- 2022-07-15 CN CN202210832461.XA patent/CN115141800B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020058030A1 (en) * | 2000-05-03 | 2002-05-16 | Eligix, Inc. | Whole blood separator apparatus and method of use |
| CN102430163A (en) * | 2011-09-02 | 2012-05-02 | 四川大学华西医院 | Method for recovering intraoperative autoblood |
| CN105999445A (en) * | 2016-08-02 | 2016-10-12 | 宁波市妇女儿童医院 | Method for treating recycled blood in cesarean section |
| CN107674861A (en) * | 2017-11-02 | 2018-02-09 | 苏州浚惠生物科技有限公司 | Method for separating and detecting single cell level of rare cell |
| CN112292161A (en) * | 2018-06-20 | 2021-01-29 | 意大利费森尤斯血液技术有限公司 | Filter assembly and container for collecting blood comprising a filter assembly |
| US20210299272A1 (en) * | 2020-03-31 | 2021-09-30 | The Trustees Of The University Of Pennsylvania | Methods and compositions for drug delivery |
| CN113252428A (en) * | 2021-04-28 | 2021-08-13 | 北京大学第三医院（北京大学第三临床医学院） | Filter device and liquid sample processing system |
Non-Patent Citations (5)
| Title |
|---|
| TSAU-MAU CHOU: "Autologous platelet concentrates in maxillofacial regenerative therapy", KAOHSIUNG J MED SCI, vol. 36, no. 5 * |
| 曹甜甜: "回收式自体血在产科的应用现状及研究进展", 实用妇产科杂志, vol. 36, no. 11 * |
| 李志强: "简明临床输血理论与实践", 28 February 2010, 上海世界图书出版公司, pages: 367 * |
| 李慧文: "献血与志愿服务", 31 August 2011, 科学普及出版社, pages: 144 * |
| 袁红: "临床用血管理制度及技术操作规程范文"", 31 March 2022, 重庆大学出版社, pages: 9 - 6 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115141800B (en) | 2024-04-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5249973B2 (en) | Ascites treatment system and cleaning method thereof | |
| JPH03173825A (en) | Filter for purifying blood platelet | |
| JP6707031B2 (en) | Cell separation material and cell separation method | |
| JPWO2002101029A1 (en) | Method for separating and enriching cells for kidney regeneration | |
| JP2001500753A (en) | Method for removing tumor cells from stem cell products contaminated with tumor cells | |
| JP4107410B2 (en) | Monocyte capture filter | |
| JP5336109B2 (en) | Methods for enriching mononuclear cells and platelets | |
| JP2001061955A (en) | Autotransfusion device and method | |
| JP6645605B2 (en) | Filtration device and filtration method | |
| JP5785713B2 (en) | Leukocyte fraction collection device | |
| KR20010075549A (en) | Biological fluid filter and system | |
| JP2013036818A (en) | Method of concentrating tumor cell and separation material | |
| CN115141800B (en) | Autologous blood treatment method and device | |
| JP2010088376A (en) | Cell recovering device and liquid containing adipose-derived cell | |
| JP2002087971A (en) | Method for separating living body tissue-regenerating cell and device for the same | |
| JP2003304865A (en) | Method for separating cell | |
| CN109012207B (en) | Cross flow filtration system for enriching circulating tumor cells | |
| JPH03284263A (en) | Device for collecting plasma and concentrated erythrocyte | |
| JP2012210187A (en) | Method for condensing cell suspension | |
| JP2012080821A (en) | Method of concentrating nucleated cell component in blood | |
| JP4162128B2 (en) | Sample for cytodiagnosis, preparation method and apparatus thereof | |
| JP4362529B2 (en) | Monocyte separation culture apparatus and dendritic cell recovery method | |
| JPH11290060A (en) | Cell separation filter suitable for recovering cell, cell separation system and separation of cell | |
| CN201049099Y (en) | Filtering film in blood therapeutic instrument | |
| JP3945725B2 (en) | Cell separation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |