CN114410580A - Extraction method of PBMC (peripheral blood mononuclear cell) - Google Patents
Extraction method of PBMC (peripheral blood mononuclear cell) Download PDFInfo
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- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 title claims abstract description 28
- 238000000605 extraction Methods 0.000 title claims abstract description 23
- 239000000243 solution Substances 0.000 claims abstract description 35
- 210000004027 cell Anatomy 0.000 claims abstract description 29
- 210000004369 blood Anatomy 0.000 claims abstract description 22
- 239000008280 blood Substances 0.000 claims abstract description 22
- 238000000926 separation method Methods 0.000 claims abstract description 19
- 239000012981 Hank's balanced salt solution Substances 0.000 claims abstract description 11
- 210000004698 lymphocyte Anatomy 0.000 claims abstract description 11
- 238000004140 cleaning Methods 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 8
- 239000006285 cell suspension Substances 0.000 claims abstract description 6
- 238000001514 detection method Methods 0.000 claims abstract description 5
- 230000000694 effects Effects 0.000 claims abstract description 5
- 239000011259 mixed solution Substances 0.000 claims abstract description 5
- 238000007865 diluting Methods 0.000 claims abstract description 3
- 238000005119 centrifugation Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 239000012997 ficoll-paque Substances 0.000 claims description 8
- 230000001133 acceleration Effects 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 5
- 210000003743 erythrocyte Anatomy 0.000 claims description 4
- 210000000265 leukocyte Anatomy 0.000 claims description 4
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 210000005087 mononuclear cell Anatomy 0.000 claims description 2
- 230000003833 cell viability Effects 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 210000001616 monocyte Anatomy 0.000 description 3
- 230000035899 viability Effects 0.000 description 3
- 231100000747 viability assay Toxicity 0.000 description 3
- 238000003026 viability measurement method Methods 0.000 description 3
- 230000006037 cell lysis Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000000405 serological effect Effects 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000002619 cancer immunotherapy Methods 0.000 description 1
- 230000006727 cell loss Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000000432 density-gradient centrifugation Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000002825 functional assay Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
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- 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
- C12N5/0645—Macrophages, e.g. Kuepfer cells in the liver; Monocytes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N5/0635—B lymphocytes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N5/0636—T lymphocytes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N5/0646—Natural killers cells [NK], NKT cells
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
- C12N2509/10—Mechanical dissociation
Abstract
The invention relates to the technical field of PBMC detection, in particular to a PBMC extraction method, which comprises the following steps of S1: adding a sample diluted by whole blood into a centrifuge tube, adding an HBSS solution, and performing 1: 1, diluting; s2: slowly adding the diluted blood sample into a centrifuge tube filled with lymphocyte separation liquid to ensure that the blood sample is on the upper layer of the lymphocyte separation liquid; s3: placing the solution obtained in the step S2 at room temperature, and performing horizontal centrifugal separation to separate the solution into layers; s4: taking a white film layer, adding a cleaning solution, and cleaning; s5: performing centrifugal separation on the mixed solution obtained in the step S4, and re-suspending cells to obtain a cell suspension; s6: taking the cell suspension, and carrying out cell counting and activity detection. The whole blood sample is divided into a plurality of parts, and the parts are respectively extracted and then combined, so that the influence of operation on the extraction result is reduced. Compared with the prior art, the extraction method has higher cell yield and repeatability by controlling the experimental conditions in the extraction method.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to the technical field of PBMC detection, in particular to a PBMC extraction method.
[ background of the invention ]
Peripheral Blood Mononuclear Cells (PBMC) include lymphocytes and monocytes, which in turn include T cells, B cells and Natural Killer (NK) cells. PBMCs are the most commonly used cell models in immunological functional studies, such as cell proliferation, cytotoxicity, cytokine secretion, etc. For example, in the field of cancer immunotherapy, it is desirable to isolate PBMC from whole patient blood, and the isolated PBMC is further amplified or subjected to different functional assays.
At present, density gradient centrifugation is mostly adopted for extraction of PBMC. However, in clinical trials, the total number of cells obtained varies greatly depending on experimental conditions such as separation fluid, centrifugal force, centrifugation time, and the like.
[ summary of the invention ]
The invention aims to provide a PBMC extraction method, which is suitable for a PBMC extraction process of whole blood and overcomes the problem of low cell yield.
In order to achieve the purpose, the invention adopts the following technical scheme:
a PBMC extraction method is characterized in that: comprises the following steps of (a) carrying out,
s1: adding a whole blood sample into a centrifuge tube, adding an HBSS solution, and performing 1: 1, diluting;
s2: slowly adding the diluted blood sample into a centrifuge tube filled with lymphocyte separation liquid to ensure that the blood sample is on the upper layer of the lymphocyte separation liquid;
s3: placing the solution obtained in the step S2 at room temperature, performing horizontal centrifugal separation to layer the solution, and sequentially including a red blood cell layer, a separation liquid layer, a leukocyte layer (mononuclear cell layer), and a diluted plasma layer from bottom to top;
s4: taking a white film layer, adding a cleaning solution, and cleaning;
s5: centrifuging the mixed solution obtained in the step S4, removing supernatant, adding a cleaning solution, and resuspending cells to obtain a cell suspension;
s6: taking the cell suspension, and carrying out cell counting and activity detection.
As a further improvement of the present invention, in step S1, the sample after dilution of whole blood is divided into at least 2 parts, and extraction processes are respectively performed, and finally obtained extraction solutions are combined.
As a further improvement of the invention, the lymphocyte separation solution is a Ficoll Paque solution.
As a further improvement of the invention, the conditions of the horizontal centrifugation of the step S3 are that the braking mode is cancelled, the rotating speed is 790g, and the centrifugation is carried out for 20 minutes.
As a further improvement of the present invention, the cleaning solution in step S4 is HBSS solution.
As a further improvement of the invention, the conditions of the centrifugation in the step S5 are that the maximum acceleration and braking mode is set, the room temperature is 290g, and the centrifugation is carried out for 10 minutes.
Compared with the prior art, the invention has the beneficial effects that: the whole blood sample is divided into at least 2 parts, and after extraction treatment is respectively carried out, the whole blood sample is combined after being lifted, so that the influence on the extraction result caused by operation is reduced. The control of the centrifugation conditions in step S2 makes the mixed solution easier to separate into layers between solutions of different densities, and avoids excessive loss of lymphocytes. The control of the centrifugal force in step S5 avoids the risk of cell lysis. Compared with the existing method, the extraction method of the invention has higher cell yield and repeatability.
[ appendix ] description
FIG. 1 is a graph comparing the total number of PBMC cells of examples of the present invention with those of comparative examples.
FIG. 2 is a graph comparing PBMC viability of examples of the present invention and comparative examples.
[ detailed description ] embodiments
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The reagents used in the following examples are as follows:
| reagent | Manufacturer of the product | Goods number | Storage conditions |
| Ficoll Paque | GE Healthcare | 17-5442-02 | 2-8℃ |
| HBSS | gibco | 14170-112 | 2-8℃ |
| AO/PI dye liquor | CountStar | RE010212 | 2°C-8°C |
Example 1
A PBMC extraction method specifically comprises the following steps:
a) 3 15mL centrifuge tubes were prepared for each sample in advance and labeled with sample information.
b) In a biosafety cabinet, 5mL of ficoll Paque solution was added to 2 15mL centrifuge tubes for each sample using a serum pipette for use.
c) The diluted blood sample was slowly added by electronic pipette along the tube wall into the 2 15mL centrifuge tubes containing the Ficoll Paque solution, each tube was filled to 15mL and the blood sample was kept on the top layer of the separation medium, forming a clear layered interface.
d) The centrifuge tube was placed in a centrifuge horizontal rotor and the braking mode was removed and centrifuged at room temperature 790g for 20 minutes.
e) After centrifugation, the layering effect was observed. The red blood cells should sink to the bottom layer, which is followed by the separation layer, the leukocyte layer (monocyte layer) and the diluted plasma layer.
f) The upper diluted plasma layer is firstly sucked up and discarded until the distance between the upper diluted plasma layer and the white membrane layer is 2-3 mm.
g) The tunica albuginea cells were aspirated slowly with sterile tubing, taking care not to aspirate other layers, the aspirated tunica albuginea was transferred to a new 15mL centrifuge tube, and finally, 1 × HBSS was added to 15mL and mixed well.
h) Set the maximum acceleration and braking mode, centrifuge for 10 minutes at room temperature of 290 g.
i) The supernatant was discarded, the bottom cell pellet was flicked, and then 1 × HBSS was added again to 15mL and mixed by gentle inversion to resuspend the cells.
j) Set the maximum acceleration and braking mode, centrifuge for 10 minutes at room temperature of 290 g.
k) After discarding the supernatant, 1 × HBSS 4mL was added, and the mixture was gently flicked to the bottom of the tube to resuspend the cells.
l) the resuspended cell solutions were pooled, examined for cell count and viability and recorded in the table.
Example 2
Comparative example
a) 2 50mL and 2 15mL centrifuge tubes were prepared for each sample in advance and labeled with sample information.
b) 15mL of Ficoll Paque solution was added to 1 50mL centrifuge tube for each sample.
c) The tubes were sequentially washed with 1 × HBSS pipetted with an electronic pipetter and serological pipette (sterile) and transferred further into the corresponding 50mL centrifuge tubes described above. So that a total volume of 30mL, which is twice the volume of the Ficoll Paque solution, is finally obtained.
d) The diluted blood sample was slowly added along the tube wall by an electronic pipette and serological pipette (sterile) to the 50mL centrifuge tube containing the Ficoll Paque solution to ensure that the blood sample was completely on the upper layer of the separation solution and a clear layered interface was formed.
e) The centrifuge tube was placed in a horizontal rotor of a centrifuge, set to maximum acceleration and cancel braking mode, and centrifuged at 490g for 40 minutes at room temperature.
f) After centrifugation, the layering effect was observed. The red blood cells should sink to the bottom layer, which is followed by the separation layer, the leukocyte layer (monocyte layer) and the diluted plasma layer.
g) The upper diluted plasma layer is firstly sucked up and discarded until the distance between the upper diluted plasma layer and the white membrane layer is 2-3 mm.
h) The buffy coat cells were slowly aspirated with a sterile pipette, taking care not to aspirate other layers, and the aspirated buffy coat was transferred to a new 15mL centrifuge tube.
i) Set the maximum acceleration and braking mode, centrifuge for 10 minutes at 280g room temperature.
j) The supernatant was aspirated and transferred to a new 15mL centrifuge tube to avoid cell loss as much as possible.
k) Add 1X HBSS 1mL and flick the tube bottom to mix well to resuspend the cells.
l) cell counts and viability assays were performed and recorded in the table.
Example 3
Cell counting and viability assays
a) One EP tube was taken for each sample and the sample information was labeled.
b) Add 12. mu.L of AO/PI stain and an equal volume of sample to an EP tube and mix well.
c) The cell counting plate was removed and labeled, and 20. mu.L of the sample mixture was added to each empty well.
d) Inserting the sample plate into the sample inlet of the cell counter, selecting 'AOPI cell viability', inputting sample information, and clicking 'start' to automatically measure.
According to the above procedures, the cell counts and viability of the cell solutions obtained in example 1 and example 2 were measured, and the cell counts and cell viability of PBMCs were counted, and the results are shown in Table 1 below.
Table 1: cell count and viability assay results of example 1 and example 2
Statistical results show that the total number and concentration of PBMC cells obtained by the method of example 1 are significantly higher than those obtained by the method of example 2, and both methods have higher cell viability rates.
According to the extraction method, the whole blood sample is divided into at least 2 parts, and after extraction treatment is respectively carried out, the whole blood sample is lifted and then merged, so that the influence of operation on the extraction result is reduced. The control of the centrifugation conditions in step S2 makes the mixed solution easier to separate into layers between solutions of different densities, and avoids excessive loss of lymphocytes. The control of the centrifugal force in step S5 avoids the risk of cell lysis. Compared with the existing method, the method has higher cell yield and repeatability.
Claims (6)
1. A PBMC extraction method is characterized in that: comprises the following steps of (a) carrying out,
s1: adding a whole blood sample into a centrifuge tube, adding an HBSS solution, and performing 1: 1, diluting;
s2: slowly adding the diluted blood sample into a centrifuge tube filled with lymphocyte separation liquid to ensure that the blood sample is on the upper layer of the lymphocyte separation liquid;
s3: placing the solution obtained in the step S2 at room temperature, performing horizontal centrifugal separation to layer the solution, and sequentially including a red blood cell layer, a separation liquid layer, a leukocyte layer (mononuclear cell layer), and a diluted plasma layer from bottom to top;
s4: taking a white film layer, adding a cleaning solution, and cleaning;
s5: centrifuging the mixed solution obtained in the step S4, removing supernatant, adding a cleaning solution, and resuspending cells to obtain a cell suspension;
s6: taking the cell suspension, and carrying out cell counting and activity detection.
2. The method for extracting PBMCs according to claim 1, wherein: in step S1, the sample after dilution of whole blood is divided into at least 2 parts, and extraction processes are respectively performed, and finally obtained extraction solutions are combined.
3. The method for extracting PBMCs according to claim 1, wherein: the lymphocyte separation solution is a Ficoll-Paque solution.
4. The method for extracting PBMCs according to claim 1, wherein: the conditions of the horizontal centrifugation in step S3 are to cancel the braking mode, rotate at 790g, and centrifuge for 20 minutes.
5. The method for extracting PBMCs according to claim 1, wherein: the cleaning solution in step S4 is HBSS solution.
6. The method for extracting PBMCs according to claim 1, wherein: the conditions of the centrifugation in the step S5 are to set the maximum acceleration and braking mode, the room temperature is 290g, and the centrifugation is performed for 10 minutes.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2011024575A (en) * | 2009-07-01 | 2011-02-10 | Takara Bio Inc | Method for separating cell |
| CN105255829A (en) * | 2015-11-10 | 2016-01-20 | 广州赛莱拉干细胞科技股份有限公司 | Method for separating PBMC (peripheral blood mononuclear cell) |
| CN106119199A (en) * | 2016-06-24 | 2016-11-16 | 安徽未名细胞治疗有限公司 | A kind of method of quick obtaining peripheral blood PBMC |
| CN107254438A (en) * | 2017-08-16 | 2017-10-17 | 妙顺（上海）生物科技有限公司 | The separation method of PMNC |
| US20190194614A1 (en) * | 2016-09-09 | 2019-06-27 | Yoshikazu Yonemitsu | A method for preparing mononuclear cells |
| CN110551686A (en) * | 2019-09-05 | 2019-12-10 | 广东唯泰生物科技有限公司 | method for separating peripheral blood mononuclear cells |
| CN111527395A (en) * | 2018-12-01 | 2020-08-11 | 铭道创新（北京）医疗技术有限公司 | Flow cytometry detection method for lymphocytes in immune cells |
| CN112458051A (en) * | 2020-11-11 | 2021-03-09 | 海南优尼科尔生物科技有限公司 | Extraction and collection method of peripheral blood mononuclear cells |
| US20220018745A1 (en) * | 2018-12-01 | 2022-01-20 | Mingdao Innovation (Beijing) Medical-Tech Co., Ltd. | A method for preparing lymphocyte sample for flow cytometry analysis |
-
2022
- 2022-02-07 CN CN202210115778.1A patent/CN114410580A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2011024575A (en) * | 2009-07-01 | 2011-02-10 | Takara Bio Inc | Method for separating cell |
| CN105255829A (en) * | 2015-11-10 | 2016-01-20 | 广州赛莱拉干细胞科技股份有限公司 | Method for separating PBMC (peripheral blood mononuclear cell) |
| CN106119199A (en) * | 2016-06-24 | 2016-11-16 | 安徽未名细胞治疗有限公司 | A kind of method of quick obtaining peripheral blood PBMC |
| US20190194614A1 (en) * | 2016-09-09 | 2019-06-27 | Yoshikazu Yonemitsu | A method for preparing mononuclear cells |
| CN107254438A (en) * | 2017-08-16 | 2017-10-17 | 妙顺（上海）生物科技有限公司 | The separation method of PMNC |
| CN111527395A (en) * | 2018-12-01 | 2020-08-11 | 铭道创新（北京）医疗技术有限公司 | Flow cytometry detection method for lymphocytes in immune cells |
| US20220018745A1 (en) * | 2018-12-01 | 2022-01-20 | Mingdao Innovation (Beijing) Medical-Tech Co., Ltd. | A method for preparing lymphocyte sample for flow cytometry analysis |
| CN110551686A (en) * | 2019-09-05 | 2019-12-10 | 广东唯泰生物科技有限公司 | method for separating peripheral blood mononuclear cells |
| CN112458051A (en) * | 2020-11-11 | 2021-03-09 | 海南优尼科尔生物科技有限公司 | Extraction and collection method of peripheral blood mononuclear cells |
Non-Patent Citations (6)
| Title |
|---|
| CHUANG C 等: "Enriched Peripheral Blood-Derived Mononuclear Cells for Treating Knee Osteoarthritis", 《CELL TRANSPLANT》, vol. 32, pages 1 - 11 * |
| JIA Y 等: "A Modified Ficoll-Paque Gradient Method for Isolating Mononuclear Cells from the Peripheral and Umbilical Cord Blood of Humans for Biobanks and Clinical Laboratories", 《BIOPRESERV BIOBANK》, vol. 2019, no. 2, pages 82 - 91 * |
| 刘文辉 等主编: "《免疫学检验（供医学检验技术专业使用》", vol. 2019, 31 December 2019, 中国医药科技出版社, pages: 248 - 249 * |
| 张素华 等: "Ficoll密度梯度离心法分离猪外周血单个核细胞条件的探讨", 《中国血吸虫病防治杂志》, no. 3, pages 192 - 195 * |
| 李飞 等: "人外周血单个核细胞分离优化", 《生物化工》, vol. 5, no. 5, pages 68 - 70 * |
| 蔡敏敏 等: "分离外周血单个核细胞的条件优化", 《国际检验医学杂志》, vol. 37, no. 1, pages 1 - 2 * |
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