CN114047110A - Method for detecting galactosidase marked senescent cells by using flow cytometer - Google Patents
Method for detecting galactosidase marked senescent cells by using flow cytometer Download PDFInfo
- Publication number
- CN114047110A CN114047110A CN202111145968.XA CN202111145968A CN114047110A CN 114047110 A CN114047110 A CN 114047110A CN 202111145968 A CN202111145968 A CN 202111145968A CN 114047110 A CN114047110 A CN 114047110A
- Authority
- CN
- China
- Prior art keywords
- cells
- detected
- galactosidase
- detecting
- flow cytometer
- 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.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 30
- 108010093031 Galactosidases Proteins 0.000 title claims abstract description 24
- 102000002464 Galactosidases Human genes 0.000 title claims abstract description 24
- 239000013642 negative control Substances 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 66
- 102000005936 beta-Galactosidase Human genes 0.000 claims description 7
- 108010005774 beta-Galactosidase Proteins 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 4
- 230000036542 oxidative stress Effects 0.000 claims description 4
- 210000002950 fibroblast Anatomy 0.000 claims description 3
- 230000003287 optical effect Effects 0.000 abstract description 5
- 238000001514 detection method Methods 0.000 abstract description 4
- 230000032683 aging Effects 0.000 description 12
- 239000003814 drug Substances 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 210000002889 endothelial cell Anatomy 0.000 description 3
- 239000000834 fixative Substances 0.000 description 3
- 230000009758 senescence Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- WYXFHTWROIVOIG-RGMNGODLSA-N CNC.CN(C)[C@H](C(O)=O)CCCNC(N)=N Chemical compound CNC.CN(C)[C@H](C(O)=O)CCCNC(N)=N WYXFHTWROIVOIG-RGMNGODLSA-N 0.000 description 2
- 208000005623 Carcinogenesis Diseases 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000036952 cancer formation Effects 0.000 description 2
- 231100000504 carcinogenesis Toxicity 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 210000003606 umbilical vein Anatomy 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- 108091060290 Chromatid Proteins 0.000 description 1
- 208000028399 Critical Illness Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 101710085388 N(G),N(G)-dimethylarginine dimethylaminohydrolase Proteins 0.000 description 1
- 102100035854 N(G),N(G)-dimethylarginine dimethylaminohydrolase 1 Human genes 0.000 description 1
- YDGMGEXADBMOMJ-LURJTMIESA-N N(g)-dimethylarginine Chemical compound CN(C)C(\N)=N\CCC[C@H](N)C(O)=O YDGMGEXADBMOMJ-LURJTMIESA-N 0.000 description 1
- 108010017842 Telomerase Proteins 0.000 description 1
- YDGMGEXADBMOMJ-UHFFFAOYSA-N asymmetrical dimethylarginine Natural products CN(C)C(N)=NCCCC(N)C(O)=O YDGMGEXADBMOMJ-UHFFFAOYSA-N 0.000 description 1
- 230000032677 cell aging Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000004756 chromatid Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 229940124645 emergency medicine Drugs 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical group O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 229940059442 hemicellulase Drugs 0.000 description 1
- 108010002430 hemicellulase Proteins 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000003938 response to stress Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
Landscapes
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a method for detecting galactosidase marked senescent cells by using a flow cytometer, belonging to the field of cell detection. The method comprises the following steps: (1) detecting the average fluorescence intensity of the galactosidase marked cells to be detected; (2) detecting the average fluorescence intensity of the unlabeled cells to be detected; (3) and (3) taking the unmarked cells to be detected as negative control, setting a gate, and calculating the content of the aged cells in the cells to be detected in the step (1) according to the average fluorescence intensity obtained in the step (1) and the step (2). The invention uses the flow cytometer to replace the traditional optical microscope observation method, expands the application of the flow cytometer and provides a new method for quantitatively detecting the galactosidase marked senescent cells.
Description
Technical Field
The invention belongs to the field of cell detection, and particularly relates to a method for detecting galactosidase marked senescent cells by using a flow cytometer.
Background
Aging is a phenomenon common to all organisms. In multicellular organisms, aging is marked by a constant decline in the function of various cells and tissues. One of the most prominent features of aging is the gradual loss or deterioration of function, which occurs at various levels including molecular, cellular, tissue and individual levels. In organisms with reproducible tissues, another hallmark of aging is also an increase in the level of proliferation, the most severe of which is the occurrence of cancer. Similar to other aging-related degenerative diseases, the incidence of cancer begins to increase significantly at about the mid-life of an individual. There is increasing evidence that the stress response mechanism of cellular aging is directly or indirectly linked to a variety of aging-related disorders. Recent research results (Baker DJ et al.2016natural encapsulation p16-positive cells short life span. nature 530, 184-189) suggest that cell senescence plays an important role in increasing the age-related tumorigenesis rate, and experimental results show that the level of tumorigenesis is significantly reduced after the senescent cells are removed in mice. Therefore, monitoring the aging status of cells is of great significance for the prevention and early treatment of the related diseases.
Ilongying and the like (change of active oxygen and dimethylarginine dimethylamine hydrolase-asymmetric dimethylarginine system in the process of aging human umbilical vein endothelial cells induced by high sugar, emergency medicine of Chinese critical illness, 23 rd volume 5 th stage of 2011 in 5 months) research the change of intracellular active oxygen (ROS) level and dimethylarginine dimethylamine hydrolase-asymmetric dimethylarginine (DDAH-ADMA) system in the process of accelerating aging of endothelial cells accelerated by high sugar, and the specific research method is as follows: after a culture solution with normal sugar concentration (5.5mmol/L) and a culture solution with high sugar concentration (11.0, 22.0 and 33.0mmol/L) act on human umbilical vein endothelial cells for 48 hours, the aged cells are observed under a fluorescence microscope by using a beta-galactosidase staining method, the activity of telomerase is detected by using a polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA), the intracellular ROS level is detected by using a flow cytometer, and the ADMA content and the DDAH activity in cell supernatant are detected by using a liquid chromatograph-mass spectrometer. This study uses microscopic observations that cytoplasmic blue stained senescent cells. However, observation of senescent cells under a microscope only allows qualitative analysis of senescent cells labeled with a hemicellulase, and cannot allow quantitative analysis. Therefore, a method capable of quantitatively analyzing senescent cells is highly desired.
Disclosure of Invention
The invention aims to provide a method for quantitatively detecting galactosidase marked senescent cells by using a flow cytometer.
The invention provides a method for detecting galactosidase marked senescent cells by using a flow cytometer, which comprises the following steps:
(1) detecting the average fluorescence intensity of the galactosidase-labeled cells to be detected: after incubating galactosidase and cells to be detected together, exciting fluorescence by using 488nm wavelength laser of a flow cytometer, and receiving and emitting fluorescence signals by using a first channel to obtain the average fluorescence intensity of the galactosidase marked cells to be detected;
(2) detecting the mean fluorescence intensity of the unlabeled cells to be detected: exciting fluorescence of the unmarked cells to be detected by using 488nm wavelength laser of a flow cytometer, and receiving and emitting fluorescence signals by using a first channel to obtain the average fluorescence intensity of the unmarked cells to be detected;
(3) and (3) taking the unmarked cells to be detected as negative control, setting a gate, and calculating the content of the aged cells in the cells to be detected in the step (1) according to the average fluorescence intensity obtained in the step (1) and the step (2).
Further, the senescent cells are oxidative stress agent-induced senescent cells.
Further, the oxidative stress agent is H2O2。
Further, the cells to be detected are embryonic fibroblasts.
Further, the galactosidase is beta-galactosidase.
Further, the density of the cells to be detected is (10-50) multiplied by 104one/mL.
Further, the co-incubation time is 5-30 minutes.
Further, the co-incubation time was 15 minutes.
Further, the band pass filter of the first channel can receive the emitted fluorescence in the 526 +/-29 nm wavelength range.
Further, the band pass filter of the first channel can receive the emitted fluorescence in the 526 +/-14.5 nm wavelength range.
The invention provides a method for quantitatively detecting galactosidase mark aging cells by using a flow cytometer, expands the application of the flow cytometer and provides a new method for detecting galactosidase mark aging.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1: flow cytometry analysis.
FIG. 2: the images were observed by an optical microscope.
Detailed Description
The raw materials and equipment used in the invention are known products and are obtained by purchasing commercial products.
Example 1 method for detecting galactosidase-labeled senescent cells Using flow cytometer
Detection method
1. Establishing aging cell model
a. Preparing a culture medium in advance: DMEM high-glucose medium + 10% FBS (fetal bovine serum).
b. NIH3T3 cells (mouse embryonic fibroblasts) were plated in 6-well plates at 10X 10 cells per well4Cell, 2mL medium.
c. Mixing the mixture gently, placing at 37 ℃ and 5% CO2The cell culture box was cultured for 24 hours. The method comprises the following three groups:control, low dose, high dose groups were not dosed.
d. Replacing the fresh culture medium without adding a drug to the control group; the low-dose group and the high-dose group were discarded from the medium, washed 3 times with PBS, added with 1mL of PBS, and then the low-dose group was added with 200mM of H2O2400mM H was added to the high dose group2O2After 1 hour of treatment, the treatment solution was aspirated and fresh medium was added.
e. After further culturing for 3 days, the establishment of the senescent cell model was completed.
2. Beta-galactosidase label
a. The control group without medicine, the low dose group and the high dose group are respectively divided into two groups, one group is labeled for adherence, and the other group is labeled after digestion.
b. For adherent marker chromatids: adherent cells in a 6-well plate were aspirated, the cell culture medium was washed 1 time with PBS, 1mL of β -galactosidase label fixative was added, and fixation was performed for 15 minutes at room temperature. The cell fixative was aspirated and the cells were washed 3 times with PBS for 3 minutes each. PBS was aspirated and 1mL of labeled working solution was added to each well. Incubate overnight at 37 ℃ and seal the 6-well plate with sealing or preservative film to prevent evaporation. The beta-galactosidase staining kit was purchased from Solarbio, cat #: G1580.
and then observed under an optical microscope.
c. For the post-digestion marker set: the trypsinized cells were collected by centrifugation into a 1.5mL centrifuge tube, washed 1 time with PBS, and fixed for 15 minutes at room temperature by adding 1mL of beta-galactosidase labeling fixative. While fixing, the cells can be shaken slowly on a shaker to avoid clumping of the cells. The cell immobilizate was centrifuged off, aspirated and the cells were washed 3 times for 3 minutes each with PBS. Centrifuging, removing PBS by suction, and adding 0.5-1mL of labeled working solution into each tube. Incubate overnight at 37 ℃. Wash 1 time with PBS and resuspend the cell pellet with PBS. Besides the existing non-medicated control group, a non-labeled control group is also required to be added.
And then detected by flow cytometry. The flow cytometer uses 488nm wavelength laser to excite fluorescence, and uses the first channel (526 +/-14.5 nm) to receive the emission fluorescence signal. And (3) taking the unmarked control group as a negative control, setting a gate, and quantitatively calculating the positive percentage of the galactosidase marked senescent cells in the low-dose group and the high-dose group according to the average fluorescence intensity of the cells.
Second, the detection result
TABLE 1 percent Positive in fluorescein signal by flow cytometry
Note: MFI (mean fluorescence intensity) represents the mean fluorescence intensity.
The observation results by optical microscope are shown in FIG. 2. The staining result can only be qualitatively observed by using an optical microscope, and the number of the aged cells cannot be quantitatively analyzed.
The flow cytometry results are shown in table 1 and fig. 1. It can be seen that the control group without drug had only a few senescent cells; compared with the control group without medicine, H2O2The fluorescence intensity of the low-dose group and the high-dose group was significantly enhanced, and the fluorescence intensity of the senescent cells was the highest in the high-dose group.
The experimental results show that the method provided by the invention can be used for quantitatively detecting the content of the galactosidase marked senescent cells by using a flow cytometer.
In conclusion, the invention provides a method for quantitatively detecting the galactosidase marked senescence cells by using a flow cytometer, expands the application of the flow cytometer and provides a new method for detecting the galactosidase marked senescence cells.
Claims (10)
1. The method for detecting the galactosidase marked senescent cells by using the flow cytometer is characterized by comprising the following steps: the method comprises the following steps:
(1) detecting the average fluorescence intensity of the galactosidase-labeled cells to be detected: after incubating galactosidase and cells to be detected together, exciting fluorescence by using 488nm wavelength laser of a flow cytometer, and receiving and emitting fluorescence signals by using a first channel to obtain the average fluorescence intensity of the galactosidase marked cells to be detected;
(2) detecting the mean fluorescence intensity of the unlabeled cells to be detected: exciting fluorescence of the unmarked cells to be detected by using 488nm wavelength laser of a flow cytometer, and receiving and emitting fluorescence signals by using a first channel to obtain the average fluorescence intensity of the unmarked cells to be detected;
(3) and (3) taking the unmarked cells to be detected as negative control, setting a gate, and calculating the content of the aged cells in the cells to be detected in the step (1) according to the average fluorescence intensity obtained in the step (1) and the step (2).
2. The method of claim 1, wherein: the senescent cells are oxidative stress agent-induced senescent cells.
3. The method of claim 2, wherein: the oxidative stress agent is H2O2。
4. The method of claim 1, wherein: the cells to be detected are embryonic fibroblasts.
5. The method of claim 1, wherein: the galactosidase is beta-galactosidase.
6. The method of claim 1, wherein: the density of the cells to be detected is (10-50) multiplied by 104one/mL.
7. The method of claim 1, wherein: the co-incubation time is 5-30 minutes.
8. The method of claim 7, wherein: the co-incubation time was 15 minutes.
9. The method according to any one of claims 1 to 8, wherein: the band-pass filter of the first channel can receive the emitted fluorescence in the wavelength range of 526 +/-29 nm.
10. The method of claim 9, wherein: the band-pass filter of the first channel can receive the emitted fluorescence in the wavelength range of 526 +/-14.5 nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111145968.XA CN114047110A (en) | 2021-09-28 | 2021-09-28 | Method for detecting galactosidase marked senescent cells by using flow cytometer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111145968.XA CN114047110A (en) | 2021-09-28 | 2021-09-28 | Method for detecting galactosidase marked senescent cells by using flow cytometer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114047110A true CN114047110A (en) | 2022-02-15 |
Family
ID=80204690
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111145968.XA Pending CN114047110A (en) | 2021-09-28 | 2021-09-28 | Method for detecting galactosidase marked senescent cells by using flow cytometer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114047110A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109797192A (en) * | 2019-02-25 | 2019-05-24 | 安徽古井贡酒股份有限公司 | A kind of fast quantification senile cell detection method based on senescence associated-β-galactosidase |
| CN111044726A (en) * | 2019-12-10 | 2020-04-21 | 昆明理工大学 | Method for detecting and marking cell aging and application |
| CN111157434A (en) * | 2020-01-06 | 2020-05-15 | 四川大学华西医院 | Method for detecting luciferase-labeled cells using flow cytometry |
| CN111593089A (en) * | 2019-02-20 | 2020-08-28 | 深圳市第二人民医院 | Beta-galactosidase activity detection kit and detection method |
| CN112946264A (en) * | 2021-01-29 | 2021-06-11 | 刘韬 | Sample pretreatment method and immune cell detection method for flow cytometry |
-
2021
- 2021-09-28 CN CN202111145968.XA patent/CN114047110A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111593089A (en) * | 2019-02-20 | 2020-08-28 | 深圳市第二人民医院 | Beta-galactosidase activity detection kit and detection method |
| CN109797192A (en) * | 2019-02-25 | 2019-05-24 | 安徽古井贡酒股份有限公司 | A kind of fast quantification senile cell detection method based on senescence associated-β-galactosidase |
| CN111044726A (en) * | 2019-12-10 | 2020-04-21 | 昆明理工大学 | Method for detecting and marking cell aging and application |
| CN111157434A (en) * | 2020-01-06 | 2020-05-15 | 四川大学华西医院 | Method for detecting luciferase-labeled cells using flow cytometry |
| CN112946264A (en) * | 2021-01-29 | 2021-06-11 | 刘韬 | Sample pretreatment method and immune cell detection method for flow cytometry |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Parish et al. | Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation | |
| US7501260B2 (en) | Methods for determining efficacy of chemotherapeutic agents for a patient prior to treatment | |
| EP3240891B1 (en) | Human pluripotent stem cell-based models for predictive developmental neural toxicity | |
| EP2909313B1 (en) | Production of red blood cells and platelets from stem cells | |
| US20070059821A1 (en) | Method for preparing cell cultures from biological specimens for chemotherapeutic and other assays | |
| CN104080906A (en) | Automated system for producing induced pluripotent stem cells or differentiated cells | |
| CN110608991B (en) | Cell cycle detection kit based on mass flow detection technology and detection method | |
| CN110231275A (en) | A method of based on Flow cytometry NK cell killing activity | |
| US20200339947A1 (en) | Mesenchymal stromal cells and methods for obtaining mesenchymal stromal cells from umbilical cord | |
| Kohara et al. | KLF1 mutation E325K induces cell cycle arrest in erythroid cells differentiated from congenital dyserythropoietic anemia patient-specific induced pluripotent stem cells | |
| Reichman et al. | Mouse eosinophils: identification, isolation, and functional analysis | |
| Kos et al. | Flow cytometry-based isolation of tumor-associated regulatory T cells and assessment of their suppressive potential | |
| EP1107003A1 (en) | Methods for screening compounds active on neurons | |
| CN114047110A (en) | Method for detecting galactosidase marked senescent cells by using flow cytometer | |
| Kamalia et al. | Direct flow cytometric quantification of alkaline phosphatase activity in rat bone marrow stromal cells. | |
| EP4293108A1 (en) | Method for detecting or quantifying photoaged cells, application of same, and method for preparing photoaged cells | |
| CN115044639A (en) | Screening method of anti-aging drug | |
| EP3129469B1 (en) | Cell-surface marker of early msc aging | |
| Abuduwali et al. | Autofluorescent characteristics of human periodontal ligament cells in vitro | |
| US20240011989A1 (en) | Method for identification of viruses and diagnostic kit using the same | |
| Broughton et al. | Surface Lin28A expression consistent with cellular stress parallels indicators of senescence | |
| Buzgariu et al. | Studying Stem Cell Biology in Intact and Whole-Body Regenerating Hydra by Flow Cytometry | |
| Johnson et al. | Cell Dissociation Techniques in Salamanders | |
| JPWO2018164133A1 (en) | Cell sorting method | |
| WO2005056777A1 (en) | Adhesive stem cell fraction, method of separating the same, quality management method and quality management apparatus |
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 |