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Method for evaluating cell killing efficacy
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An evaluation method and cell technology, applied in measurement devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of inability to collect cell images, difficult to use widely, etc., and achieve intuitive results, accurate and intuitive results.
Active Publication Date: 2021-01-29
SHANGHAI RUIYU BIOTECH
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Problems solved by technology
Therefore, although flow cytometry can specifically detect the apoptosis rate of target cells, flow cytometry is a liquid flow system and cannot collect cell images. If you need to verify the accuracy of the results, you need to use a microscope or other instruments to observe corroborate
As a result, although flow cytometry analysis methods are widely used in the scientific research field, they are difficult to be widely used in the fields of medical diagnosis and biomedical industrialization that require standardized and repeatable verification.
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Embodiment 1
[0066] This embodiment provides a method for evaluating cell killing efficacy, and the specific steps are as follows:
[0067] (1) Target cell markers:
[0068] Cultivate and collect the target cells, add fluorescent dye CFSE to the target cells for incubation, label, and prepare the labeled target cells as a cell solution;
[0070] Prepare the effector cells as a cell solution;
[0071] (3) Carry out cell co-cultivation:
[0072] Add target cells and effector cells in the culture dish at the same time (as the experimental group);
[0073] Prepare the control group cells at the same time: add only the target cells and the medium to the culture dish as the target cell control group, and add only the effector cells and the medium to another culture dish as the effector cell control group;
[0074] The above-mentioned control group and experimental group are placed under the cultivation environment and cultivated;
[0075] (4) Treat all c...
Embodiment 2
[0101] This embodiment provides a method for evaluating cell killing efficacy, and the specific steps are as follows:
[0102] (1) Target cell markers:
[0103] Transfect and express the green fluorescent proteingene into the target cells, culture and collect the target cells, and make a cell solution;
[0104] (2) Prepare effector cells:
[0105] Prepare the effector cells as a cell solution;
[0106] (3) Carry out cell co-cultivation:
[0107]Add target cells and effector cells to the culture dish at the same time, and set different effect-to-target ratios, including 1:1, 1:3 and 1:6, as the experimental group;
[0108] Prepare the control group cells at the same time: add only the target cells and the medium to the culture dish as the target cell control group, and add only the effector cells and the medium to another culture dish as the effector cell control group;
[0109] The above-mentioned control group and experimental group are placed under the cultivation envir...
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Abstract
The invention provides a method for evaluating cell killing efficacy. The evaluation method comprises the following steps: (1) co-culturing a target cell carrying a fluorescencelabel A and an effectorcell, and dyeing and labeling dead cells generated after co-culture by using a fluorescencelabel B, wherein the excitation light wavelengths corresponding to the fluorescencelabel A and the fluorescence label B are different; (2) respectively using a bright field, a fluorescence channel matched with the fluorescence label A and a fluorescence channel matched with the fluorescence label B to perform microscopic imaging on the dyed and labeled cells to obtain a microscopic image; and (3) carrying out superposition synthesis analysis on the obtained microscopic image, and evaluating the cellkilling efficacy of the effector cells according to an analysis result. According to the invention, the fluorescent reagent A and the fluorescent label B are respectively used for staining and markingthe target cells and dead cells, and then fluorescence microscopic imaging and image synthesis analysis are carried out, so that the evaluation result of the cell killingefficacy can be quickly, intuitively and accurately obtained.
Description
technical field [0001] The invention belongs to the technical field of cell killingactivity detection, and in particular relates to a method for evaluating cell killing efficacy. Background technique [0002] Compared with the current biological products, immune cell therapy products have unique characteristics, such as large initial individual differences, low scale of preparation process, most of the preparations are living cell products, and the mechanism of action is not very clear. Poor consistency, limited batches, short validity periods, and poor comparability of the same type of products, etc., all of these characteristics greatly increase the difficulty of quality control research, among which the quality control of lethal efficacy is one of the difficulties. At the same time, it is also necessary to evaluate the killing ability of immune cells in the evaluation of the patient's immune level, the evaluation of the prognosis of tumor treatment, and the evaluation of...
Claims
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Application Information
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