A Drug Sensitivity Analysis Method and System Based on Organoid Cell Viability

Abstract

The present invention discloses a drug sensitivity analysis method and system based on organoid cell viability, the method comprising: obtaining target sample information in an organoid culture well plate; obtaining the ATP value of the organoid in the organoid culture well plate according to the target sample information; counting the ATP value of the organoid in each well of the organoid culture well plate, and calculating the cell viability value of the organoid in each well; fitting and drawing a drug half-inhibitory concentration analysis curve according to the cell viability value of the organoid in each well. The system comprises a communication module, an ATP value reading module, a drug sensitivity analysis module, and an analysis curve drawing module. The method and system provided by the present invention facilitate drug sensitivity analysis during organoid experiments, solve the problem that drug sensitivity analysis in the past could only be achieved through endpoint detection, and can continuously display drug sensitivity analysis of organoids under different gradients.

Description

Technical Field

[0001] This invention relates to the field of drug sensitivity testing technology, and in particular to a drug sensitivity analysis method and system based on organoid cell viability. Background Technology

[0002] With the deepening of biological and medical research, traditional research models are insufficient to accurately reflect the physiological phenomena of tissues and organs in the body, leading to the emergence of new research model systems. Organoids are cells derived from stem cells or organ tissues that, after being cultured in vitro in three dimensions, can differentiate and self-organize into cell clusters possessing some specific functions and structures of corresponding human organs. Organoids cultured in three dimensions contain multiple cell types, breaking through the simple physical contact between cells and forming closer intercellular biological communication. Cells influence, induce, and feedback each other, collaboratively developing and forming functional mini-organs or tissues. These organoids can be better used to simulate the development process and physiological and pathological states of organ tissues, thus showing broad application prospects in basic research and clinical diagnosis and treatment.

[0003] Drug sensitivity testing is a method to determine an individual's drug sensitivity by testing their response to a specific drug. It can predict an individual's susceptibility to certain drugs, thereby avoiding unnecessary risks and adverse reactions. Current drug sensitivity testing technologies for organoids mostly use ATP luminescence assays to detect the amount of ATP in organoid cells after drug treatment, indirectly reflecting cell viability. However, current cell viability assays for organoids are all based on endpoint detection, which cannot continuously demonstrate the drug sensitivity of organoids at different gradients. Summary of the Invention

[0004] In view of the above-mentioned shortcomings of the prior art, the present invention provides a drug sensitivity analysis method and system based on organoid cell viability, which effectively solves the problem that previous organoid drug sensitivity analysis could only be carried out through endpoint detection.

[0005] In a first aspect, the present invention provides a method for drug sensitivity analysis based on organoid cell viability, the method comprising the following steps:

[0006] S1. Obtain target sample information from organoid culture well plates;

[0007] S2. Obtain the adenosine triphosphate (ATP) value of the organoid in the organoid culture plate based on the target sample information;

[0008] S3. Calculate the ATP value of organoids in each well of the organoid culture plate and the cell viability value of organoids in each well.

[0009] S4. Fit and plot the drug half-inhibition concentration analysis curve based on the cell viability value of the organoids in each well.

[0010] Preferably, the following steps are included before step S1:

[0011] Organoids were cultured using organoid culture plates, and the viability of the organoids was measured using a detection instrument.

[0012] Preferably, each well in the organoid culture plate uses a culture medium with a different drug concentration, and both drug-treated wells and control wells are provided.

[0013] Preferably, the detection instrument is a chemiluminescence analyzer.

[0014] Preferably, step S3 specifically includes:

[0015] The ATP values ​​of the drug-treated wells and control wells in the organoid culture plate were statistically analyzed, with the ATP value of the drug-treated wells labeled as M and the ATP value of the control wells labeled as C.

[0016] The cell viability value of the organoids in each well was calculated using the formula:

[0017]

[0018] In the above formula, Cell viability represents cell viability value, M represents the ATP value of the drug-treated well, and C represents the ATP value of the control well.

[0019] Preferably, step S4 further includes:

[0020] The drug half-inhibition concentration (WIC) and slope values ​​of the drug half-inhibition concentration (HIC) analysis curve were calculated based on the cell viability values ​​of the organoids in each well and the different drug concentrations in each well.

[0021]

[0022] In the above formula, IC50 represents the half-inhibitory concentration of the drug, Slope represents the slope value, and drug concentration represents the different drug concentrations in each well.

[0023] Secondly, the present invention also provides a drug sensitivity analysis system based on organoid cell viability, the system comprising:

[0024] The communication module is used to connect to the detection instrument for detecting organoid viability and to obtain target sample information in the organoid culture well plate through the detection information of the detection instrument.

[0025] The ATP value reading module is used to obtain the ATP value of organoids in the organoid culture well plate based on the target sample information.

[0026] The drug sensitivity analysis module is used to count the ATP value of organoids in each well of the organoid culture plate and calculate the cell viability value of organoids in each well.

[0027] The analysis curve plotting module is used to fit and plot the drug half-inhibition concentration analysis curve based on the cell viability value of the organoids in each well.

[0028] This invention provides a drug sensitivity analysis method and system based on organoid cell viability, which facilitates drug sensitivity analysis during organoid experiments and solves the problem that previous drug sensitivity analysis could only be achieved through endpoint detection. It can continuously display the drug sensitivity analysis of organoids at different gradients. Attached Figure Description

[0029] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0030] Figure 1 A flowchart of a drug sensitivity analysis method based on organoid cell viability provided in an embodiment of the present invention;

[0031] Figure 2 This is a graph showing the half-inhibitory concentration (WIC) of organoids under different drug combinations in the embodiments of the present invention.

[0032] Figure 3 The diagram shows the structure of a drug sensitivity analysis system based on organoid cell viability provided in an embodiment of the present invention. Detailed Implementation

[0033] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be further described clearly and completely below with reference to the accompanying drawings of the embodiments of this invention. It should be noted that the described embodiments are merely some embodiments of this invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0034] The terms "first" and "second" used in the embodiments of this application are for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. For example, a system, product, or device that includes a series of components or units is not limited to the listed components or units, but may optionally include unlisted components or units, or may optionally include other components or units inherent to such products or devices. In the description of this application, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0035] Drug sensitivity testing is a method to determine an individual's drug sensitivity by testing their response to a specific drug. It can predict an individual's susceptibility to certain drugs, thereby avoiding unnecessary risks and adverse reactions. Current drug sensitivity testing technologies for organoids mostly use ATP luminescence assays to detect the amount of ATP in organoid cells after drug treatment, indirectly reflecting cell viability. However, current cell viability assays for organoids are all based on endpoint detection, which cannot continuously demonstrate the drug sensitivity of organoids at different gradients.

[0036] This invention provides a drug sensitivity analysis method based on organoid cell viability, which effectively solves the problem that previous organoid drug sensitivity analyses could only be performed through endpoint detection.

[0037] First, organoids were cultured using organoid culture plates, and organoid viability was measured using a chemiluminescence analyzer. Each well in the organoid culture plate contained a culture medium with a different drug concentration, and both drug-treated and control wells were included.

[0038] In this embodiment of the invention, three drug combinations were used to determine the cell viability of organoids to assess drug sensitivity, including: fluorouracil + irinotecan, fluorouracil + oxaliplatin, and fluorouracil + irinotecan + oxaliplatin. The three drug combinations were prepared into culture media with different drug concentrations and then injected into each well of an organoid culture plate.

[0039] Figure 1 This is a flowchart of a drug sensitivity analysis method based on organoid cell viability provided in an embodiment of the present invention, as shown below. Figure 1 As shown, the method includes:

[0040] Step S1: Obtain target sample information from organoid culture wells;

[0041] The communication connection is established with a chemiluminescence analyzer to detect organoid viability in organoid culture well plates, and the target sample information is obtained through the detection information of the chemiluminescence analyzer.

[0042] Step S2: Read the ATP value of organoids in the organoid culture well plate according to the target sample information;

[0043] Step S3: Calculate the ATP value of the organoids in each well of the organoid culture plate and the cell viability value of the organoids in each well; specifically including:

[0044] The ATP values ​​of the drug-treated wells and control wells in the organoid culture plate were statistically analyzed, with the ATP value of the drug-treated wells labeled as M and the ATP value of the control wells labeled as C.

[0045] The cell viability value of the organoids in each well was calculated using the formula:

[0046]

[0047] In the above formula, Cell viability represents cell viability value, M represents the ATP value of the drug-treated well, and C represents the ATP value of the control well.

[0048] Step S4: Fit and plot the drug half-inhibition concentration analysis curve based on the cell viability value of the organoids in each well. Figure 2 This is the half-inhibitory concentration analysis curve of organoids under different drug combinations in the embodiments of the present invention.

[0049] Simultaneously, based on the cell viability value of the organoids in each well and the different drug concentrations in each well, the drug half-inhibition concentration analysis curve and the slope value were calculated:

[0050]

[0051] In the above formula, IC50 represents the half-inhibitory concentration of the drug, Slope represents the slope value, and drug concentration represents the different drug concentrations in each well.

[0052] In another aspect of the embodiments of the present invention, a drug sensitivity analysis system based on organoid cell viability is also provided. Figure 3 A structural diagram of the drug sensitivity analysis system based on organoid cell viability provided in an embodiment of the present invention is shown below. Figure 3 As shown, the system includes:

[0053] The communication module 310 is used to communicate with the detection instrument for detecting organoid viability and to obtain target sample information in the organoid culture well plate through the detection information of the detection instrument.

[0054] The ATP value reading module 320 is used to obtain the ATP value of organoids in the organoid culture well plate according to the target sample information;

[0055] The drug sensitivity analysis module 330 is used to count the ATP value of organoids in each well of the organoid culture plate and calculate the cell viability value of organoids in each well.

[0056] The analysis curve plotting module 340 is used to fit and plot the drug half-inhibition concentration analysis curve based on the cell viability value of the organoid in each well.

[0057] In summary, the present invention provides a drug sensitivity analysis method and system based on organoid cell viability, which facilitates drug sensitivity analysis during organoid experiments and solves the problem that previous drug sensitivity analysis could only be achieved through endpoint detection. It can continuously display the drug sensitivity analysis of organoids at different gradients.

[0058] In this document, the term "embodiment" means that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of this application. The appearance of this phrase in various places throughout the specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment mutually exclusive with other embodiments. It will be explicitly and implicitly understood by those skilled in the art that the embodiments described herein can be combined with other embodiments.

[0059] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

[0060] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for drug sensitivity analysis based on organoid cell viability, characterized in that, The method includes the following steps: S1. Obtain target sample information from organoid culture well plates; S2. Obtain the adenosine triphosphate (ATP) value of the organoid in the organoid culture plate based on the target sample information; S3. Calculate the ATP value of organoids in each well of the organoid culture plate and the cell viability value of organoids in each well. S4. Fit and plot the drug half-inhibition concentration analysis curve based on the cell viability value of the organoid in each well; The following steps are included before step S1: Organoids were cultured using organoid culture plates, and the viability of the organoids was detected using a detection instrument. Each well in the organoid culture plate uses a culture medium with a different drug concentration, and both drug-treated wells and control wells are provided. Step S3 specifically includes: The ATP values ​​of the drug-treated wells and control wells in the organoid culture plate were statistically analyzed, with the ATP value of the drug-treated wells labeled as M and the ATP value of the control wells labeled as C. The cell viability value of the organoids in each well was calculated using the formula: In the above formula, Cell viability represents cell viability value, M represents the ATP value of the drug-treated well, and C represents the ATP value of the control well; Step S4 further includes: The drug half-inhibition concentration (WIC) and slope values ​​of the drug half-inhibition concentration (HIC) analysis curve were calculated based on the cell viability values ​​of the organoids in each well and the different drug concentrations in each well. In the above formula, IC50 represents the half-inhibitory concentration of the drug, Slope represents the slope value, and drug concentration represents the different drug concentrations in each well.

2. The drug sensitivity analysis method based on organoid cell viability according to claim 1, characterized in that, The detection instrument is a chemiluminescence analyzer.

3. A drug sensitivity analysis system based on organoid cell viability, characterized in that, The system employing the drug sensitivity analysis method based on organoid cell viability as described in claim 1 comprises: The communication module is used to connect to the detection instrument for detecting organoid viability and to obtain target sample information in the organoid culture well plate through the detection information of the detection instrument. The ATP value reading module is used to obtain the ATP value of organoids in the organoid culture well plate based on the target sample information. The drug sensitivity analysis module is used to count the ATP value of organoids in each well of the organoid culture plate and calculate the cell viability value of organoids in each well. The analysis curve plotting module is used to fit and plot the drug half-inhibition concentration analysis curve based on the cell viability value of the organoids in each well.

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