Mesenchymal stem cell biological efficacy evaluation method and system

Abstract

The invention discloses a mesenchymal stem cell biological efficacy evaluation method and system, and relates to the technical field of mesenchymal stem cell detection.The mesenchymal stem cell biological efficacy evaluation method comprises the steps that detection results under the cell biological attribute dimension, the safety dimension, the immune regulation efficacy dimension, the anti-aging efficacy dimension and the sport injury repair efficacy dimension are obtained, each index is graded, and the evaluation result is obtained; obtaining a single score of each dimension; when the single scores of the cell biological attribute dimension and the safety dimension are not zero, determining recommended indications of the single cell strain according to the comparison of the single scores of the three efficacy dimensions of the single cell strain; and calculating the total score of each cell strain under the cell biological attribute dimension, the safety dimension and the current efficacy dimension, and screening the optimal mesenchymal stem cell strain from the current efficacy dimension according to the total score and the single score under the current efficacy dimension. Multi-dimensional indexes are considered, and the indications of the mesenchymal stem cells are recommended through a multi-level scoring strategy, so that the cell application efficiency is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of mesenchymal stem cell detection, and particularly to a method and system for evaluating the biological potency of mesenchymal stem cells. Background Art

[0002] Mesenchymal Stem Cells (MSC) can be isolated and prepared from tissues such as bone marrow, adipose tissue, synovium, bone, muscle, lung, liver, pancreas, etc., as well as amniotic fluid and umbilical cord blood. Due to the multi-directional differentiation potential, immunomodulatory and tissue repair functions of mesenchymal stem cells, and their low immunogenicity, they have currently been used in the treatment of various diseases.

[0003] However, many clinical studies have shown that due to the heterogeneity of mesenchymal stem cells caused by tissue sources, isolation methods, culture conditions, etc., the corresponding functions are presented differently, resulting in significant differences in the therapeutic effects of mesenchymal stem cells. Therefore, the selection of mesenchymal stem cell indications, treatment plans, and the heterogeneity of mesenchymal stem cells are important influencing factors. Currently, there is no reference method for screening mesenchymal stem cells suitable for specific indications based on this heterogeneity.

[0004] Biological effectiveness is an important content in the comprehensive quality evaluation of MSC products in the pre-clinical research stage and is an important basis for predicting or ensuring the clinical treatment effectiveness of related products. Traditional quality evaluation methods only focus on conventional cell markers (such as cell surface molecules CD90, CD105, etc.) and basic activities, or only use a single index to judge biological potency (such as cell surface molecules CD155, CD276, etc.), ignoring key functional indicators such as immunomodulation and repair factor expression and their mechanisms of action on specific indications, ignoring cell safety and functional efficacy under different indications for evaluation, not evaluating and screening mesenchymal stem cells separately under different indications, not establishing a mesenchymal stem cell evaluation and screening strategy for different indications, and it is difficult to quickly judge the clinical applicability of MSC. Summary of the Invention

[0005] In order to solve the above problems, the present invention proposes a method and system for evaluating the biological potency of mesenchymal stem cells, considering multi-dimensional key indicators, and realizing the recommendation of mesenchymal stem cell indications through a multi-level scoring strategy, so as to improve the cell application efficiency.

[0006] To achieve the above object, the present invention adopts the following technical solutions: In a first aspect, the present invention provides a method for evaluating the biological potency of mesenchymal stem cells, including: Obtain the detection results of a single strain of mesenchymal stem cells in the dimensions of cell biological properties, safety, immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy, and score each indicator under each dimension based on this, thereby obtaining the single-item score under each dimension; When evaluating a single strain of mesenchymal stem cells, when the single-item scores in the dimensions of cell biological properties and safety are not zero, determine the recommended indications of the single strain of mesenchymal stem cells according to the comparison results of the single-item scores in the dimensions of immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy of the single strain of mesenchymal stem cells; When evaluating multiple strains of mesenchymal stem cells under each efficacy dimension, when the single-item scores in the dimensions of cell biological properties and safety are not zero, calculate the total score of each strain of mesenchymal stem cells in the dimensions of cell biological properties, safety, and the current efficacy dimension, and screen out the best mesenchymal stem cells in the current efficacy dimension according to the total score and the single-item score in the current efficacy dimension.

[0007] As an alternative implementation, the indicators under the dimension of cell biological properties include: cell morphology, percentage of CD90 positive cells, percentage of CD105 positive cells, percentage of CD73 positive cells, percentage of CD44 positive cells, percentage of CD34 positive cells, percentage of HLA-DR positive cells, percentage of CD45 positive cells, cell viability, cell population doubling time, efficiency of fibroblast-like cell population formation, osteogenic differentiation detection, adipogenic differentiation detection, and chondrogenic differentiation detection.

[0008] As an alternative implementation, the indicators under the safety dimension include microbial safety indicators and gene safety indicators; Among them, the microbial safety indicators include: aerobic bacteria, anaerobic bacteria, mycoplasma, fungi, donor virus detection, cell virus detection, and endotoxin; The gene safety indicators include: karyotype detection, STR detection, high-incidence tumor susceptibility gene detection, cardiovascular disease susceptibility gene detection, and chronic disease susceptibility gene detection.

[0009] As an alternative implementation, the indicators under the immunomodulatory efficacy dimension include: percentage of CD276 positive cells, percentage of CD317 positive cells, percentage of CD274 positive cells, percentage of CD200 positive cells, percentage of CD146 positive cells, percentage of CD106 positive cells, HGF concentration value, TGF-β1 concentration value, GM-CSF concentration value, M-CSF concentration value, PGE2 concentration value, IDO concentration value, HLA-G5 concentration value, TSG-6 concentration value, RANTES concentration value, SDF-1 concentration value, MCP-1 concentration value, and MIP-1α concentration value.

[0010] As an alternative embodiment, the indicators in the anti-aging efficacy dimension include: the percentage value of CD142 positive, the percentage value of CD146 positive, the percentage value of CD317 positive, the percentage value of SSEA4 positive, the percentage value of CXCR4 positive, the concentration value of PGE2, the concentration value of Ang-2, the concentration value of VEGF, the concentration value of TGF-β1, the concentration value of EGF, the concentration value of PDGF-AA, the concentration value of PDGF-BB, the concentration value of HGF, the concentration value of NGF, the concentration value of SDF-1, the concentration value of ENA-78, and the concentration value of MIP-2.

[0011] As an alternative embodiment, the indicators in the exercise injury repair efficacy dimension include: the relative expression level value of BSP2, the relative expression level value of PPAR-γ, the relative expression level value of SOX9, the percentage value of CD142 positive, the percentage value of CD106 positive, the percentage value of CD317 positive, the percentage value of CXCR4 positive, the concentration value of HGF, the concentration value of VEGF, the concentration value of FGF2, the concentration value of PDGF-AA, the concentration value of IGF-1, the concentration value of Ang-2, the concentration value of MCP-1, the concentration value of GROα, the concentration value of ENA-78, the concentration value of GCP-2, the concentration value of SDF-1, the concentration value of M-CSF, and the concentration value of G-CSF.

[0012] As an alternative embodiment, the single-item score for each dimension is: ; where is the single-item score for the th dimension; is the number of evaluation indicators in the th dimension; is the weight coefficient of the th evaluation indicator in the current dimension, and the weight coefficient ranges from 0 to 1, and the sum of the weight coefficients of all evaluation indicators in a single dimension is 1; is the original score of the th evaluation indicator in the th dimension, and in the cell biological property dimension and the safety dimension, if is 0, then or is 0.

[0013] As an alternative embodiment, when evaluating a single mesenchymal stem cell line, when the single-item scores in the cell biological property dimension and the safety dimension are not zero, the efficacy dimension with the maximum single-item score among the immune regulation efficacy dimension, the anti-aging efficacy dimension, and the exercise injury repair efficacy dimension is used as the recommended indication for the single mesenchymal stem cell line.

[0014] As an alternative embodiment, when evaluating multiple mesenchymal stem cell lines under each efficacy dimension, specifically: For the immune regulation efficacy dimension, when the single-item scores in the cell biological property dimension and the safety dimension are both non-zero, calculate the total score of each mesenchymal stem cell line under the cell biological property dimension, the safety dimension, and the immune regulation efficacy dimension. First, take the mesenchymal stem cell line with the highest single-item score in the immune regulation efficacy dimension as the best mesenchymal stem cell line under the immune regulation efficacy dimension. If the single-item scores in the immune regulation efficacy dimension are the same, then take the mesenchymal stem cell line with the highest total score as the best mesenchymal stem cell line under the immune regulation efficacy dimension; For the anti-aging efficacy dimension, when the single-item scores in the cell biological property dimension and the safety dimension are both non-zero, calculate the total score of each mesenchymal stem cell line under the cell biological property dimension, the safety dimension, and the anti-aging efficacy dimension. First, take the mesenchymal stem cell line with the highest single-item score in the anti-aging efficacy dimension as the best mesenchymal stem cell line under the anti-aging efficacy dimension. If the single-item scores in the anti-aging efficacy dimension are the same, then take the mesenchymal stem cell line with the highest total score as the best mesenchymal stem cell line under the anti-aging efficacy dimension; For the sports injury repair efficacy dimension, when the single-item scores in the cell biological property dimension and the safety dimension are both non-zero, calculate the total score of each mesenchymal stem cell line under the cell biological property dimension, the safety dimension, and the sports injury repair efficacy dimension. First, take the mesenchymal stem cell line with the highest single-item score in the sports injury repair efficacy dimension as the best mesenchymal stem cell line under the sports injury repair efficacy dimension. If the single-item scores in the sports injury repair efficacy dimension are the same, then take the mesenchymal stem cell line with the highest total score as the best mesenchymal stem cell line under the sports injury repair efficacy dimension.

[0015] In a second aspect, the present invention provides a mesenchymal stem cell biological potency evaluation system, including: A scoring module configured to obtain the detection results of a single mesenchymal stem cell line under the cell biological property dimension, the safety dimension, the immune regulation efficacy dimension, the anti-aging efficacy dimension, and the sports injury repair efficacy dimension, and score each index under each dimension based on this, thereby obtaining the single-item score under each dimension; A first evaluation module configured to, when evaluating a single mesenchymal stem cell line, when the single-item scores in the cell biological property dimension and the safety dimension are both non-zero, determine the recommended indication of the single mesenchymal stem cell line according to the comparison results of the single-item scores under the immune regulation efficacy dimension, the anti-aging efficacy dimension, and the sports injury repair efficacy dimension of the single mesenchymal stem cell line; The second evaluation module is configured to calculate the total score of each mesenchymal stem cell line in the dimensions of cell biological attributes, safety, and the current efficacy dimension when evaluating multiple mesenchymal stem cell lines under each efficacy dimension, and when the individual scores in the dimensions of cell biological attributes and safety are not zero. Then, based on the total score and the individual score in the current efficacy dimension, the best mesenchymal stem cell line is screened out for the current efficacy dimension.

[0016] In a third aspect, the present invention provides an electronic device, including a memory, a processor, and computer instructions stored on the memory and running on the processor. When the computer instructions are run by the processor, the method described in the first aspect is completed.

[0017] In a fourth aspect, the present invention provides a computer-readable storage medium for storing computer instructions. When the computer instructions are executed by a processor, the method described in the first aspect is completed.

[0018] In a fifth aspect, the present invention provides a computer program product, including a computer program. When the computer program is executed by a processor, the method described in the first aspect is implemented.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention proposes a method and system for evaluating the biological efficacy of mesenchymal stem cells. Considering multi-dimensional key indicators in the dimensions of cell biological attributes, safety, immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy, through a multi-level scoring strategy of single-index scoring, individual scoring for each dimension, and total scoring, it realizes the recommendation of single-strain mesenchymal stem cell indications and the recommendation of the best cell line for each indication, improves the efficiency of cell application, and avoids the influence of mesenchymal heterogeneity caused by different tissue sources, separation methods, culture conditions, etc. of mesenchymal stem cells on the therapeutic effect of mesenchymal stem cells, and accurately screens mesenchymal stem cells suitable for specific indications; at the same time, it also considers key functional indicators such as immunomodulation and repair factor expression, evaluates the cell safety and functional efficacy under different indications, evaluates and screens mesenchymal stem cells separately under different indications, establishes an evaluation and screening strategy for mesenchymal stem cells under different indications, realizes accurate and quantitative evaluation of the quality of mesenchymal stem cells, and can provide important data references for clinical application and other scenarios.

[0020] Advantages of additional aspects of the present invention will be partially given in the following description, partially become apparent from the following description, or be understood through the practice of the present invention. Description of the Drawings

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on the provided drawings.

[0022] Figure 1 It is a flowchart of the method for evaluating the biological efficacy of mesenchymal stem cells provided in Embodiment 1 of the present invention. Specific embodiments

[0023] The following will further illustrate the present invention in conjunction with the drawings and embodiments.

[0024] It should be noted that the following detailed description is exemplary and is intended to provide further illustration of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present invention belongs.

[0025] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments of the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that the terms "comprising" and "including" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product or device that includes a series of steps or units does not necessarily have to be limited to those steps or units clearly listed, but may include other steps or units not clearly listed or inherent to these processes, methods, products or devices.

[0026] In the case of no conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other.

[0027] Embodiment 1 This embodiment provides a method for evaluating the biological efficacy of mesenchymal stem cells (MSC Biological Validity Assessment Framework, MSC-BVA). MSC-BVA effectively differentiates and evaluates MSC cell lines, providing an efficient and accurate evaluation tool for stem cell research.

[0028] It includes: Obtain the detection results of a single mesenchymal stem cell line in the dimensions of cell biological properties, safety, immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy, and score each index under each dimension accordingly, thereby obtaining the single-item score under each dimension. When evaluating a single mesenchymal stem cell line, when the individual scores in the dimensions of cell biological properties and safety are not zero, according to the comparison results of the individual scores of the single mesenchymal stem cell line in the dimensions of immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy, the recommended indications of the single mesenchymal stem cell line are determined; When evaluating multiple mesenchymal stem cell lines under each efficacy dimension, when the individual scores in the dimensions of cell biological properties and safety are not zero, calculate the total score of each mesenchymal stem cell line in the dimensions of cell biological properties, safety, and the current efficacy dimension, and screen out the best mesenchymal stem cell line for the current efficacy dimension according to the total score and the individual score in the current efficacy dimension.

[0029] First, it should be explained that the index in the form of "CD + digit N" in the following indexes represents the Nth cell surface molecule in the Cluster of Differentiation (CD) antigen system. For example, CD90 represents Cluster of Differentiation 90, CD73 represents Cluster of Differentiation 73, which is the 73rd cell surface molecule identified in the CD antigen system; CD34 represents Cluster of Differentiation 34, which is the 34th numbered in the CD antigen system, so it is called CD34; the same applies to others, and these indexes are all well-known technical terms in the industry, and each index will not be elaborated in this embodiment.

[0030] In this embodiment, the indexes under the dimension of cell biological properties include: cell morphology (normal / abnormal), CD90 positive ratio value, CD105 positive ratio value, CD73 positive ratio value, CD44 positive ratio value, CD34 positive ratio value, HLA-DR (a subtype of human leukocyte antigen (HLA) class II molecule) positive ratio value, CD45 positive ratio value, cell viability (ratio value of live cells), cell population doubling time (duration value), fibroblast-like cell population formation efficiency (efficiency value), osteogenic differentiation detection (formation / no formation), adipogenic differentiation detection (formation / no formation), and chondrogenic differentiation detection (formation / no formation).

[0031] In this embodiment, the indexes under the safety dimension include microbial safety indexes and gene safety indexes.

[0032] The microbial safety indexes include: aerobic bacteria (negative / positive), anaerobic bacteria (negative / positive), mycoplasma (negative / positive), fungi (negative / positive), 10 items of donor virus detection, cell virus detection (negative / positive), and endotoxin (qualified / exceeded standard).

[0033] Among them, the 10 items of donor virus detection include: HBsAg (hepatitis B surface antigen), anti-HCV (hepatitis C antibody), syphilis antibody, anti-HIV Ⅰ / Ⅱ (antibody against human immunodeficiency virus type Ⅰ / Ⅱ), anti-CMV IgM (antibody against cytomegalovirus IgM), HBV-DNA (deoxyribonucleic acid of hepatitis B virus), HCV-RNA (ribonucleic acid of hepatitis C virus), HIV-I-RNA (ribonucleic acid of human immunodeficiency virus type-1), anti-HTLV (antibody against human T-lymphotropic virus), EB-VCA-IgM (antibody against EB virus capsid antigen IgM).

[0034] Among them, the items of cell virus detection include HCV (hepatitis C virus), HCMV (human cytomegalovirus), EBV (Epstein-Barr virus), HPV (human papillomavirus), HBV (hepatitis B virus), HIV-1 (human immunodeficiency virus type 1).

[0035] Gene safety indicators include: karyotype detection (normal / abnormal), STR (Short Tandem Repeat) detection (no pollution / abnormal), detection of 5 susceptibility genes for high-incidence tumors (high risk / low to medium risk), detection of 6 susceptibility genes for cardiovascular and cerebrovascular diseases (high risk / low to medium risk), and detection of 3 susceptibility genes for chronic diseases (high risk / low to medium risk).

[0036] Among them, the detection of 5 susceptibility genes for high-incidence tumors includes lung cancer, gastric cancer, liver cancer, colorectal cancer, and thyroid cancer; the detection of 6 susceptibility genes for cardiovascular and cerebrovascular diseases includes hypertension, coronary heart disease, atherosclerosis, cerebral infarction, cerebral hemorrhage, and myocardial infarction; the detection of 3 susceptibility genes for chronic diseases includes hyperlipidemia, gout, and hyperhomocysteinemia.

[0037] In this embodiment, the indicators under the immune regulation efficacy dimension include: the percentage value of CD276 positive, the percentage value of CD317 positive, the percentage value of CD274 positive, the percentage value of CD200 positive, the percentage value of CD146 positive, the percentage value of CD106 positive, the concentration value of HGF (hepatocyte growth factor), the concentration value of TGF-β1 (transforming growth factor-β1), the concentration value of GM-CSF (granulocyte-macrophage colony-stimulating factor), the concentration value of M-CSF (macrophage colony-stimulating factor), the concentration value of PGE2 (prostaglandin E2), the concentration value of IDO (indoleamine 2,3-dioxygenase), the concentration value of HLA-G5 (human leukocyte antigen-G5), the concentration value of TSG-6 (tumor necrosis factor-stimulated gene 6 protein), the concentration value of RANTES (regulated on activation, normal T cell expressed and secreted), the concentration value of SDF-1 (stromal cell-derived factor-1), the concentration value of MCP-1 (monocyte chemoattractant protein-1), and the concentration value of MIP-1α (macrophage inflammatory protein-1α).

[0038] This is because it mainly involves MSC subset markers related to MSC immune regulation function and cell migration, such as CD276, CD274, CD200, CD317, CD146, CD106, immunosuppressive factors IDO, PGE2, TSG6, HLA-G5, TGF-β1, tissue repair and anti-inflammatory factors HGF, TSG6, myeloid cell regulatory factors GM-CSF, M-CSF related to anti-inflammation, and cell chemotactic recruitment factors SDF-1, MCP-1, MIP-1α, RANTES.

[0039] The indicators under the anti-aging efficacy dimension include: the percentage value of CD142 positive, the percentage value of CD146 positive, the percentage value of CD317 positive, the percentage value of SSEA4 (stage-specific embryonic antigen 4) positive, the percentage value of CXCR4 (C-X-C chemokine receptor 4) positive, the concentration value of PGE2, the concentration value of Ang-2 (angiopoietin-2), the concentration value of VEGF (vascular endothelial growth factor), the concentration value of TGF-β1, the concentration value of EGF (epidermal growth factor), the concentration value of PDGF-AA (platelet-derived growth factor-AA), the concentration value of PDGF-BB (platelet-derived growth factor-BB), the concentration value of HGF, the concentration value of NGF (nerve growth factor), the concentration value of SDF-1, the concentration value of ENA-78 (epidermal neutrophil activating peptide-78), and the concentration value of MIP-2 (macrophage inflammatory protein-2).

[0040] This is because it mainly involves MSC subset markers related to anti-fibrosis and inhibition of senescence and inflammation-related phenotypes such as CD142, CD146, CD317, SSEA4, angiogenesis and tissue repair-related factors VEGF, Ang-2, HGF, anti-inflammatory and matrix remodeling-related factors PGE2, TSG6, TGF-β1, cell activation and growth factors EGF, PDGF-AA, PDGF-BB, NGF, and cell chemotaxis and recruitment factors SDF-1, ENA-78, MIP-2.

[0041] The indicators under the dimension of sports injury repair efficacy include: the relative expression quantity value of BSP2 (bone marrow stromal protein type 2), the relative expression quantity value of PPAR-γ (peroxisome proliferator-activated receptor γ), the relative expression quantity value of SOX9 (sex-determining region Y-box protein 9 type), the positive proportion value of CD142, the positive proportion value of CD106, the positive proportion value of CD317, the positive proportion value of CXCR4 (C-X-C chemokine receptor type 4, also known as CD184 (cluster of differentiation 184)), the concentration value of HGF (hepatocyte growth factor), the concentration value of VEGF (vascular endothelial growth factor), the concentration value of FGF2 (fibroblast growth factor type 2), the concentration value of PDGF-AA, the concentration value of IGF-1 (insulin-like growth factor type 1), the concentration value of Ang-2 (angiopoietin type 2), the concentration value of MCP-1 (monocyte chemoattractant protein type 1), the concentration value of GROα (growth-regulated oncogene α), the concentration value of ENA-78 (epidermal neutrophil-activating peptide-78 type), the concentration value of GCP-2 (granulocyte chemotactic protein-2 type), the concentration value of SDF-1, the concentration value of M-CSF, and the concentration value of G-CSF (granulocyte colony-stimulating factor).

[0042] These indicators involve the differentiation ability of mesenchyme, such as the expression levels of BSP2, PPAR-γ, and SOX9 during the process of directional differentiation, cell subset markers CD142, CD106, CD317 related to sports injury inflammatory regulation and immune microenvironment remodeling, inflammation resolution-related factors MCP-1, M-CSF after sports injury, angiogenesis and tissue repair and regeneration-related factors VEGF, Ang-2, FGF2, HGF during muscle or soft tissue injury repair, matrix remodeling and anti-fibrosis-related factors PDGF-AA, IGF-1, G-CSF, and cell movement and recruitment-related factors CXCR4, SDF-1, GRO-α, GCP-2.

[0043] In this embodiment, the detection results of each indicator under the above-mentioned dimensions are scored originally, and the scoring principle of each indicator includes: (1) For cell morphology: if normal, the score y = 100; if abnormal, the score y = 0.

[0044] (2) For the CD90 positive proportion value, CD105 positive proportion value, CD73 positive proportion value, and CD44 positive proportion value: ; where y is the score; x is the positive proportion value of each index.

[0045] (3) For the CD34 positive proportion value, HLA-DR positive proportion value, and CD45 positive proportion value: ; where y is the score; x is the positive proportion value of each index.

[0046] (4) For the cell viability: ; where y is the score; x is the proportion value of live cells in the cell viability index.

[0047] (5) For the cell population doubling time: ; where y is the score; x is the duration value in the cell population doubling time index.

[0048] (6) For the formation efficiency of fibroblast-like cell population: y = 100x; where y is the score; x is the efficiency value in the formation efficiency index of fibroblast-like cell population.

[0049] (7) For osteogenic differentiation detection, adipogenic differentiation detection, and chondrogenic differentiation detection: If formation occurs, the score y = 100; if no formation occurs, the score y = 0.

[0050] (8) For aerobic bacteria, anaerobic bacteria, mycoplasma, fungi, 10 donor virus detections, and cell virus detection: If negative, the score y = 100; if positive, the score y = 0.

[0051] (9) For endotoxin: If qualified, the score y = 100; if exceeding the standard, the score y = 0.

[0052] (10) For karyotype detection and STR detection: If normal, the score y = 100; if abnormal, the score y = 0.

[0053] (11) For the detection of 5 susceptibility genes for high-incidence tumors, 6 susceptibility genes for cardiovascular and cerebrovascular diseases, and 3 susceptibility genes for chronic diseases: If high-risk, the score y = 60, otherwise the score y = 100.

[0054] (12) For the CD276 positive proportion value, CD317 positive proportion value, CD274 positive proportion value, CD200 positive proportion value, CD146 positive proportion value, CD106 positive proportion value, SSEA4 positive proportion value, CXCR4 positive proportion value, and CD142 positive proportion value: ; where y is the score; x is the positive proportion value of each indicator, m is the average value of the historical sample database, and n is the database standard deviation.

[0055] It can be understood that the historical sample database can be the detection values of historical mesenchymal stem cell lines under each of the above indicators, and the content of the database is fixed within a specific time period and is not updated in real time.

[0056] (13) For the HGF concentration value, TGF-β1 concentration value, EGF concentration value, NGF concentration value, IGF-1 concentration value, Ang-2 concentration value, PDGF-AA concentration value, PDGF-BB concentration value, VEGF concentration value, FGF2 concentration value, GM-CSF concentration value, M-CSF concentration value, GROα concentration value, G-CSF concentration value, PGE2 concentration value, IDO concentration value, HLA-G5 concentration value, TSG-6 concentration value, RANTES concentration value, SDF-1 concentration value, MCP-1 concentration value, MIP-1α concentration value, ENA-78 concentration value, GCP-2 concentration value, and MIP-2 concentration value: ; where y is the score; x is the concentration value of each indicator, m is the average value of the historical sample database, and n is the database standard deviation.

[0057] (14) For the BSP2 relative expression level value, PPAR-γ relative expression level value, and SOX9 relative expression level value: ; where y is the score; x is the concentration value of each indicator, m is the average value of the historical sample database, and n is the database standard deviation.

[0058] In this embodiment, after scoring each indicator in each dimension, the single-item score in each dimension is calculated; specifically: ; where is the single-item score of the th dimension; is the number of evaluation indicators in the th dimension; is the The weight coefficient of an evaluation index in the current dimension, where the weight coefficient ranges from 0 to 1, and the sum of the weight coefficients of all evaluation indexes in a single dimension is 1; is the th original score of the th evaluation index in the th dimension. And in the safety dimension, if

[0059] Specifically: (1) The single-item score for the cell biology attribute dimension is: if is not 0, , if is 0, then is 0.

[0060] (2) The single-item score for the safety dimension is: if is not 0, ; if is 0, then is 0.

[0061] (3) The single-item score for the immune regulation efficacy dimension is: .

[0062] (4) The single-item score for the anti-aging efficacy dimension is: .

[0063] (5) The single-item score for the sports injury repair efficacy dimension is: .

[0064] This embodiment proposes a multi-level scoring method for the score of a single index, the single-item score of each dimension, and the total score; specifically: (1) If , then the current test cell line is prohibited from clinical application.

[0065] (2) If , then the current test cell line is prohibited from clinical application.

[0066] (3) For the application of a single mesenchymal stem cell line, when evaluating a single mesenchymal stem cell line: If and , compare the , , scores of the single mesenchymal stem cell line, and the efficacy dimension with the higher score is the promotion application direction of the current single mesenchymal stem cell line.

[0067] (4) When screening multiple mesenchymal stem cell lines for a specific indication: Application for diseases related to immunomodulatory efficacy: If and , first, calculate the total score of each mesenchymal stem cell line in the dimensions of cell biological properties, safety, and immunomodulatory efficacy ; Then, for multiple mesenchymal stem cell lines, compare the scores of each mesenchymal stem cell line , and select the one with the higher score as the best mesenchymal stem cell line in the dimension of immunomodulatory efficacy; if the scores are the same, then further compare the total scores , , and select the one with the higher score as the best mesenchymal stem cell line in the dimension of immunomodulatory efficacy.

[0068] Application for anti-aging efficacy: If and , first, calculate the total score of each mesenchymal stem cell line in the dimensions of cell biological properties, safety, and anti-aging efficacy ; Then, for multiple mesenchymal stem cell lines, compare the scores of each mesenchymal stem cell line , and select the one with the higher score as the best mesenchymal stem cell line in the dimension of anti-aging efficacy; if the scores are the same, then further compare the total scores , , and select the one with the higher score as the best mesenchymal stem cell line in the dimension of anti-aging efficacy.

[0069] Application for diseases related to sports injury repair efficacy: If and , first, calculate the total score of each mesenchymal stem cell line in the dimensions of cell biological properties, safety, and sports injury repair efficacy ; Then, for multiple mesenchymal stem cell lines, compare the scores of each mesenchymal stem cell line , and select the one with the higher score as the best mesenchymal stem cell line in the dimension of sports injury repair efficacy; if the scores are the same, then further compare the total scores , , and select the one with the higher score as the best mesenchymal stem cell line in the dimension of sports injury repair efficacy.

[0070] Verification Example 1 For the first mesenchymal stem cell strain to be tested, obtain the test results in the dimensions of cell biological properties, safety, immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy. After scoring each index in each dimension, obtain the single-item score in each dimension; as shown in Tables 1 - 5.

[0071] Table 1 Dimension of cell biological properties; 。

[0072] Table 2 Dimension of safety; 。

[0073] Table 3 Dimension of immunomodulatory efficacy; 。

[0074] Table 4 Dimension of anti-aging efficacy; 。

[0075] Table 5 Dimension of sports injury repair efficacy; 。

[0076] Verification Example 2 For the second mesenchymal stem cell strain to be tested, obtain the test results in the dimensions of cell biological properties, safety, immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy. After scoring each index in each dimension, obtain the single-item score in each dimension; as shown in Tables 6 - 10.

[0077] Table 6 Dimension of cell biological properties; 。

[0078] Table 7 Dimension of safety; 。

[0079] Table 8 Dimension of immunomodulatory efficacy; 。

[0080] Table 9 Dimension of anti-aging efficacy; 。

[0081] Table 10 Dimension of sports injury repair efficacy; 。

[0082] Thus, when evaluating a single mesenchymal stem cell strain: The single-item scores of each dimension of the first mesenchymal stem cell strain to be tested are respectively , , , , , and , , indicating that this cell line may be more suitable for the treatment of diseases related to anti-aging efficacy.

[0083] The single-item scores of each dimension of the second mesenchymal stem cell line to be tested are respectively , , , , , and , , indicating that this cell line may be more suitable for the treatment of diseases related to anti-aging efficacy.

[0084] When evaluating multiple mesenchymal stem cell lines in the dimension of immunomodulatory efficacy, the single-item score of the immunomodulatory efficacy dimension of Verification Example 1 is higher than that of Verification Example 2. Then, compared with Verification Example 2, Verification Example 1 may be more suitable for diseases related to immunomodulatory efficacy.

[0085] When evaluating multiple mesenchymal stem cell lines in the dimension of anti-aging efficacy, the single-item score of the anti-aging efficacy dimension of Verification Example 2 is higher than that of Verification Example 1. Then, compared with Verification Example 1, Verification Example 2 may be more suitable for diseases related to anti-aging efficacy.

[0086] When evaluating multiple mesenchymal stem cell lines in the dimension of exercise injury repair efficacy, the single-item score of the anti-aging efficacy dimension of Verification Example 2 is higher than that of Verification Example 1. Then, compared with Verification Example 1, Verification Example 2 may be more suitable for diseases related to exercise injury repair efficacy.

[0087] Example 2 This example provides a mesenchymal stem cell biological potency evaluation system, including: A scoring module, configured to obtain the test results of a single mesenchymal stem cell line in the dimensions of cell biological properties, safety, immunomodulatory efficacy, anti-aging efficacy, and exercise injury repair efficacy, and score each index under each dimension based on this, thereby obtaining the single-item score under each dimension; The first evaluation module is configured to, when evaluating a single mesenchymal stem cell line, determine the recommended indications of the single mesenchymal stem cell line according to the comparison results of the single-item scores in the immunomodulatory efficacy dimension, anti-aging efficacy dimension, and sports injury repair efficacy dimension of the single mesenchymal stem cell line when the single-item scores in the cell biological property dimension and the safety dimension are both non-zero. The second evaluation module is configured to, when evaluating multiple mesenchymal stem cell lines in each efficacy dimension, calculate the total score of each mesenchymal stem cell line in the cell biological property dimension, the safety dimension, and the current efficacy dimension when the single-item scores in the cell biological property dimension and the safety dimension are both non-zero, and screen out the best mesenchymal stem cell line in the current efficacy dimension according to the total score and the single-item score in the current efficacy dimension.

[0088] It should be noted here that the above modules correspond to the steps described in Embodiment 1, and the examples and application scenarios implemented by the above modules and the corresponding steps are the same, but are not limited to the content disclosed in the above Embodiment 1. It should be noted that the above modules can be executed in a computer system such as a set of computer executable instructions as part of the system.

[0089] In more embodiments, there is also provided: An electronic device includes a memory, a processor, and computer instructions stored on the memory and running on the processor. When the computer instructions are run by the processor, the method described in Embodiment 1 is completed. For the sake of brevity, it will not be elaborated here.

[0090] It should be understood that in this embodiment, the processor may be a central processing unit CPU, and the processor may also be other general-purpose processors, digital signal processors DSP, application-specific integrated circuits ASIC, off-the-shelf programmable gate arrays FPGA, or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc. The general-purpose processor may be a microprocessor or the processor may also be any conventional processor, etc.

[0091] The memory may include a read-only memory and a random access memory, and provide instructions and data to the processor. A part of the memory may also include a non-volatile random access memory. For example, the memory may also store information about the device type.

[0092] A computer-readable storage medium is used to store computer instructions. When the computer instructions are executed by the processor, the method described in Embodiment 1 is completed.

[0093] The method in Embodiment 1 can be directly implemented by a hardware processor, or by a combination of hardware and software modules in the processor. The software modules can be located in mature storage media in the art, such as random access memory, flash memory, read-only memory, programmable read-only memory, or electrically erasable programmable memory, registers, etc. This storage media is located in the memory, and the processor reads the information in the memory and combines its hardware to complete the steps of the above method. To avoid repetition, it will not be described in detail here.

[0094] A computer program product includes a computer program, and when the computer program is executed by a processor, it realizes the method described in Embodiment 1.

[0095] The present invention also provides at least one computer program product tangibly stored on a non-transitory computer-readable storage medium. The computer program product includes computer-executable instructions, such as instructions included in program modules, which are executed in a device on a target real or virtual processor to perform the process / method as described above. Generally, program modules include routines, programs, libraries, objects, classes, components, data structures, etc. that perform specific tasks or implement specific abstract data types. In various embodiments, the functions of program modules can be combined or divided as needed. The machine-executable instructions for program modules can be executed within local or distributed devices. In a distributed device, program modules can be located in local and remote storage media.

[0096] The computer program code for implementing the method of the present invention can be written in one or more programming languages. This computer program code can be provided to the processor of a general-purpose computer, a special-purpose computer, or other programmable data processing devices, so that when the program code is executed by the computer or other programmable data processing devices, the functions / operations specified in the flowchart and / or block diagram are implemented. The program code can be executed entirely on the computer, partially on the computer, as an independent software package, partially on the computer and partially on a remote computer, or entirely on a remote computer or server.

[0097] In the context of the present invention, the computer program code or related data can be carried by any suitable carrier so that the device, apparatus, or processor can perform the various processes and operations described above. Examples of carriers include signals, computer-readable media, etc. Examples of signals can include electrical, optical, radio, sound, or other forms of propagated signals, such as carrier waves, infrared signals, etc.

[0098] Those of ordinary skill in the art can realize that the units and algorithm steps of the examples described in conjunction with this embodiment can be implemented by electronic hardware or a combination of computer software and electronic hardware. Whether these functions are executed in a hardware or software manner depends on the specific application and design constraints of the technical solution. Professional technicians can use different methods to implement the described functions for each specific application, but such implementation should not be considered to exceed the scope of this application.

[0099] Although the specific implementation manners of the present invention have been described above in conjunction with the accompanying drawings, it is not a limitation on the protection scope of the present invention. Those skilled in the art should understand that based on the technical solution of the present invention, various modifications or deformations that can be made by those skilled in the art without creative efforts are still within the protection scope of the present invention.

Claims

1. A method for evaluating the biological potency of mesenchymal stem cells, characterized in that, Including: Obtain the test results of a single strain of mesenchymal stem cells in the dimensions of cell biological properties, safety, immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy, and score each index under each dimension based on this, thereby obtaining the single-item score under each dimension; When evaluating a single strain of mesenchymal stem cells, when the single-item scores in the dimensions of cell biological properties and safety are not zero, determine the recommended indications of the single strain of mesenchymal stem cells according to the comparison results of the single-item scores in the dimensions of immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy of the single strain of mesenchymal stem cells; When evaluating multiple strains of mesenchymal stem cells under each efficacy dimension, when the single-item scores in the dimensions of cell biological properties and safety are not zero, calculate the total score of each strain of mesenchymal stem cells in the dimensions of cell biological properties, safety, and the current efficacy dimension, and screen out the best mesenchymal stem cells in the current efficacy dimension according to the total score and the single-item score in the current efficacy dimension.

2. The method for evaluating the biological efficacy of mesenchymal stem cells according to claim 1, wherein The indicators under the dimension of cell biological properties include: cell morphology, CD90 positive proportion value, CD105 positive proportion value, CD73 positive proportion value, CD44 positive proportion value, CD34 positive proportion value, HLA-DR positive proportion value, CD45 positive proportion value, cell viability, cell population doubling time, fibroblast-like cell population formation efficiency, osteogenic differentiation detection, adipogenic differentiation detection, and chondrogenic differentiation detection; The indicators under the safety dimension include microbial safety indicators and gene safety indicators; Among them, the microbial safety indicators include: aerobic bacteria, anaerobic bacteria, mycoplasma, fungi, donor virus detection, cell virus detection, and endotoxin; The gene safety indicators include: karyotype detection, STR detection, high-incidence tumor susceptibility gene detection, cardiovascular and cerebrovascular disease susceptibility gene detection, and chronic disease susceptibility gene detection.

3. The method for evaluating the biological efficacy of mesenchymal stem cells according to claim 1, characterized in that, The indicators under the immunomodulatory efficacy dimension include: CD276 positive proportion value, CD317 positive proportion value, CD274 positive proportion value, CD200 positive proportion value, CD146 positive proportion value, CD106 positive proportion value, HGF concentration value, TGF-β1 concentration value, GM-CSF concentration value, M-CSF concentration value, PGE2 concentration value, IDO concentration value, HLA-G5 concentration value, TSG-6 concentration value, RANTES concentration value, SDF-1 concentration value, MCP-1 concentration value, and MIP-1α concentration value.

4. The method for evaluating the biological efficacy of mesenchymal stem cells according to claim 1, wherein The indicators under the anti-aging efficacy dimension include: the percentage value of CD142 positive, the percentage value of CD146 positive, the percentage value of CD317 positive, the percentage value of SSEA4 positive, the percentage value of CXCR4 positive, the PGE2 concentration value, the Ang-2 concentration value, the VEGF concentration value, the TGF-β1 concentration value, the EGF concentration value, the PDGF-AA concentration value, the PDGF-BB concentration value, the HGF concentration value, the NGF concentration value, the SDF-1 concentration value, the ENA-78 concentration value, and the MIP-2 concentration value; The indicators under the exercise injury repair efficacy dimension include: the relative expression level value of BSP2, the relative expression level value of PPAR-γ, the relative expression level value of SOX9, the percentage value of CD142 positive, the percentage value of CD106 positive, the percentage value of CD317 positive, the percentage value of CXCR4 positive, the HGF concentration value, the VEGF concentration value, the FGF2 concentration value, the PDGF-AA concentration value, the IGF-1 concentration value, the Ang-2 concentration value, the MCP-1 concentration value, the GROα concentration value, the ENA-78 concentration value, the GCP-2 concentration value, the SDF-1 concentration value, the M-CSF concentration value, and the G-CSF concentration value.

5. The method for evaluating the biological efficacy of mesenchymal stem cells according to claim 1, characterized in that, The individual score under each dimension is as follows: ; where is the individual score of the th dimension; is the number of evaluation indicators in the th dimension; is the weight coefficient of the th evaluation indicator in the current dimension. The weight coefficient ranges from 0 to 1, and the sum of the weight coefficients of all evaluation indicators in a single dimension is 1; is the original score of the th evaluation indicator in the th dimension. In the dimensions of cell biological properties and safety, if is 0, then or is 0.

6. A method for evaluating the biological efficacy of mesenchymal stem cells according to claim 1, wherein When evaluating a single mesenchymal stem cell line, when the single-item scores in the cell biological property dimension and the safety dimension are not zero, the efficacy dimension with the maximum single-item score under the immune regulation efficacy dimension, anti-aging efficacy dimension, and exercise injury repair efficacy dimension is used as the recommended indication for the single mesenchymal stem cell line; When evaluating multiple mesenchymal stem cell lines under each efficacy dimension, specifically: For the immune regulation efficacy dimension, when the single-item scores in the cell biological property dimension and the safety dimension are not zero, calculate the total scores of each mesenchymal stem cell line in the cell biological property dimension, safety dimension, and immune regulation efficacy dimension. First, the mesenchymal stem cell line with the highest single-item score in the immune regulation efficacy dimension is the best mesenchymal stem cell line under the immune regulation efficacy dimension. If the single-item scores in the immune regulation efficacy dimension are the same, then the mesenchymal stem cell line with the highest total score is the best mesenchymal stem cell line under the immune regulation efficacy dimension; For the anti-aging efficacy dimension, when the single-item scores in the cell biological property dimension and the safety dimension are not zero, calculate the total scores of each mesenchymal stem cell line in the cell biological property dimension, safety dimension, and anti-aging efficacy dimension. First, the mesenchymal stem cell line with the highest single-item score in the anti-aging efficacy dimension is the best mesenchymal stem cell line under the anti-aging efficacy dimension. If the single-item scores in the anti-aging efficacy dimension are the same, then the mesenchymal stem cell line with the highest total score is the best mesenchymal stem cell line under the anti-aging efficacy dimension; For the dimension of sports injury repair efficacy, when the individual scores in the dimensions of cell biological properties and safety are not zero, calculate the total score of each mesenchymal stem cell line under the dimensions of cell biological properties, safety, and sports injury repair efficacy. First, take the mesenchymal stem cell line with the highest individual score in the dimension of sports injury repair efficacy as the best mesenchymal stem cell line in the dimension of sports injury repair efficacy. If the individual scores in the dimension of sports injury repair efficacy are the same, take the mesenchymal stem cell line with the highest total score as the best mesenchymal stem cell line in the dimension of sports injury repair efficacy.

7. A mesenchymal stem cell biological potency evaluation system, characterized in that, Including: A scoring module configured to obtain the test results of a single mesenchymal stem cell line under the dimensions of cell biological properties, safety, immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy, and score each indicator under each dimension based on this, thereby obtaining the individual score under each dimension; A first evaluation module configured to, when evaluating a single mesenchymal stem cell line, determine the recommended indication of the single mesenchymal stem cell line according to the comparison results of the individual scores of the single mesenchymal stem cell line under the dimensions of immunomodulatory efficacy, anti-aging efficacy, and sports injury repair efficacy when the individual scores in the dimensions of cell biological properties and safety are not zero; A second evaluation module configured to, when evaluating multiple mesenchymal stem cell lines under each efficacy dimension, calculate the total score of each mesenchymal stem cell line under the dimensions of cell biological properties, safety, and the current efficacy dimension when the individual scores in the dimensions of cell biological properties and safety are not zero, and screen out the best mesenchymal stem cell line for the current efficacy dimension according to the total score and the individual score in the current efficacy dimension.

8. An electronic device, characterized in that, It includes a memory, a processor, and computer instructions stored on the memory and running on the processor. When the computer instructions are run by the processor, the method described in any one of claims 1-6 is completed.

9. A computer-readable storage medium, characterized in that, For storing computer instructions, when the computer instructions are executed by the processor, the method described in any one of claims 1-6 is completed.

10. A computer program product, characterized in that, It includes a computer program, and when the computer program is executed by the processor, the method described in any one of claims 1-6 is implemented.

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