A method and system for in vitro culture and detection of mesenchymal stem cells and hematopoietic stem cells

Abstract

The invention relates to the technical field of cell culture detection, and specifically discloses a method and system for in vitro culture detection of mesenchymal stem cells and hematopoietic stem cells. The method comprises: analyzing a cell culture environment and culture medium components, determining a reference evaluation value for cell culture detection, judging whether the culture of mesenchymal stem cells and hematopoietic stem cells is qualified, analyzing unqualified mesenchymal stem cells and hematopoietic stem cells to obtain characteristic values ​​of mesenchymal stem cells and hematopoietic stem cells, and determining an adjustment plan for the culture of unqualified mesenchymal stem cells and hematopoietic stem cells based on the characteristic values ​​of the mesenchymal stem cells and the hematopoietic stem cells. The invention solves the problems that traditional culture of mesenchymal stem cells and hematopoietic stem cells lacks comprehensive and unified culture and detection standards, and that the culture adjustment plan has insufficient adaptability, is helpful to maintain the healthy state and function of cells, and can also improve the success rate and efficiency of cell culture.

Description

Technical Field

[0001] The present invention relates to the technical field of cell culture detection, and in particular to an in vitro culture detection method and system for mesenchymal stem cells and hematopoietic stem cells. Background Art

[0002] Mesenchymal stem cells and hematopoietic stem cells are two extremely important types of stem cells that have a wide range of applications in regenerative medicine. Mesenchymal stem cells are valued for their ability to differentiate into a variety of cell types (such as bone cells, adipocytes, and chondrocytes) as well as their anti-inflammatory and immunomodulatory abilities. Hematopoietic stem cells are the cornerstone of the hematopoietic system, responsible for maintaining the continuous renewal of blood cells, and play a key role in the treatment of blood diseases, such as leukemia and other forms of bone marrow disease. When culturing mesenchymal stem cells and hematopoietic stem cells in vitro, it is extremely important to ensure that they remain in an undifferentiated state and retain their biological functions, requiring precise control of culture conditions and regular monitoring of their physiological and differentiation states. Any deviation from ideal growth conditions may affect the quality of the cells and their clinical applicability.

[0003] At present, there are still some shortcomings in the research on in vitro culture and detection of mesenchymal stem cells and hematopoietic stem cells. Specifically, the differences in culture media, growth factors and culture conditions in the traditional culture process of mesenchymal stem cells and hematopoietic stem cells lead to inconsistency in the test results. There is a lack of comprehensive and unified culture and detection standards, and the culture adjustment schemes have insufficient adaptability. In the clinical application and industrialization of stem cell therapy, standardization of culture and detection is particularly important. Inconsistent culture conditions may lead to fluctuations in the quality and performance of cell products. Due to the lack of adaptable culture adjustment schemes, when the cell culture environment or the conditions of the cells themselves change, researchers may find it difficult to respond to these changes quickly and effectively, thereby affecting the cell culture success rate and overall experimental efficiency. Summary of the invention

[0004] In view of the deficiencies in the prior art, the present invention provides a method and system for in vitro culture and detection of mesenchymal stem cells and hematopoietic stem cells, which can effectively solve the problems involved in the above-mentioned background technology.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solutions: In a first aspect, the present invention provides a method for detecting in vitro culture of mesenchymal stem cells and hematopoietic stem cells, comprising the following steps: analyzing a cell culture environment to obtain a cell culture environment characteristic value; analyzing a cell culture medium component to obtain a cell culture medium component characteristic value, wherein the cell culture medium component includes a mesenchymal stem cell culture medium component and a hematopoietic stem cell culture medium component, and the cell culture medium characteristic value includes a mesenchymal stem cell culture medium characteristic value and a hematopoietic stem cell culture medium characteristic value; determining a cell culture detection reference evaluation value based on the cell culture environment characteristic value and the cell culture medium component characteristic value, wherein the cell culture detection reference evaluation value Including reference evaluation values ​​for mesenchymal stem cell culture detection and reference evaluation values ​​for hematopoietic stem cell culture detection; detecting the culture status of mesenchymal stem cells, and judging whether the mesenchymal stem cell culture is qualified in combination with the reference evaluation values ​​for mesenchymal stem cell culture detection; detecting the culture status of hematopoietic stem cells, and judging whether the hematopoietic stem cell culture is qualified in combination with the reference evaluation values ​​for hematopoietic stem cell culture detection; analyzing unqualified mesenchymal stem cells and hematopoietic stem cells to obtain cell characteristic values, the cell characteristic values ​​including mesenchymal stem cell characteristic values ​​and hematopoietic stem cell characteristic values; determining an adjustment plan for unqualified mesenchymal stem cell and hematopoietic stem cell culture based on the mesenchymal stem cell characteristic values ​​and the hematopoietic stem cell characteristic values.

[0006] As a further method, the cell culture environment is analyzed. The specific analysis process is: obtaining a cell culture environment data set, the cell culture environment data set specifically includes the cell culture environment temperature, the cell culture environment carbon dioxide concentration, and the cell culture environment humidity; based on the obtained cell culture environment data set, a comprehensive analysis is performed to obtain cell culture environment characteristic values, and the cell culture environment characteristic values ​​are used as an analysis basis for determining a reference evaluation value for cell culture detection.

[0007] As a further method, the components of the cell culture medium are analyzed to obtain characteristic values ​​of the cell culture medium components. The specific analysis process is as follows: analyzing the components of the mesenchymal stem cell culture medium to obtain characteristic values ​​of the mesenchymal stem cell culture medium; obtaining a mesenchymal stem cell culture medium component data set, the mesenchymal stem cell culture medium component data set specifically includes fetal bovine serum concentration, penicillin concentration, ampicillin concentration, and basic fibroblast growth factor concentration; based on the obtained mesenchymal stem cell culture medium component data set, a comprehensive analysis is performed to obtain characteristic values ​​of the mesenchymal stem cell culture medium, and the characteristic values ​​of the mesenchymal stem cell culture medium are used as an analysis basis for determining a reference evaluation value for mesenchymal stem cell culture detection; analyzing the components of the hematopoietic stem cell culture medium to obtain characteristic values ​​of the hematopoietic stem cell culture medium; obtaining a hematopoietic stem cell culture medium component data set, the hematopoietic stem cell culture medium component data set specifically includes human serum albumin concentration, stem cell factor concentration, interleukin concentration, and erythropoietin concentration; based on the obtained hematopoietic stem cell culture medium component data set, a comprehensive analysis is performed to obtain characteristic values ​​of the hematopoietic stem cell culture medium, and the characteristic values ​​of the hematopoietic stem cell culture medium are used as an analysis basis for determining a reference evaluation value for hematopoietic stem cell culture detection.

[0008] As a further method, the characteristic value of the cell culture medium component, the specific analysis process is:

[0009]

[0010] Wherein, γ1 is the characteristic value of mesenchymal stem cell culture medium, γ2 is the characteristic value of hematopoietic stem cell culture medium, tn is the fetal bovine serum concentration, qm is the penicillin concentration, xl is the ampicillin concentration, jx is the basic fibroblast growth factor concentration, rx is the human serum albumin concentration, gx is the stem cell factor concentration, gs is the interleukin concentration, ch is the erythropoietin concentration, ε1 is the compensation factor of the set tn, ε2 is the compensation factor of the set qm, ε3 is the compensation factor of the set xl, ε4 is the compensation factor of the set jx, ε5 is the compensation factor of the set rx, ε6 is the compensation factor of the set gx, ε7 is the compensation factor of the set gs, and ε8 is the compensation factor of the set ch.

[0011] As a further method, a cell culture detection reference evaluation value is determined based on the cell culture environment characteristic value and the cell culture medium component characteristic value. The specific analysis process is: determining the mesenchymal stem cell culture detection reference evaluation value based on the cell culture environment characteristic value and the mesenchymal stem cell culture medium characteristic value; storing the cell culture environment characteristic value and the mesenchymal stem cell culture medium characteristic value as a designated label, comparing the designated label with the mesenchymal stem cell culture detection reference evaluation value corresponding to each designated label stored in a database, and obtaining the mesenchymal stem cell culture detection reference evaluation value corresponding to the designated label; determining the hematopoietic stem cell culture detection reference evaluation value based on the cell culture environment characteristic value and the hematopoietic stem cell culture medium characteristic value; storing the cell culture environment characteristic value and the hematopoietic stem cell culture medium characteristic value as a designated label, comparing the designated label with the hematopoietic stem cell culture detection reference evaluation value corresponding to each designated label stored in a database, and obtaining the hematopoietic stem cell culture detection reference evaluation value corresponding to the designated label.

[0012] As a further method, whether the mesenchymal stem cell culture is qualified is judged, and the specific analysis process is: obtaining a mesenchymal stem cell culture status data set, the mesenchymal stem cell culture status data set specifically includes the mesenchymal stem cell proliferation rate, the mesenchymal stem cell apoptosis rate, the mesenchymal stem cell differentiation efficiency, and the proportion of mesenchymal stem cell morphological abnormalities; based on the obtained mesenchymal stem cell culture status data set, a comprehensive analysis is performed to obtain a mesenchymal stem cell culture status evaluation value, and the mesenchymal stem cell culture status evaluation value is used as an analysis basis for judging whether the mesenchymal stem cell culture is qualified; the mesenchymal stem cell culture status evaluation value is compared with a mesenchymal stem cell culture detection reference evaluation value; if the mesenchymal stem cell culture status evaluation value is not lower than the mesenchymal stem cell culture detection reference evaluation value, then the mesenchymal stem cell culture corresponding to the mesenchymal stem cell culture status evaluation value is qualified; if the mesenchymal stem cell culture status evaluation value is lower than the mesenchymal stem cell culture detection reference evaluation value, then the mesenchymal stem cell culture corresponding to the mesenchymal stem cell culture status evaluation value is unqualified.

[0013] As a further method, to determine whether the hematopoietic stem cell culture is qualified, the specific analysis process is: obtain a hematopoietic stem cell culture status data set, the hematopoietic stem cell culture status data set specifically includes the hematopoietic stem cell proliferation rate, the hematopoietic stem cell apoptosis rate, the hematopoietic stem cell differentiation efficiency, and the proportion of hematopoietic stem cell morphological abnormalities; based on the acquired hematopoietic stem cell culture status data set, a comprehensive analysis is performed to obtain a hematopoietic stem cell culture status evaluation value, and the hematopoietic stem cell culture status evaluation value is used as an analysis basis for determining whether the hematopoietic stem cell culture is qualified; the hematopoietic stem cell culture status evaluation value is compared with a hematopoietic stem cell culture detection reference evaluation value; if the hematopoietic stem cell culture status evaluation value is not lower than the hematopoietic stem cell culture detection reference evaluation value, then the hematopoietic stem cell culture corresponding to the hematopoietic stem cell culture status evaluation value is qualified; if the hematopoietic stem cell culture status evaluation value is lower than the hematopoietic stem cell culture detection reference evaluation value, then the hematopoietic stem cell culture corresponding to the hematopoietic stem cell culture status evaluation value is unqualified.

[0014] As a further method, unqualified mesenchymal stem cells and hematopoietic stem cells are analyzed, and the specific analysis process is as follows: the characteristic value of mesenchymal stem cells is obtained by analyzing unqualified mesenchymal stem cells; a data set of unqualified mesenchymal stem cells is obtained, and the data set of unqualified mesenchymal stem cells specifically includes the number of mesenchymal stem cell passages, the microbial contamination area of ​​the mesenchymal stem cell culture medium, and the bacterial contamination area of ​​the mesenchymal stem cell culture medium; based on the obtained data set of unqualified mesenchymal stem cells, a comprehensive analysis is performed to obtain the characteristic value of mesenchymal stem cells, and the characteristic value of mesenchymal stem cells is used as an analysis basis for determining the adjustment plan for unqualified mesenchymal stem cell culture; the characteristic value of hematopoietic stem cells is obtained by analyzing unqualified hematopoietic stem cells; a data set of unqualified hematopoietic stem cells is obtained, and the data set of unqualified hematopoietic stem cells specifically includes the number of hematopoietic stem cell passages, the microbial contamination area of ​​the hematopoietic stem cell culture medium, and the bacterial contamination area of ​​the hematopoietic stem cell culture medium; based on the obtained data set of unqualified hematopoietic stem cells, a comprehensive analysis is performed to obtain the characteristic value of hematopoietic stem cells, and the characteristic value of hematopoietic stem cells is used as an analysis basis for determining the adjustment plan for unqualified hematopoietic stem cell culture.

[0015] As a further method, unqualified mesenchymal stem cell and hematopoietic stem cell culture adjustment plans are determined. The specific analysis process is: comparing the mesenchymal stem cell characteristic value with the unqualified mesenchymal stem cell culture adjustment plans corresponding to each mesenchymal stem cell characteristic value stored in the database to obtain the unqualified mesenchymal stem cell culture adjustment plan corresponding to the mesenchymal stem cell characteristic value; comparing the hematopoietic stem cell characteristic value with the unqualified hematopoietic stem cell culture adjustment plans corresponding to each hematopoietic stem cell characteristic value stored in the database to obtain the unqualified hematopoietic stem cell culture adjustment plan corresponding to the hematopoietic stem cell characteristic value.

[0016] The second aspect of the present invention provides an in vitro culture detection system for mesenchymal stem cells and hematopoietic stem cells, comprising an environmental characteristic value acquisition module, a culture medium component characteristic value acquisition module, a culture detection reference evaluation value determination module, a mesenchymal stem cell culture qualified judgment module, a hematopoietic stem cell culture qualified judgment module, a cell characteristic value acquisition module and a culture adjustment scheme determination module, wherein: the environmental characteristic value acquisition module is used to analyze the cell culture environment to obtain the cell culture environment characteristic value; the culture medium component characteristic value acquisition module is used to analyze the cell culture medium components to obtain the cell culture medium component characteristic value, the cell culture medium components include mesenchymal stem cell culture medium components and hematopoietic stem cell culture medium components, and the cell culture medium characteristic values ​​include mesenchymal stem cell culture medium characteristic values ​​and hematopoietic stem cell culture medium characteristic values; the culture detection reference evaluation value determination module is used to determine the cell culture adjustment scheme based on the cell culture environment characteristic value and the cell culture medium component characteristic value. The cell culture detection reference evaluation value includes a mesenchymal stem cell culture detection reference evaluation value and a hematopoietic stem cell culture detection reference evaluation value; the mesenchymal stem cell culture qualified judgment module is used to detect the mesenchymal stem cell culture state, and judge whether the mesenchymal stem cell culture is qualified in combination with the mesenchymal stem cell culture detection reference evaluation value; the hematopoietic stem cell culture qualified judgment module is used to detect the hematopoietic stem cell culture state, and judge whether the hematopoietic stem cell culture is qualified in combination with the hematopoietic stem cell culture detection reference evaluation value; the cell characteristic value acquisition module is used to analyze the unqualified mesenchymal stem cells and hematopoietic stem cells to obtain cell characteristic values, and the cell characteristic values ​​include the mesenchymal stem cell characteristic value and the hematopoietic stem cell characteristic value; the culture adjustment plan determination module is used to determine the unqualified mesenchymal stem cell and hematopoietic stem cell culture adjustment plan based on the mesenchymal stem cell characteristic value and the hematopoietic stem cell characteristic value.

[0017] Compared with the prior art, the embodiments of the present invention have at least the following advantages or beneficial effects:

[0018] (1) The present invention provides a method and system for in vitro culture detection of mesenchymal stem cells and hematopoietic stem cells, accurately analyzes the characteristic values ​​of the cell culture environment and culture medium components, and compares them with standard evaluation values, thereby effectively monitoring and optimizing cell culture conditions, which not only helps to maintain the health and function of cells, but also improves the success rate and efficiency of cell culture, standardizes the cell culture and detection process, and timely discovers problems in the culture process by continuously monitoring and comparing the culture status with the reference evaluation value. Based on the analysis of specific cell characteristic values, a targeted adjustment plan is formulated for unqualified cell culture. The personalized adjustment strategy helps to meet the needs of different cell types and specific experimental conditions and improve the culture success rate.

[0019] (2) The present invention determines the reference evaluation value of cell culture detection through the characteristic values ​​of the cell culture environment and the characteristic values ​​of the cell culture medium components, defines the characteristic values ​​of the cell culture environment and the culture medium components, and then compares the experimental results with these preset reference evaluation values, thereby improving the standardization level of the experiment and ensuring that different experimenters can repeatedly produce consistent results under the same or similar conditions, thereby enhancing the repeatability of the experiment and providing a standard detection scheme. Setting the reference evaluation value enables researchers to identify deviations from normal conditions early in the cell culture process and optimize the cell culture conditions.

[0020] (3) The present invention determines the adjustment plan for unqualified mesenchymal stem cells and hematopoietic stem cell cultures through the characteristic values ​​of mesenchymal stem cells and hematopoietic stem cells, analyzes the characteristic values ​​of specific cells, and can accurately identify problems existing in cell culture. Through effective adjustment plans, the waste of materials and time caused by cell culture failure can be reduced, the efficiency of laboratory resource utilization can be improved, and the standardized adjustment plans can reduce the variation between different cell culture batches, thereby ensuring the consistency and reliability of cell products. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The present invention is further described using the accompanying drawings, but the embodiments in the accompanying drawings do not constitute any limitation to the present invention. A person skilled in the art can obtain other drawings based on the following drawings without creative work.

[0022] Figure 1 The figure is a schematic flow chart of the method steps of the present invention.

[0023] Figure 2 It is a schematic diagram of system module connection of the present invention. DETAILED DESCRIPTION

[0024] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0025] Reference Figure 1 As shown, the first aspect of the present invention provides a method for detecting in vitro culture of mesenchymal stem cells and hematopoietic stem cells, comprising: analyzing the cell culture environment to obtain characteristic values ​​of the cell culture environment.

[0026] The specific analysis process is as follows: obtaining a cell culture environment data set, which specifically includes the cell culture environment temperature, the cell culture environment carbon dioxide concentration, and the cell culture environment humidity; based on the obtained cell culture environment data set, a comprehensive analysis is performed to obtain cell culture environment characteristic values, which are used as the analysis basis for determining the reference evaluation values ​​for cell culture detection.

[0027] In a specific embodiment, the temperature of cell culture is usually maintained at 37°C, which can support optimal metabolic activities and growth conditions. Temperature fluctuations may affect the physiological state of cells, including cell growth rate, metabolism, protein synthesis, etc. Cell culture incubators are usually equipped with a built-in temperature control system that can accurately control and monitor temperature, including heating elements and temperature sensors (such as thermocouples or thermistors) to ensure that the temperature of the culture environment is constant. The carbon dioxide concentration in cell culture is usually maintained at about 5%. An appropriate CO2 concentration is necessary to maintain an appropriate pH of the culture medium, because most mammalian cell culture media use a carbonate buffer system. Instability in carbon dioxide levels will affect the pH of the culture medium, thereby affecting cell growth and survival. Cell culture incubators are usually equipped with a CO2 control system, including a CO2 sensor and a regulating valve to control and monitor the CO2 level in the incubator. Maintaining a high relative humidity (usually above 95%) is very important for preventing premature evaporation of the culture medium and maintaining a stable solute concentration in the culture medium. Inappropriate humidity can cause changes in the concentration of the culture medium and affect the physiological response of the cells. Cell culture incubators usually include a humidity control system, such as a humidifying disk or a steam generator, and a humidity sensor to monitor and adjust the humidity in the incubator.

[0028] It needs to be explained that the above-mentioned temperature directly affects the relative humidity in the incubator. Increased temperature will increase the evaporation rate of water, which may cause the loss of water in the culture medium, thereby increasing the humidity in the incubator. Without proper humidity control, this rapid evaporation may cause the culture medium to be too concentrated, affecting cell growth. Temperature changes will affect the solubility of carbon dioxide in the culture medium. As the temperature rises, the solubility of CO2 decreases, which may slightly affect the pH value of the culture medium. Humidity control of the cell culture incubator is essential for maintaining the carbon dioxide concentration and pressure balance in the incubator. If the humidity is too low, the air in the incubator may be drier, resulting in faster consumption of CO2, which may also indirectly affect the pH stability of the culture medium.

[0029] It should be explained that the above-mentioned precise monitoring and analysis of key parameters of the cell culture environment, such as temperature, carbon dioxide concentration, and humidity, can better understand the specific effects of these environmental factors on cell growth and function. By using the characteristic values ​​obtained from the environmental data set as reference evaluation values, standardized operating procedures can be established to reduce the variability between experiments. Standardization is the key to ensuring the repeatability and reliability of experimental results, ensuring the stability and optimization of the cell culture environment, and improving the quality of cell experiments and cell therapy products. Comprehensive analysis of cell culture environment data provides a data-driven decision support framework that can be used to continuously improve the culture process.

[0030] It should be explained that the specific analysis process of the above cell culture environment characteristic values ​​is as follows:

[0031]

[0032] Wherein, ω is the characteristic value of the cell culture environment, wd is the temperature of the cell culture environment, nd is the carbon dioxide concentration of the cell culture environment, sd is the humidity of the cell culture environment, θ1 is the compensation factor of the set wd, θ2 is the compensation factor of the set nd, and θ3 is the compensation factor of the set sd.

[0033] It should be explained that the above-mentioned cell culture environment characteristic values ​​are calculated through the cell culture environment temperature, cell culture environment carbon dioxide concentration, and cell culture environment humidity. By normalizing WD, ND, and SD, monitoring and integrating these environmental parameters, the environment in the cell culture incubator can be more accurately adjusted and controlled to ensure the optimal conditions for cell growth. This is especially important for sensitive cell types that rely on specific environmental conditions. The efficiency and success rate of cell culture depend to a large extent on the stability of environmental conditions. By real-time monitoring of environmental characteristic values, cell growth delays or deaths caused by environmental fluctuations can be reduced, thereby improving culture quality. Comprehensive environmental data provides data-based decision support, which can be used to analyze the impact of specific environmental parameters on cell growth and product quality, and support continuous improvement and optimization of production processes.

[0034] It should be explained that the compensation factors of wd, nd, and sd set above are obtained from the database. A mapping set of the historically measured cell culture environment temperature, cell culture environment carbon dioxide concentration, cell culture environment humidity and the compensation factors of wd, nd, and sd is established based on historical data to obtain the compensation factors of wd, nd, and sd corresponding to the current wd, nd, and sd.

[0035] It should be noted that ε1, ε2, ε3, ε4, ε5, ε6, ε7, ε8, σ1, σ2, σ3, σ4, τ1, τ2, τ3, τ4, μ1, μ2, μ3, μ4, μ5, and μ6 mentioned below are also obtained through a mapping set of historical data and compensation factors established in a database, that is, the corresponding compensation factors are obtained according to the current data.

[0036] The cell culture medium components are analyzed to obtain cell culture medium component characteristic values, the cell culture medium components include mesenchymal stem cell culture medium components and hematopoietic stem cell culture medium components, and the cell culture medium characteristic values ​​include mesenchymal stem cell culture medium characteristic values ​​and hematopoietic stem cell culture medium characteristic values.

[0037] The specific analysis process is as follows: analyzing the components of the mesenchymal stem cell culture medium to obtain the characteristic value of the mesenchymal stem cell culture medium; obtaining a mesenchymal stem cell culture medium component data set, which specifically includes fetal bovine serum concentration, penicillin concentration, ampicillin concentration, and basic fibroblast growth factor concentration; based on the obtained mesenchymal stem cell culture medium component data set, comprehensively analyzing to obtain the characteristic value of the mesenchymal stem cell culture medium, and using the characteristic value of the mesenchymal stem cell culture medium as the analysis basis for determining the reference evaluation value of mesenchymal stem cell culture detection; analyzing the components of the hematopoietic stem cell culture medium to obtain the characteristic value of the hematopoietic stem cell culture medium; obtaining a hematopoietic stem cell culture medium component data set, which specifically includes human serum albumin concentration, stem cell factor concentration, interleukin concentration, and erythropoietin concentration; based on the obtained hematopoietic stem cell culture medium component data set, comprehensively analyzing to obtain the characteristic value of the hematopoietic stem cell culture medium, and using the characteristic value of the hematopoietic stem cell culture medium as the analysis basis for determining the reference evaluation value of the hematopoietic stem cell culture detection.

[0038] It needs to be explained that the above-mentioned fetal bovine serum is a commonly used additive in cell culture. It is rich in growth factors, hormones and nutrients, and can promote cell growth and survival. The concentration of fetal bovine serum directly affects the efficiency of cell culture and the cell state. Penicillin is an antibiotic used to prevent bacterial contamination and protect cells from bacterial infection. The concentration of penicillin used needs to be sufficient to inhibit bacterial growth, but it cannot be toxic to the cultured cells. Ampicillin is a broad-spectrum antibiotic used with penicillin to enhance the effect of anti-bacterial contamination. Ampicillin can effectively resist certain penicillin-resistant bacteria. Basic fibroblast growth factor is a powerful cell proliferation and survival factor, which is often used to stimulate the proliferation of stem cells. Adding appropriate concentrations of basic fibroblast growth factor to cell culture can promote cell growth, especially for stem cells and precursor cells.

[0039] It needs to be explained that the above-mentioned fetal bovine serum provides rich nutrients, growth factors and additional protective proteins, which help cell proliferation and growth. Penicillin and ampicillin provide antibiotic protection to prevent bacterial infection and ensure a relatively sterile culture environment, which is essential for the healthy growth of cells. Basic fibroblast growth factor is an important growth factor that can significantly promote the proliferation of certain cell types, especially stem cells and precursor cells. Basic fibroblast growth factor is also present in low concentrations in fetal bovine serum, but in many cell culture systems, additional addition is required to achieve sufficient concentrations to ensure cell growth and maintain its undifferentiated state.

[0040] It should be explained that the above-mentioned human serum albumin (HSA) is a commonly used serum substitute, which is used in cell culture to provide necessary carrier proteins and maintain the stability of the extracellular environment, help stabilize the protein in the culture medium, and protect the cells from free radicals and other harmful substances. Stem cell factors (such as stem cell factor, SCF) are key signaling molecules for cell growth and maintenance, and are essential for supporting the survival, proliferation and function of stem cells. Interleukins (such as IL-3, IL-6) are an important class of cytokines that mainly play a role in immune system regulation and cell proliferation, especially in the growth and differentiation of hematopoietic cells. Erythropoietin (EPO) is a hormone that is mainly used to promote the production of red blood cells. In hematopoietic stem cell culture, EPO is a key factor in promoting the proliferation and differentiation of erythroid cells.

[0041] It needs to be explained that the above-mentioned human serum albumin, as a major carrier protein, provides nutritional support and helps maintain the stability of the extracellular environment. It can also carry and stabilize various growth factors, including interleukins and erythropoietin, so that they can remain active for a longer time in the culture environment. Stem cell factors (such as stem cell factor, SCF), interleukins and erythropoietin work together to provide the necessary signals to promote the growth and differentiation of hematopoietic stem cells. These factors each provide specific signals for different cell growth and differentiation pathways, and their interaction enhances the biological response of the cells.

[0042] It should be explained that by analyzing and determining the characteristic values ​​of culture medium components, researchers can more accurately predict and control cell growth and behavior, because these components directly affect the physiological and metabolic processes of cells. Precise control helps optimize cell growth conditions, thereby improving the success rate and efficiency of culture. Setting clear culture medium characteristic values ​​as reference evaluation values ​​can help establish unified culture standards. Using culture medium components that have been strictly analyzed and verified to culture cells can ensure that the quality and safety of the resulting cell products meet the usage standards.

[0043] Furthermore, the specific analysis process of the characteristic values ​​of the cell culture medium components is as follows:

[0044]

[0045] Wherein, γ1 is the characteristic value of mesenchymal stem cell culture medium, γ2 is the characteristic value of hematopoietic stem cell culture medium, tn is the fetal bovine serum concentration, qm is the penicillin concentration, xl is the ampicillin concentration, jx is the basic fibroblast growth factor concentration, rx is the human serum albumin concentration, gx is the stem cell factor concentration, gs is the interleukin concentration, ch is the erythropoietin concentration, ε1 is the compensation factor of the set tn, ε2 is the compensation factor of the set qm, ε3 is the compensation factor of the set xl, ε4 is the compensation factor of the set jx, ε5 is the compensation factor of the set rx, ε6 is the compensation factor of the set gx, ε7 is the compensation factor of the set gs, and ε8 is the compensation factor of the set ch.

[0046] It should be explained that the above-mentioned characteristic values ​​of the mesenchymal stem cell culture medium are calculated through the concentration of fetal bovine serum, penicillin, ampicillin and basic fibroblast growth factor. TN, QM, XL and JX are normalized. Accurate control and adjustment of the concentrations of these key components can provide the most suitable growth environment for mesenchymal stem cells, help maximize the growth potential and biological activity of the cells, and thus improve the success rate of experiments and treatments. Each additive has a specific function, such as fetal bovine serum provides nutrition and growth factors, antibiotics (penicillin and ampicillin) prevent bacterial contamination, and basic fibroblast growth factor supports cell proliferation and maintains an undifferentiated state. The correct ratio ensures the health and functionality of the cells, reduces cell variation and mortality, and monitors and adjusts the specific concentrations of the culture medium components to optimize the culture strategy based on the data.

[0047] It needs to be explained that the above-mentioned characteristic values ​​of hematopoietic stem cell culture medium are calculated through the concentration of human serum albumin, stem cell factor, interleukin and erythropoietin, and rx, gx, gs and ch are normalized. The components together provide the necessary nutrients and growth signals to support the growth and differentiation of hematopoietic stem cells. By accurately calculating and adjusting the concentrations of these components, an optimal growth environment can be created for hematopoietic stem cells, increasing cell vitality and growth efficiency. Appropriate culture medium components can reduce cell apoptosis caused by inappropriate growth conditions, and enhance cell functionality, such as enhancing the differentiation and regeneration ability of hematopoietic stem cells. The concentrations of standardized culture medium components can ensure the repeatability of the experiment and the consistency of culture results. Providing accurate culture medium characteristic values ​​can help researchers better understand the biological characteristics and response mechanisms of hematopoietic stem cells.

[0048] The cell culture test reference assessment values ​​are determined based on the cell culture environment characteristic values ​​and the cell culture medium component characteristic values. The cell culture test reference assessment values ​​include mesenchymal stem cell culture test reference assessment values ​​and hematopoietic stem cell culture test reference assessment values.

[0049] The specific analysis process is as follows: determining a reference evaluation value for mesenchymal stem cell culture detection based on characteristic values ​​of the cell culture environment and characteristic values ​​of the mesenchymal stem cell culture medium; storing the characteristic values ​​of the cell culture environment and characteristic values ​​of the mesenchymal stem cell culture medium as designated tags, comparing the designated tags with the reference evaluation values ​​for mesenchymal stem cell culture detection corresponding to each designated tag stored in a database, and obtaining a reference evaluation value for mesenchymal stem cell culture detection corresponding to the designated tag; determining a reference evaluation value for hematopoietic stem cell culture detection based on characteristic values ​​of the cell culture environment and characteristic values ​​of the hematopoietic stem cell culture medium; storing the characteristic values ​​of the cell culture environment and characteristic values ​​of the hematopoietic stem cell culture medium as designated tags, comparing the designated tags with the reference evaluation values ​​for hematopoietic stem cell culture detection corresponding to each designated tag stored in a database, and obtaining a reference evaluation value for hematopoietic stem cell culture detection corresponding to the designated tags.

[0050] It should be explained that the labeling of specific culture conditions and comparison with reference evaluation values ​​can ensure the consistency of culture conditions and evaluation standards between different experiments and even between different researchers, which significantly improves the repeatability and reliability of experimental results. Ensuring the high quality and consistency of the cell culture process is crucial to the safety and effectiveness of cell therapy in clinical applications. Through precise monitoring and evaluation of culture conditions, the quality of cell products can be greatly improved, and the risk of use caused by quality problems can be reduced. It provides a data-based decision support system to help researchers make more scientific and accurate decisions based on real-time and historical data.

[0051] The culture status of mesenchymal stem cells is tested, and combined with the reference evaluation value of mesenchymal stem cell culture test to determine whether the mesenchymal stem cell culture is qualified.

[0052] The specific analysis process is as follows: obtaining a mesenchymal stem cell culture status data set, which specifically includes a mesenchymal stem cell proliferation rate, a mesenchymal stem cell apoptosis rate, a mesenchymal stem cell differentiation efficiency, and a proportion of mesenchymal stem cell morphological abnormalities; based on the obtained mesenchymal stem cell culture status data set, a comprehensive analysis is performed to obtain a mesenchymal stem cell culture status evaluation value, which is used as an analysis basis for judging whether the mesenchymal stem cell culture is qualified;

[0053] The evaluation value of mesenchymal stem cell culture status is calculated as follows:

[0054]

[0055] Wherein, α is the evaluation value of the culture state of mesenchymal stem cells, zz is the proliferation rate of mesenchymal stem cells, dw is the apoptosis rate of mesenchymal stem cells, fh is the differentiation efficiency of mesenchymal stem cells, yc is the proportion of abnormal morphology of mesenchymal stem cells, σ1 is the compensation factor of the set zz, σ2 is the compensation factor of the set dw, σ3 is the compensation factor of the set fh, and σ4 is the compensation factor of the set yc;

[0056] It should be explained that the above-mentioned evaluation value of the culture status of mesenchymal stem cells is calculated through the proliferation rate of mesenchymal stem cells, the apoptosis rate of mesenchymal stem cells, the differentiation efficiency of mesenchymal stem cells, and the proportion of abnormal morphology of mesenchymal stem cells. ZZZ, DW, FH, and YC are normalized, and the proliferation rate, apoptosis rate, differentiation efficiency, and the proportion of abnormal morphology are taken into consideration together, providing a comprehensive perspective on the cell culture status. The comprehensive evaluation helps researchers to fully understand the health status and functional status of cells. Regular monitoring and evaluation of these key indicators can make precise adjustments to the culture conditions, such as changing the composition of the culture medium, adjusting the culture environment, etc., so as to optimize the growth and functional expression of cells. By real-time monitoring of these key indicators, possible problems in the culture process, such as poor cell proliferation, high apoptosis rate, or insufficient differentiation, can be discovered early, and timely measures can be taken to correct them.

[0057] The mesenchymal stem cell culture status assessment value is compared with the mesenchymal stem cell culture detection reference assessment value; if the mesenchymal stem cell culture status assessment value is not lower than the mesenchymal stem cell culture detection reference assessment value, the mesenchymal stem cell culture corresponding to the mesenchymal stem cell culture status assessment value is qualified; if the mesenchymal stem cell culture status assessment value is lower than the mesenchymal stem cell culture detection reference assessment value, the mesenchymal stem cell culture corresponding to the mesenchymal stem cell culture status assessment value is unqualified.

[0058] In a specific embodiment, by comparing the actual culture status assessment value with the standard reference assessment value, it is possible to intuitively monitor and confirm whether the cell culture process has reached the preset quality standard, and can effectively ensure that the cultured cells have the desired biological characteristics and functions, thereby ensuring the reliability of the experimental results. If the culture status assessment value is lower than the reference assessment value, this comparison mechanism can promptly reveal possible problems in cell culture, such as malnutrition, environmental control errors, pollution, etc., thereby allowing researchers to quickly take measures to adjust and optimize, which not only improves the culture efficiency but also reduces the waste of resources. Setting a specific reference assessment value and using it as a basis for comparison helps to standardize the cell culture process and ensure that different experimenters and even different laboratories can repeatedly produce consistent cell culture results, which is crucial for the repeatability of scientific research and consistency in clinical practice.

[0059] The hematopoietic stem cell culture status is tested and combined with the hematopoietic stem cell culture test reference evaluation value to determine whether the hematopoietic stem cell culture is qualified.

[0060] The specific analysis process is as follows: obtaining a hematopoietic stem cell culture status data set, which specifically includes the hematopoietic stem cell proliferation rate, hematopoietic stem cell apoptosis rate, hematopoietic stem cell differentiation efficiency, and the proportion of hematopoietic stem cell morphological abnormalities; based on the obtained hematopoietic stem cell culture status data set, a comprehensive analysis is performed to obtain a hematopoietic stem cell culture status evaluation value, which is used as an analysis basis for judging whether the hematopoietic stem cell culture is qualified;

[0061] It should be explained that the above proliferation rate refers to the growth and reproduction rate of mesenchymal stem cells, which is usually measured by the increase in the number of cells. The proliferation rate can be determined by a variety of methods, such as using a cell counter to directly count the number of cells. The apoptosis rate refers to the proportion of cell apoptosis. Cell apoptosis is a programmed cell death process that is essential for maintaining the health of tissues. Cell apoptosis can be detected by flow cytometry using specific dyes such as PI. These dyes can distinguish between living cells, early apoptotic cells, and late apoptotic or necrotic cells. The differentiation efficiency refers to the proportion of mesenchymal stem cells that are successfully converted into target cell types after being induced to differentiate into specific cell types (osteoblasts, adipocytes, or chondrocytes). The differentiation efficiency is usually evaluated by specific cell staining (such as Alizarin red staining to detect osteoblasts and Oil Red O staining to detect adipocytes) and / or by gene expression analysis (such as real-time PCR) to evaluate the expression level of specific marker genes. The morphological abnormality ratio refers to the proportion of cells with abnormal morphology in a cell population, which may include abnormalities in cell size, shape, or attachment properties.

[0062] It needs to be explained that the high proliferation rate mentioned above means that cell division is active and the cell number is increasing rapidly, while a high apoptosis rate indicates that the proportion of cell death is increasing, which will inhibit the growth of the overall cell number. The ideal situation is a high proliferation rate and a low apoptosis rate, which can maintain the healthy expansion of the cell population. A high proliferation rate may be accompanied by a low differentiation efficiency, because cells with high proliferative activity may be more likely to maintain an undifferentiated state rather than enter the differentiation process. On the contrary, inducing cell differentiation usually requires slowing down the cell proliferation rate so that the cells can perform complex differentiation programs. A high apoptosis rate may be associated with a high proportion of morphological abnormalities, because cell death caused by physiological or biochemical stress can also cause changes in cell morphology. Abnormal cell morphology may be part of the changes in cell structure and function that occur during apoptosis. During differentiation, cells usually change their morphology to adapt to new functional roles. If the differentiation is insufficient or abnormal, this may manifest as morphological abnormalities. High differentiation efficiency is usually associated with normal morphological changes, while low differentiation efficiency may be associated with an increased proportion of morphological abnormalities.

[0063] The evaluation value of hematopoietic stem cell culture status is calculated as follows:

[0064]

[0065] Wherein, β is the evaluation value of hematopoietic stem cell culture status, z is the proliferation rate of hematopoietic stem cells, d is the apoptosis rate of hematopoietic stem cells, f is the differentiation efficiency of hematopoietic stem cells, c is the proportion of hematopoietic stem cell morphological abnormalities, τ1 is the compensation factor of z, τ2 is the compensation factor of d, τ3 is the compensation factor of f, τ4 is the compensation factor of c, and e is a natural constant;

[0066] It needs to be explained that the above-mentioned hematopoietic stem cell culture status evaluation value is calculated through the hematopoietic stem cell proliferation rate, hematopoietic stem cell apoptosis rate, hematopoietic stem cell differentiation efficiency, and the proportion of hematopoietic stem cell morphological abnormalities. z, d, f, and c are normalized, and these key biological parameters are combined to comprehensively evaluate the health status and function of hematopoietic stem cells. According to the feedback of the culture status evaluation value, the culture conditions, such as temperature, CO2 concentration, culture medium components, etc., can be accurately adjusted to promote cell proliferation, reduce apoptosis, enhance differentiation efficiency or improve cell morphology. Real-time evaluation of the culture status can promptly detect problems, such as insufficient cell proliferation or excessively high apoptosis rate, so as to quickly take intervention measures to improve the overall culture success rate. By continuously monitoring and evaluating the culture status of hematopoietic stem cells, the consistency and reliability of cell products can be ensured.

[0067] Compare the hematopoietic stem cell culture status assessment value with the hematopoietic stem cell culture test reference assessment value; if the hematopoietic stem cell culture status assessment value is not lower than the hematopoietic stem cell culture test reference assessment value, the hematopoietic stem cell culture corresponding to the hematopoietic stem cell culture status assessment value is qualified; if the hematopoietic stem cell culture status assessment value is lower than the hematopoietic stem cell culture test reference assessment value, the hematopoietic stem cell culture corresponding to the hematopoietic stem cell culture status assessment value is unqualified.

[0068] In a specific embodiment, a quantitative assessment of the hematopoietic stem cell culture status is performed and compared with a set standard (reference assessment value) to ensure that the cultured cells meet predetermined quality standards, which is the basis for ensuring cell functionality and safety. If the culture status assessment value is lower than the reference assessment value, this comparison mechanism can promptly reveal possible problems in the cell culture process, such as environmental control errors, malnutrition or pollution, etc. Timely discovery of these problems allows rapid measures to be taken to adjust and optimize culture conditions, avoiding greater waste of resources and potential failures. Comparing the assessment value with a fixed reference assessment value helps to achieve standardization of the cell culture process, ensure repeatability between different experiments and between different researchers and laboratories, and improve the consistency and reliability of experimental data.

[0069] Mesenchymal stem cells and hematopoietic stem cells that fail to be cultured are analyzed to obtain cell characteristic values, which include mesenchymal stem cell characteristic values ​​and hematopoietic stem cell characteristic values.

[0070] The specific analysis process is as follows: the characteristic value of mesenchymal stem cells is obtained by analyzing the mesenchymal stem cells that fail to be cultured; a data set of mesenchymal stem cells that fail to be cultured is obtained, and the data set of mesenchymal stem cells that fail to be cultured specifically includes the number of mesenchymal stem cell passages, the microbial contamination area of ​​the mesenchymal stem cell culture medium, and the bacterial contamination area of ​​the mesenchymal stem cell culture medium; based on the obtained data set of mesenchymal stem cells that fail to be cultured, a comprehensive analysis is performed to obtain the characteristic value of mesenchymal stem cells, and the characteristic value of mesenchymal stem cells is used as the analysis basis for determining the adjustment plan for the unqualified mesenchymal stem cell culture; the characteristic value of hematopoietic stem cells is obtained by analyzing the hematopoietic stem cells that fail to be cultured; a data set of hematopoietic stem cells that fail to be cultured is obtained, and the data set of hematopoietic stem cells that fail to be cultured specifically includes the number of hematopoietic stem cell passages, the microbial contamination area of ​​the hematopoietic stem cell culture medium, and the bacterial contamination area of ​​the hematopoietic stem cell culture medium; based on the obtained data set of hematopoietic stem cells that fail to be cultured, a comprehensive analysis is performed to obtain the characteristic value of hematopoietic stem cells, and the characteristic value of hematopoietic stem cells is used as the analysis basis for determining the adjustment plan for the unqualified hematopoietic stem cell culture.

[0071] In a specific embodiment, the cell passage number refers to the maximum number of times all cells have been propagated and divided since the initial isolation and culture. The cell passage number reflects the degree of cell aging. The higher the passage number, the lower the genetic stability and biological activity of the cells may be. The microbial contamination area of ​​the cell culture medium refers to the area or proportion of the culture medium contaminated by microorganisms (such as fungi and yeast). Microbial contamination can affect the growth and function of cells and even lead to the failure of cell culture. The bacterial contamination area of ​​the cell culture medium refers to the area or proportion of the culture medium contaminated by bacteria. Bacterial contamination not only affects the healthy growth of cells, but also may cause deviations in cell experimental results.

[0072] It needs to be explained that the above-mentioned high number of passages may lead to unstable gene expression and decreased physiological function of cells, making cells more susceptible to external environment influences, including a higher risk of infection. The weakening of cell defense mechanisms may make the culture medium more susceptible to contamination by microorganisms and bacteria. High-passage cells may change the conditions of the culture medium due to changes in their physiological state, such as changes in pH, increased nutrient consumption, etc. These changes may affect the growth conditions of microorganisms and indirectly affect the degree of contamination. Microbial contamination in the culture medium (including bacteria and other microorganisms such as fungi) will compete for nutrients required by cells, produce harmful metabolites, and lead to a deterioration of the cell growth environment, directly affecting the health and survival of cells.

[0073] It should be explained that the above analysis of the specific characteristics of cells that fail the culture (such as the number of passages, microbial and bacterial contamination area) can accurately diagnose specific problems in the culture process, such as whether the cells are aging due to excessive number of passages, or whether the culture failure is caused by microbial contamination. Accurate problem diagnosis makes subsequent adjustment measures more targeted and improves processing efficiency. Based on the characteristic values ​​obtained from the analysis, specific adjustment plans can be formulated to solve the identified problems. If it is found that the problem is due to microbial contamination, aseptic operations can be strengthened and the antibiotic formulation of the culture medium can be improved. Targeted adjustments can directly solve the problem and prevent it from happening again. Timely identification and resolution of problems in the culture process can significantly improve the success rate of cell culture and the quality of cultured cells. Effective identification and resolution of problems in culture can avoid repeated experiments and excessive waste of resources.

[0074] It should be explained that the specific analysis process of the above cell characteristic values ​​is as follows:

[0075]

[0076] In the formula, δ1 is the characteristic value of mesenchymal stem cells, δ2 is the characteristic value of hematopoietic stem cells, jc is the number of mesenchymal stem cell passages, ws is the microbial contamination area of ​​mesenchymal stem cell culture medium, xj is the bacterial contamination area of ​​mesenchymal stem cell culture medium, xc is the number of hematopoietic stem cell passages, xs is the microbial contamination area of ​​hematopoietic stem cell culture medium, xx is the bacterial contamination area of ​​hematopoietic stem cell culture medium, μ1 is the compensation factor of the set jc, μ2 is the compensation factor of the set ws, μ3 is the compensation factor of the set xj, μ4 is the compensation factor of the set xc, μ5 is the compensation factor of the set xs, μ6 is the compensation factor of the set xx, and e is a natural constant.

[0077] It should be explained that the above-mentioned characteristic values ​​of mesenchymal stem cells are calculated through the number of mesenchymal stem cell passages, the microbial contamination area of ​​the mesenchymal stem cell culture medium, and the bacterial contamination area of ​​the mesenchymal stem cell culture medium. By normalizing jc, ws, and xj and analyzing these specific characteristics, the causes of unqualified culture can be accurately identified, such as whether cell aging is caused by excessive passages, or cell death is caused by microbial contamination, thereby directly solving the root cause of the problem. Timely and accurate adjustments can significantly improve the success rate of repeated culture, avoid the same mistakes from happening again, and improve the overall culture efficiency. Identifying and solving culture problems can reduce the waste of time and resources caused by repeated experiments and failed cultures.

[0078] It needs to be explained that the above-mentioned hematopoietic stem cell characteristic values ​​are calculated through the number of hematopoietic stem cell passages, the microbial contamination area of ​​hematopoietic stem cell culture medium, and the bacterial contamination area of ​​hematopoietic stem cell culture medium. xc, xs, and xx are normalized. By effectively controlling the culture conditions and adjusting unqualified cultures, the experimental results obtained can be guaranteed to be highly reliable and repeatable. The causes of unqualified culture can also be accurately identified, such as whether cell aging is caused by excessive passages, or cell death is caused by microbial contamination, thereby directly solving the root cause of the problem. Systematically analyzing and solving problems in cell culture will help to continuously improve operating procedures and protocols, set more efficient and safer standards for future experiments, and ensure that each step of culture meets quality standards. It is essential for the safety and effectiveness of clinical-grade cell therapies. By fine-tuning the culture regimen, the quality of cell products can be greatly improved.

[0079] Based on the characteristic values ​​of mesenchymal stem cells and hematopoietic stem cells, the adjustment plan for the culture of unqualified mesenchymal stem cells and hematopoietic stem cells is determined.

[0080] The specific analysis process is: comparing the mesenchymal stem cell characteristic value with the unqualified mesenchymal stem cell culture adjustment plan corresponding to each mesenchymal stem cell characteristic value stored in the database to obtain the unqualified mesenchymal stem cell culture adjustment plan corresponding to the mesenchymal stem cell characteristic value; comparing the hematopoietic stem cell characteristic value with the unqualified hematopoietic stem cell culture adjustment plan corresponding to each hematopoietic stem cell characteristic value stored in the database to obtain the unqualified hematopoietic stem cell culture adjustment plan corresponding to the hematopoietic stem cell characteristic value.

[0081] In a specific embodiment, by comparing the characteristic values ​​with known unqualified cases, the possible causes of the current culture problems can be quickly identified, and effective adjustment measures can be taken with reference to past experience, which greatly shortens the time for problem diagnosis and resolution and improves processing efficiency. The use of verified adjustment schemes to optimize unqualified culture conditions can significantly improve the success rate of cell culture. Through effective adjustment schemes, the waste of resources caused by repeated experiments and failed cultures can be reduced. In the case of high cell culture costs, this method can significantly reduce costs and improve resource utilization efficiency. Different cell batches may show different culture characteristics due to slight differences in genes or phenotypes. Through detailed characteristic value analysis and corresponding adjustment schemes, more refined and individualized culture strategies can be achieved. The data generated during the cell culture process can be systematized and databased, which can provide strong data support for cell biology research. The data-driven decision-making process can be based on empirical and historical data to improve the scientificity and effectiveness of decision-making.

[0082] Reference Figure 2 As shown, the second aspect of the present invention provides an in vitro culture detection system for mesenchymal stem cells and hematopoietic stem cells, including an environmental characteristic value acquisition module, a culture medium component characteristic value acquisition module, a culture detection reference evaluation value determination module, a mesenchymal stem cell culture qualified judgment module, a hematopoietic stem cell culture qualified judgment module, a cell characteristic value acquisition module and a culture adjustment plan determination module.

[0083] The environment characteristic value acquisition module is used to analyze the cell culture environment and obtain the cell culture environment characteristic value.

[0084] The culture medium component characteristic value acquisition module is used to analyze the cell culture medium components to obtain the cell culture medium component characteristic values. The cell culture medium components include mesenchymal stem cell culture medium components and hematopoietic stem cell culture medium components. The cell culture medium characteristic values ​​include mesenchymal stem cell culture medium characteristic values ​​and hematopoietic stem cell culture medium characteristic values.

[0085] The culture detection reference evaluation value determination module is used to determine the cell culture detection reference evaluation value based on the cell culture environment characteristic value and the cell culture medium component characteristic value. The cell culture detection reference evaluation value includes the mesenchymal stem cell culture detection reference evaluation value and the hematopoietic stem cell culture detection reference evaluation value.

[0086] The mesenchymal stem cell culture qualification judgment module is used to detect the culture status of the mesenchymal stem cells and judge whether the mesenchymal stem cell culture is qualified in combination with the mesenchymal stem cell culture detection reference evaluation value.

[0087] The hematopoietic stem cell culture qualification judgment module is used to detect the hematopoietic stem cell culture status and judge whether the hematopoietic stem cell culture is qualified in combination with the hematopoietic stem cell culture detection reference evaluation value.

[0088] The cell characteristic value acquisition module is used to analyze the mesenchymal stem cells and hematopoietic stem cells that fail to be cultured to obtain cell characteristic values, which include mesenchymal stem cell characteristic values ​​and hematopoietic stem cell characteristic values.

[0089] The culture adjustment plan determination module is used to determine unqualified mesenchymal stem cell and hematopoietic stem cell culture adjustment plans based on the characteristic values ​​of mesenchymal stem cells and hematopoietic stem cells.

[0090] The above contents are merely examples and explanations of the structure of the present invention. The technicians in this technical field may make various modifications or additions to the specific embodiments described or replace them in a similar manner. As long as they do not deviate from the structure of the invention or exceed the scope defined by the claims, they should all fall within the protection scope of the present invention.

Claims

1. A method for in vitro culture and detection of mesenchymal stem cells and hematopoietic stem cells, characterized in that: The following steps are involved: Analyze the cell culture environment to obtain characteristic values ​​of the cell culture environment; Analyzing cell culture medium components to obtain cell culture medium component characteristic values, wherein the cell culture medium components include mesenchymal stem cell culture medium components and hematopoietic stem cell culture medium components, and the cell culture medium characteristic values ​​include mesenchymal stem cell culture medium characteristic values ​​and hematopoietic stem cell culture medium characteristic values; Determining a cell culture detection reference evaluation value based on a cell culture environment characteristic value and a cell culture medium component characteristic value, wherein the cell culture detection reference evaluation value includes a mesenchymal stem cell culture detection reference evaluation value and a hematopoietic stem cell culture detection reference evaluation value; Test the culture status of mesenchymal stem cells and determine whether the mesenchymal stem cell culture is qualified by combining the mesenchymal stem cell culture test reference evaluation value; Test the hematopoietic stem cell culture status and determine whether the hematopoietic stem cell culture is qualified by combining the hematopoietic stem cell culture test reference evaluation value; Analyzing the unqualified mesenchymal stem cells and hematopoietic stem cells to obtain cell characteristic values, wherein the cell characteristic values ​​include mesenchymal stem cell characteristic values ​​and hematopoietic stem cell characteristic values; Based on the characteristic values ​​of mesenchymal stem cells and hematopoietic stem cells, determine the adjustment plan for the culture of unqualified mesenchymal stem cells and hematopoietic stem cells; The cell culture environment is analyzed, and the specific analysis process is as follows: Acquire a cell culture environment data set, wherein the cell culture environment data set specifically includes cell culture environment temperature, cell culture environment carbon dioxide concentration, and cell culture environment humidity; Based on the acquired cell culture environment data set, a comprehensive analysis is performed to obtain the cell culture environment characteristic value, which is used as the analysis basis for determining the reference evaluation value of the cell culture detection; The cell culture medium components are analyzed to obtain the cell culture medium component characteristic values, and the specific analysis process is as follows: The components of mesenchymal stem cell culture medium were analyzed to obtain the characteristic values ​​of mesenchymal stem cell culture medium; A mesenchymal stem cell culture medium component data set is obtained, wherein the mesenchymal stem cell culture medium component data set specifically includes fetal bovine serum concentration, penicillin concentration, ampicillin concentration, and basic fibroblast growth factor concentration; Based on the obtained mesenchymal stem cell culture medium component data set, the characteristic value of the mesenchymal stem cell culture medium is obtained through comprehensive analysis. The characteristic value of the mesenchymal stem cell culture medium is used as the analytical basis for determining the reference evaluation value of the mesenchymal stem cell culture detection; The components of hematopoietic stem cell culture medium are analyzed to obtain characteristic values ​​of hematopoietic stem cell culture medium; Acquire a hematopoietic stem cell culture medium component data set, wherein the hematopoietic stem cell culture medium component data set specifically includes human serum albumin concentration, stem cell factor concentration, interleukin concentration, and erythropoietin concentration; Based on the obtained hematopoietic stem cell culture medium component data set, a comprehensive analysis is performed to obtain a hematopoietic stem cell culture medium characteristic value, which is used as an analytical basis for determining a reference evaluation value for hematopoietic stem cell culture detection; The specific analysis process of the cell culture medium component characteristic value is as follows: ； In the formula, is the characteristic value of mesenchymal stem cell culture medium, is the characteristic value of hematopoietic stem cell culture medium, is the fetal bovine serum concentration, is the penicillin concentration, is the concentration of ampicillin, is the concentration of basic fibroblast growth factor, is the concentration of human serum albumin, is the concentration of stem cell factor, is the interleukin concentration, is the erythropoietin concentration, For setting The compensation factor, For setting The compensation factor, For setting The compensation factor, For setting The compensation factor, For setting The compensation factor, For setting The compensation factor, For setting The compensation factor, For setting compensation factor.

2. The in vitro culture and detection method of mesenchymal stem cells and hematopoietic stem cells according to claim 1, characterized in that: The cell culture detection reference evaluation value is determined based on the cell culture environment characteristic value and the cell culture medium component characteristic value. The specific analysis process is as follows: The characteristic values ​​of the cell culture environment and the characteristic values ​​of the mesenchymal stem cell culture medium determine the reference evaluation values ​​for the mesenchymal stem cell culture test; The cell culture environment characteristic value and the mesenchymal stem cell culture medium characteristic value are stored as designated tags, and the designated tags are compared with the mesenchymal stem cell culture detection reference evaluation values ​​corresponding to the designated tags stored in the database to obtain the mesenchymal stem cell culture detection reference evaluation values ​​corresponding to the designated tags; The characteristic values ​​of the cell culture environment and the characteristic values ​​of the hematopoietic stem cell culture medium determine the reference evaluation values ​​for hematopoietic stem cell culture testing; The cell culture environment characteristic value and the hematopoietic stem cell culture medium characteristic value are stored as designated labels, and the designated labels are compared with the hematopoietic stem cell culture detection reference evaluation values ​​corresponding to the designated labels stored in the database to obtain the hematopoietic stem cell culture detection reference evaluation values ​​corresponding to the designated labels.

3. The in vitro culture and detection method of mesenchymal stem cells and hematopoietic stem cells according to claim 1, characterized in that: The specific analysis process of judging whether the mesenchymal stem cell culture is qualified is as follows: Obtaining a mesenchymal stem cell culture status data set, which specifically includes a mesenchymal stem cell proliferation rate, a mesenchymal stem cell apoptosis rate, a mesenchymal stem cell differentiation efficiency, and a proportion of mesenchymal stem cell morphological abnormalities; Based on the acquired mesenchymal stem cell culture status data set, a comprehensive analysis is performed to obtain a mesenchymal stem cell culture status evaluation value, which is used as an analysis basis for judging whether the mesenchymal stem cell culture is qualified; comparing the mesenchymal stem cell culture status assessment value with a mesenchymal stem cell culture detection reference assessment value; If the mesenchymal stem cell culture status assessment value is not lower than the mesenchymal stem cell culture detection reference assessment value, the mesenchymal stem cell culture corresponding to the mesenchymal stem cell culture status assessment value is qualified; If the mesenchymal stem cell culture status assessment value is lower than the mesenchymal stem cell culture detection reference assessment value, the mesenchymal stem cell culture corresponding to the mesenchymal stem cell culture status assessment value is unqualified.

4. The in vitro culture and detection method of mesenchymal stem cells and hematopoietic stem cells according to claim 1, characterized in that: The specific analysis process of judging whether the hematopoietic stem cell culture is qualified is as follows: Acquire a hematopoietic stem cell culture status data set, which specifically includes a hematopoietic stem cell proliferation rate, a hematopoietic stem cell apoptosis rate, a hematopoietic stem cell differentiation efficiency, and a proportion of hematopoietic stem cell morphological abnormalities; Based on the acquired hematopoietic stem cell culture status data set, a hematopoietic stem cell culture status evaluation value is obtained through comprehensive analysis, and the hematopoietic stem cell culture status evaluation value is used as an analysis basis for judging whether the hematopoietic stem cell culture is qualified; comparing the hematopoietic stem cell culture status assessment value with a hematopoietic stem cell culture assay reference assessment value; If the hematopoietic stem cell culture status assessment value is not lower than the hematopoietic stem cell culture detection reference assessment value, the hematopoietic stem cell culture corresponding to the hematopoietic stem cell culture status assessment value is qualified; If the hematopoietic stem cell culture status assessment value is lower than the hematopoietic stem cell culture detection reference assessment value, the hematopoietic stem cell culture corresponding to the hematopoietic stem cell culture status assessment value is unqualified.

5. The in vitro culture and detection method of mesenchymal stem cells and hematopoietic stem cells according to claim 1, characterized in that: The analysis of the unqualified mesenchymal stem cells and hematopoietic stem cells is carried out as follows: The characteristic values ​​of mesenchymal stem cells are obtained by analyzing mesenchymal stem cells that fail to meet the culture standards; Obtaining a data set of unqualified mesenchymal stem cells for culture, which specifically includes the number of mesenchymal stem cell passages, the microbial contamination area of ​​the mesenchymal stem cell culture medium, and the bacterial contamination area of ​​the mesenchymal stem cell culture medium; Based on the acquired data set of unqualified mesenchymal stem cells, a comprehensive analysis is performed to obtain the characteristic values ​​of mesenchymal stem cells, which are used as the analysis basis for determining the adjustment plan for unqualified mesenchymal stem cell culture; The characteristic values ​​of hematopoietic stem cells are obtained by analyzing hematopoietic stem cells that fail to meet the culture standards; Acquire a data set of unqualified hematopoietic stem cell culture, which specifically includes the number of hematopoietic stem cell passages, the microbial contamination area of ​​the hematopoietic stem cell culture medium, and the bacterial contamination area of ​​the hematopoietic stem cell culture medium; Based on the acquired data set of unqualified hematopoietic stem cell cultures, a comprehensive analysis is performed to obtain the hematopoietic stem cell characteristic values, which serve as the analytical basis for determining the adjustment plan for unqualified hematopoietic stem cell culture.

6. The in vitro culture and detection method of mesenchymal stem cells and hematopoietic stem cells according to claim 1, characterized in that: The specific analysis process of determining the unqualified mesenchymal stem cell and hematopoietic stem cell culture adjustment plan is as follows: Comparing the mesenchymal stem cell characteristic value with the unqualified mesenchymal stem cell culture adjustment plan corresponding to each mesenchymal stem cell characteristic value stored in the database, to obtain the unqualified mesenchymal stem cell culture adjustment plan corresponding to the mesenchymal stem cell characteristic value; The hematopoietic stem cell characteristic value is compared with the unqualified hematopoietic stem cell culture adjustment plan corresponding to each hematopoietic stem cell characteristic value stored in the database to obtain the unqualified hematopoietic stem cell culture adjustment plan corresponding to the hematopoietic stem cell characteristic value.

7. An in vitro culture detection system for mesenchymal stem cells and hematopoietic stem cells, applied to an in vitro culture detection method for mesenchymal stem cells and hematopoietic stem cells according to any one of claims 1 to 6, characterized in that: It includes an environmental characteristic value acquisition module, a culture medium component characteristic value acquisition module, a culture detection reference evaluation value determination module, a mesenchymal stem cell culture qualification judgment module, a hematopoietic stem cell culture qualification judgment module, a cell characteristic value acquisition module and a culture adjustment plan determination module, wherein: The environment characteristic value acquisition module is used to analyze the cell culture environment to obtain the cell culture environment characteristic value; The culture medium component characteristic value acquisition module is used to analyze the cell culture medium components to obtain the cell culture medium component characteristic values, wherein the cell culture medium components include mesenchymal stem cell culture medium components and hematopoietic stem cell culture medium components, and the cell culture medium characteristic values ​​include mesenchymal stem cell culture medium characteristic values ​​and hematopoietic stem cell culture medium characteristic values; The culture detection reference evaluation value determination module is used to determine the cell culture detection reference evaluation value based on the cell culture environment characteristic value and the cell culture medium component characteristic value, and the cell culture detection reference evaluation value includes the mesenchymal stem cell culture detection reference evaluation value and the hematopoietic stem cell culture detection reference evaluation value; The mesenchymal stem cell culture qualification judgment module is used to detect the mesenchymal stem cell culture state and judge whether the mesenchymal stem cell culture is qualified in combination with the mesenchymal stem cell culture detection reference evaluation value; The hematopoietic stem cell culture qualification judgment module is used to detect the hematopoietic stem cell culture status and judge whether the hematopoietic stem cell culture is qualified in combination with the hematopoietic stem cell culture detection reference evaluation value; The cell characteristic value acquisition module is used to analyze the unqualified mesenchymal stem cells and hematopoietic stem cells to obtain cell characteristic values, wherein the cell characteristic values ​​include the mesenchymal stem cell characteristic values ​​and the hematopoietic stem cell characteristic values; The culture adjustment scheme determination module is used to determine unqualified mesenchymal stem cell and hematopoietic stem cell culture adjustment schemes based on the characteristic values ​​of the mesenchymal stem cells and the hematopoietic stem cells.

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