A stem cell collection control system

Abstract

The present invention relates to the technical field of stem cell collection, and in particular to a stem cell collection control system, which comprises an image acquisition module, a centrifuge, a magnetic bead sorting column, a flow cytometer and a control mechanism, wherein the control mechanism comprises a data acquisition module and a data analysis module for determining a stem cell collection method according to an activity evaluation value of the stem cells, wherein the data analysis module determines three collection methods according to the activity evaluation value: the first collection method is a density gradient centrifugation method, the second collection method is a magnetic bead sorting method, and the third collection method is a flow cytometric sorting method, and the data analysis module calculates the activity evaluation value according to the total number of stem cells and the number of positive stem cells in a test sample. The present invention overcomes the problems of poor accuracy in stem cell sorting and great influence on stem cell activity during the sorting process in the prior art.

Description

Technical Field

[0001] The present invention relates to the technical field of stem cell collection, and in particular to a stem cell collection control system. Background Art

[0002] Stem cells are cells that have the ability to self-replicate and differentiate into a variety of cell types. Therefore, they have broad application prospects in the fields of tissue repair, regenerative medicine, and disease treatment. However, the acquisition and collection of stem cells is a technical challenge. Existing stem cell collection methods mainly include density gradient centrifugation, magnetic bead sorting, and flow cytometry sorting, but these methods have their own advantages and disadvantages. How to choose the most appropriate collection method to improve the efficiency and quality of stem cell collection is a major challenge.

[0003] Density gradient centrifugation is a commonly used cell sorting method. Its advantage is that it is easy to operate, but the sorting efficiency is low, and multiple sortings may be required to obtain the desired stem cells. Magnetic bead sorting uses a magnetic field to sort out cells with specific markers. Its sorting efficiency is high, but it may affect the activity of stem cells. Flow cytometry sorting can obtain the desired stem cells in one sorting, and can perform high-precision sorting based on the physical and chemical properties of stem cells, but the equipment is complex and the operation is difficult.

[0004] Chinese Patent Publication No.: CN111004707A discloses a stem cell collection control system and a control method thereof, including a controller, an actuator, a peristaltic pump assembly and a sensor assembly, wherein the actuator, the peristaltic pump assembly and the sensor assembly are respectively connected to the controller, wherein the controller is used to output a control signal; the actuator is used to work according to the control signal to drive the peristaltic pump assembly to a specified position; the peristaltic pump assembly is used to absorb cells according to the control signal; the sensor assembly is used to detect whether the actuator has reached the specified position and whether the peristaltic pump assembly has absorbed all the cells, and to form an input signal to input into the controller; the sensor assembly also includes a photoelectric sensor for detecting whether the actuator has reached the specified position, a position sensor and an ultrasonic sensor for detecting the liquid level where the cells are located.

[0005] It can be seen that the existing technology has the following problems: the existing stem cell collection method has an impact on the activity of stem cells, and the accuracy of stem cell sorting is poor, and it is unable to cope with the task of sorting stem cells at different activity levels. Summary of the invention

[0006] To this end, the present invention provides a stem cell collection control system to overcome the problems in the prior art of poor accuracy in stem cell sorting and significant impact on stem cell activity during the sorting process.

[0007] To achieve the above objectives, in one aspect, the present invention provides a stem cell collection control system, comprising:

[0008] An image acquisition module, used for acquiring images of stem cells in the detection sample;

[0009] A centrifuge for performing density gradient centrifugation on a sample to separate stem cells from the sample;

[0010] A magnetic bead sorting column, used to sort out stem cells in the sample by magnetic bead sorting and flow them into a beaker, comprising a coil wound around the outside of the sorting column for controlling the magnetic field strength and magnetic beads arranged inside the sorting column for sorting stem cells;

[0011] A flow cytometer, used to perform flow cell sorting on a sample to sort out stem cells in the sample;

[0012] A sample storage box, which is connected to the image acquisition module, the centrifuge, the magnetic bead separation column and the flow cytometer, and is used to store stem cell samples and transport the stem cell samples to the image acquisition module, the centrifuge, the magnetic bead separation column and the flow cytometer;

[0013] A stem cell storage box, which is connected to the centrifuge, beaker and flow cytometer, and is used to store the sorted stem cells;

[0014] The control mechanism includes a data acquisition module for acquiring the image of stem cells in the test sample acquired by the image acquisition module and a data analysis module for analyzing the data acquired by the data acquisition module and controlling the centrifuge, the magnetic bead separation column and the flow cytometer according to the analysis results;

[0015] The data analysis module determines a stem cell collection method according to the activity evaluation value of the stem cells. When the stem cell collection method is determined to be the third collection method, the data analysis module determines several sorting types of the third collection method according to the second relative difference.

[0016] Furthermore, the data analysis module is provided with several collection methods for determining the stem cells according to the comparison result of the activity evaluation value of the stem cells with the preset activity evaluation value, wherein the several collection methods include a first collection method using density gradient centrifugation, a second collection method using magnetic bead sorting method and a third collection method using flow cytometry sorting method.

[0017] Furthermore, the data analysis module calculates the activity evaluation value of stem cells according to the following formula:

[0018] P=B / B0×100%

[0019] Among them, P is the activity evaluation value of stem cells, B0 is the total number of stem cells in the test sample, and B is the number of positive stem cells in the test sample.

[0020] Furthermore, under the first collection method, the data analysis module calculates the coefficient of variation of the volume of stem cells in the test sample and sets

[0021] CV=SD / Mean×100%

[0022] Among them, CV is the coefficient of variation of stem cell volume, SD is the standard deviation of stem cell volume in the test sample, and Mean is the mean value of stem cell volume in the test sample;

[0023] The data analysis module determines the rotation speed of the centrifuge according to the comparison result of the stem cell volume variation coefficient and the preset stem cell volume variation coefficient.

[0024] Furthermore, the data analysis module calculates the relative difference between the activity evaluation value of the stem cells and the first preset activity evaluation value under the second collection method, and determines the magnetic field strength of the coil according to a comparison result between the first relative difference and the first preset relative difference.

[0025] Further, after the stem cells are collected by the second collection method, the data analysis module calculates the sorting rate of the sorted stem cells and sets

[0026] D=(Da×Db) / (DA×DB)

[0027] Wherein, D represents the sorting yield, Da represents the total number of cells in the test sample after sorting, Db represents the percentage of stem cells in the test sample after sorting, DA represents the total number of cells in the test sample before sorting, and DB represents the percentage of stem cells in the test sample before sorting;

[0028] The data analysis module determines to adjust the magnetic field strength according to the sorting yield being less than or equal to the preset sorting yield.

[0029] Furthermore, the data analysis module calculates a first difference between the sorting yield and a preset sorting yield under the condition of determining the adjustment of the magnetic field strength, and determines a number of adjustment coefficients for adjusting the magnetic field strength according to a comparison result of the first difference and the second preset difference, the number of adjustment coefficients including a first adjustment coefficient determined when the first difference is less than or equal to the second preset difference and a second adjustment coefficient determined when the first difference is greater than the second preset difference.

[0030] Furthermore, the data analysis module calculates a second relative difference between the activity evaluation value of the stem cells and a second preset activity evaluation value when determining that the collection method of the stem cells is the third collection method, and determines several sorting types of the flow cytometry sorting method based on the comparison result of the second relative difference and the second preset relative difference, the sorting types including a first sorting type of capture tube sorting and a second sorting type of charge sorting.

[0031] Furthermore, after the stem cells are collected using the third collection method, the data analysis module calculates a third relative difference between the activity of the sorted stem cells and the second preset activity evaluation value, and determines to adjust the second preset activity evaluation value based on the third relative difference being less than the second relative difference.

[0032] Furthermore, when determining to adjust the second preset activity evaluation value, the data analysis module calculates a second difference between the third relative difference and the second relative difference, and determines a number of adjustment coefficients of the second preset activity evaluation value based on a comparison result of the second difference and the second preset difference to adjust the second preset activity evaluation value, wherein the several adjustment coefficients include a first adjustment coefficient determined when the second difference is less than or equal to the second preset difference and a second adjustment coefficient determined when the second difference is greater than the second preset difference.

[0033] Compared with the prior art, the present invention has the beneficial effect that, by determining different collection methods according to the activity evaluation values ​​of stem cells, the present invention improves the flexibility and efficiency of stem cell collection and avoids the loss of stem cell activity due to the use of inappropriate collection methods.

[0034] Furthermore, the present invention determines the rotation speed of the centrifuge according to the coefficient of variation of stem cell volume, thereby more accurately controlling the stem cell collection process, thereby improving the accuracy of stem cell sorting, avoiding stem cell damage caused by excessive rotation speed, and protecting the activity of stem cells.

[0035] Furthermore, the present invention determines the magnetic field strength based on the relative difference between the activity evaluation value of the stem cells and the first preset activity evaluation value, thereby avoiding stem cell damage caused by excessive magnetic field strength, thereby protecting the activity of the stem cells and more accurately controlling the stem cell collection process, thereby improving the accuracy of stem cell collection.

[0036] Furthermore, by comparing the sorting yield with the preset sorting yield, the data analysis module of the present invention dynamically determines whether the magnetic field strength needs to be adjusted, thereby improving the sorting yield by adjusting the magnetic field strength, while avoiding possible damage to stem cells caused by an overly strong magnetic field.

[0037] Furthermore, the present invention calculates a first difference between the sorting yield and a preset sorting yield, and determines an adjustment coefficient of the magnetic field strength according to the first difference, thereby more accurately controlling the magnetic field strength and avoiding stem cell damage caused by excessive magnetic field strength, thereby protecting the activity of the stem cells and improving the stem cell sorting yield.

[0038] Furthermore, the present invention determines the sorting type of flow cytometry sorting by a second relative difference between an activity evaluation value of a stem cell and a second preset activity evaluation value; when the second relative difference is less than or equal to the second preset relative difference, a sorting type with less influence on stem cell activity, namely, capture tube sorting, is selected; and when the second relative difference is greater than the second preset relative difference, a sorting type with a higher sorting yield, namely, charge sorting, is selected, thereby improving the sorting accuracy while ensuring the activity of stem cells.

[0039] Furthermore, when the third relative difference between the activity of the stem cells after sorting of the present invention and the second preset activity evaluation value is less than the second relative difference, it means that the activity of the stem cells after sorting is reduced. At this time, the second preset activity evaluation value needs to be adjusted to select a magnetic bead sorting method that has less impact on the activity of the stem cells, which helps to improve the activity of the stem cells.

[0040] Furthermore, the present invention determines the second preset activity evaluation value adjustment coefficient by comparing the second difference with the second preset difference to more accurately adjust the second preset activity evaluation value, and selects a magnetic bead sorting method that has less impact on stem cell activity, which helps to improve the activity of stem cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 This is a structural diagram of a stem cell collection control system according to an embodiment of the present invention;

[0042] Figure 2 This is a structural diagram of a magnetic bead separation column of a stem cell collection control system according to an embodiment of the present invention;

[0043] Figure 3 This is a schematic diagram of the structure of a stem cell collection control system according to an embodiment of the present invention;

[0044] In the figure, 1-image acquisition module, 2-centrifuge, 3-magnetic bead sorting column, 4-sorting column, 5-coil, 6-magnetic beads, 7-flow cytometer, 8-sample storage box, 9-stem cell storage box, 10-beaker. DETAILED DESCRIPTION

[0045] In order to make the objects and advantages of the present invention more clearly understood, the present invention is further described below in conjunction with embodiments; it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0046] The preferred embodiments of the present invention are described below with reference to the accompanying drawings. It should be understood by those skilled in the art that these embodiments are only used to explain the technical principles of the present invention and are not intended to limit the protection scope of the present invention.

[0047] In addition, it should be noted that in the description of the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0048] See also Figure 1 , Figure 2 and Figure 3 As shown, Figure 1 This is a structural diagram of a stem cell collection control system according to an embodiment of the present invention; Figure 2 This is a structural diagram of a magnetic bead separation column of a stem cell collection control system according to an embodiment of the present invention; Figure 3 Schematic diagram of the structure of a stem cell collection control system according to an embodiment of the present invention.

[0049] The stem cell collection control system of the embodiment of the present invention includes:

[0050] An image acquisition module 1 is used to acquire images of stem cells in the detection sample;

[0051] Centrifuge 2, used for performing density gradient centrifugation on the sample to separate stem cells from the sample;

[0052] A magnetic bead sorting column 3 is used to sort out stem cells in the sample by magnetic bead sorting and flow them into a beaker 10, which includes a coil 5 wound around the outside of the sorting column 4 for controlling the magnetic field strength and magnetic beads 6 arranged inside the sorting column for sorting stem cells;

[0053] A flow cytometer 7, used to perform flow cell sorting on the sample to sort out stem cells in the sample;

[0054] A sample storage box 8, which is connected to the image acquisition module 1, the centrifuge 2, the magnetic bead sorting column 3 and the flow cytometer 7, and is used to store stem cell samples and transport stem cell samples to the image acquisition module 1, the centrifuge 2, the magnetic bead sorting column 3 and the flow cytometer 7;

[0055] The stem cell storage box 9 is connected to the centrifuge 1, the beaker 10 and the flow cytometer 7, and is used to store the sorted stem cells.

[0056] The control mechanism includes a data acquisition module for acquiring the image of stem cells in the test sample acquired by the image acquisition module 1 and a data analysis module for analyzing the data acquired by the data acquisition module and controlling the centrifuge 2, the magnetic bead sorting column 3 and the flow cytometer 7 according to the analysis results.

[0057] In an embodiment of the present invention, the image acquisition module is an optical microscope with an imaging system.

[0058] Specifically, the data analysis module determines the collection method of stem cells according to the activity evaluation value P of the stem cells and a preset activity evaluation value, and the data analysis module is provided with a first preset activity evaluation value P1 and a second preset activity evaluation value P2;

[0059] If P≤P1, the data analysis module determines that the collection method of stem cells is the first collection method;

[0060] If P1<P≤P2, the data analysis module determines that the collection method of stem cells is the second collection method;

[0061] If P>P2, the data analysis module determines that the collection method of stem cells is the third collection method;

[0062] Among them, the first collection method is density gradient centrifugation, the second collection method is magnetic bead sorting, and the third collection method is flow cytometry sorting.

[0063] In the embodiment of the present invention, the first preset activity evaluation value P1 is 0.8, and the second preset activity evaluation value P2 is 0.9. Those skilled in the art may adjust the preset activity evaluation values ​​according to specific circumstances.

[0064] Specifically, the present invention improves the flexibility and efficiency of stem cell collection by determining different collection methods according to the activity evaluation values ​​of stem cells, thereby avoiding the loss of stem cell activity due to the use of inappropriate collection methods.

[0065] Specifically, the data analysis module calculates the activity evaluation value P of stem cells according to the following formula:

[0066] P=B / B0×100%

[0067] Among them, B0 is the total number of stem cells in the test sample, and B is the number of positive stem cells in the test sample.

[0068] In the embodiment of the present invention, BrdU is added to the test sample, and BrdU is synthesized into DNA by actively proliferating stem cells. Samples are taken for observation, and immunohistochemical staining is performed with anti-BrdU antibodies to detect cells labeled with BrdU, i.e., stem cells undergoing DNA replication and proliferation. The number of stem cells undergoing DNA replication and proliferation is recorded as the number of positive stem cells.

[0069] Specifically, when the data analysis module determines that the collection method of stem cells is the first collection method, the data analysis module calculates the coefficient of variation CV of the volume of stem cells in the test sample and sets

[0070] CV=SD / Mean×100%

[0071] Among them, SD is the standard deviation of the stem cell volume in the test sample, and Mean is the mean value of the stem cell volume in the test sample.

[0072] Specifically, when the data analysis module determines that the stem cell collection method is the first collection method, the data analysis module calculates the stem cell volume variation coefficient CV in the sample, and determines the speed of the centrifuge according to the comparison result of the stem cell volume variation coefficient CV and the preset stem cell volume variation coefficient, and the data analysis module is provided with a first preset stem cell volume variation coefficient CV1 and a second preset stem cell volume variation coefficient CV2, wherein CV1<CV2;

[0073] If CV≤CV1, ​​the data analysis module determines that the rotation speed of the centrifuge is a first rotation speed;

[0074] If CV1<CV≤CV2, the data analysis module determines that the rotation speed of the centrifuge is a second rotation speed;

[0075] If CV>CV2, the data analysis module determines that the speed of the centrifuge is a third speed;

[0076] Among them, the first speed is 600 rpm / min, the second speed is 800 rpm / min, and the third speed is 1000 rpm / min.

[0077] In the embodiment of the present invention, the first preset stem cell volume variation coefficient CV1 is 10%, and the second preset stem cell volume variation coefficient CV2 is 15%. Those skilled in the art can adjust the preset stem cell volume variation coefficient according to specific circumstances.

[0078] Specifically, the present invention determines the rotation speed of the centrifuge according to the coefficient of variation of stem cell volume, and more accurately controls the stem cell collection process, thereby improving the accuracy of stem cell sorting, avoiding stem cell damage caused by excessive rotation speed, and protecting the activity of the stem cells.

[0079] Specifically, the data analysis module calculates the relative difference ΔP between the activity evaluation value P of the stem cells and the first preset activity evaluation value P1 under the second collection method, determines the magnetic field intensity Ti according to the comparison result of the first relative difference ΔP1 and the first preset relative difference ΔP10, and sets ΔP1=(P-P1) / P1;

[0080] If △P1≤△P10, the data analysis module determines that the magnetic field strength is the first magnetic field strength T1;

[0081] If ΔP1>ΔP10, the data analysis module determines that the magnetic field strength is the second magnetic field strength T2;

[0082] The first magnetic field strength T1 is 1 Tesla, and the second magnetic field strength T2 is 1.5 Tesla.

[0083] In the embodiment of the present invention, the first relative difference ΔP1 is set to 0.06, and those skilled in the art may adjust the first relative difference according to specific circumstances.

[0084] Specifically, the present invention determines the magnetic field strength based on the relative difference between the activity evaluation value of the stem cells and the first preset activity evaluation value, thereby avoiding stem cell damage caused by excessive magnetic field strength, thereby protecting the activity of the stem cells and more accurately controlling the stem cell collection process, thereby improving the accuracy of stem cell collection.

[0085] Specifically, after the stem cells are collected by the second collection method, the data analysis module calculates the sorting rate D of the sorted stem cells and sets

[0086] D=(Da×Db) / (DA×DB)

[0087] Da represents the total number of cells in the test sample after sorting, Db represents the percentage of stem cells in the test sample after sorting, DA represents the total number of cells in the test sample before sorting, and DB represents the percentage of stem cells in the test sample before sorting.

[0088] Specifically, after the collection of stem cells by the second collection method is completed, the data analysis module determines whether to adjust the magnetic field intensity Ti according to the comparison result between the sorting yield D and the preset sorting yield D0;

[0089] If D≤D0, the data analysis module determines to adjust the magnetic field intensity Ti;

[0090] If D>D0, the data analysis module determines not to adjust the magnetic field intensity Ti;

[0091] In the embodiment of the present invention, the preset sorting yield D0 is set to 0.8, and those skilled in the art can adjust the preset sorting yield according to specific circumstances.

[0092] Specifically, the present invention compares the sorting yield with the preset sorting yield, and the data analysis module dynamically determines whether the magnetic field strength needs to be adjusted. The sorting yield is improved by adjusting the magnetic field strength, while avoiding possible damage to stem cells caused by an overly strong magnetic field.

[0093] Specifically, the data analysis module calculates the first difference △D between the sorting yield D and the preset sorting yield D0 under the condition of determining the adjustment of the magnetic field intensity Ti, and determines the adjustment coefficient yi for adjusting the magnetic field intensity Ti according to the comparison result of the first difference △D and the second preset difference △D0. The data analysis module is provided with a first adjustment coefficient y1 and a second adjustment coefficient y2, and D=D0-D is set;

[0094] If ΔD≤ΔD0, the data analysis module determines to adjust the first magnetic field intensity T1 with the first adjustment coefficient y1;

[0095] If ΔD>ΔD0, the data analysis module determines to adjust the second magnetic field strength T2 with the second adjustment coefficient y2;

[0096] The adjusted first magnetic field strength T10 = T1×yi, i=1,2, and the adjusted second magnetic field strength T20 = T2×yi, i=1,2.

[0097] The first adjustment coefficient y1 is 1.1, and the second adjustment coefficient y2 is 1.2.

[0098] In the embodiment of the present invention, the first preset difference ΔD0 is set to 5%. Those skilled in the art may adjust the first preset difference according to specific circumstances.

[0099] Specifically, the present invention calculates the first difference between the sorting yield and the preset sorting yield, and determines the adjustment coefficient of the magnetic field strength according to the first difference, so as to more accurately control the magnetic field strength and avoid stem cell damage caused by excessive magnetic field strength, thereby protecting the activity of stem cells and improving the stem cell sorting yield.

[0100] Specifically, when the data analysis module determines that the collection method of stem cells is the third collection method, the data analysis module calculates the second relative difference △P2 between the activity evaluation value P of the stem cells and the second preset activity evaluation value P2, and determines the sorting type of the flow cell sorting method according to the comparison result between the second relative difference △P2 and the second preset relative difference △P20, and sets △P2=(P-P2) / ((P+P2)2 / 2);

[0101] If ΔP2≤ΔP20, the data analysis module determines that the sorting type of the flow cell sorting method is the first sorting type;

[0102] If ΔP2>ΔP20, the data analysis module determines that the sorting type of the flow cell sorting method is the second sorting type;

[0103] Among them, the first sorting type is capture tube sorting, and the second sorting type is charge sorting.

[0104] In the embodiment of the present invention, the second relative difference ΔP2 is set to 0.03, and those skilled in the art can adjust the second relative difference according to specific circumstances.

[0105] Specifically, the present invention determines the sorting type of flow cytometry sorting by the second relative difference between the activity evaluation value of the stem cell and the second preset activity evaluation value. When the second relative difference is less than or equal to the second preset relative difference, the sorting type with less impact on the activity of the stem cell, namely, capture tube sorting, is selected. When the second relative difference is greater than the second preset relative difference, the sorting type with a higher sorting yield, namely, charge sorting, is selected, thereby improving the sorting accuracy while ensuring the activity of the stem cells.

[0106] Specifically, after the stem cells are collected by the third collection method, the data analysis module calculates a third relative difference ΔPp between the activity Pp of the sorted stem cells and the second preset activity evaluation value P2, and determines whether to adjust the second preset activity evaluation value P2 according to a comparison result of the third relative difference ΔPp and the second relative difference ΔP2, and sets ΔPp=(Pp-P2) / ((Pp+P2)2 / 2);

[0107] If ΔPp<ΔP2, the data analysis module determines to adjust the second preset activity evaluation value P2;

[0108] If ΔPp≥ΔP2, the data analysis module determines not to adjust the second preset activity evaluation value P2.

[0109] Specifically, when the third relative difference between the activity of the stem cells after sorting of the present invention and the second preset activity evaluation value is less than the second relative difference, it means that the activity of the stem cells after sorting is reduced. At this time, the second preset activity evaluation value needs to be adjusted to select a magnetic bead sorting method that has less impact on the activity of the stem cells, which helps to improve the activity of the stem cells.

[0110] Specifically, when the data analysis module determines to adjust the second preset activity evaluation value P2, the data analysis module calculates the second difference △Pp1 between the third relative difference △Pp and the second relative difference △P2, and determines the adjustment coefficient ki of the second preset activity evaluation value P2 according to the comparison result of the second difference △Pp1 and the second preset difference △Pp0 to adjust the second preset activity evaluation value P2, and the data analysis module is provided with a first adjustment coefficient k1 and a second adjustment coefficient k2, wherein k1<k2, and △Pp1=△P2-△Pp is set;

[0111] If △Pp1≤△Pp0, the data analysis module determines to adjust the second preset activity evaluation value P2 by the first adjustment coefficient k1;

[0112] If ΔPp1>ΔPp0, the data analysis module determines to adjust the second preset activity evaluation value P2 by the second adjustment coefficient k2;

[0113] The adjusted second preset activity evaluation value P02=P2×ki, i=1,2.

[0114] The first adjustment coefficient k1 is set to 1.1, and the second adjustment coefficient k2 is set to 1.2.

[0115] In the embodiment of the present invention, the second preset difference △Pp0 is set to 0.01, and those skilled in the art can adjust the second preset difference according to specific circumstances.

[0116] Specifically, the present invention determines the second preset activity evaluation value adjustment coefficient by comparing the second difference with the second preset difference to more accurately adjust the second preset activity evaluation value, selects a magnetic bead sorting method that has less impact on stem cell activity, and helps to improve the activity of stem cells.

[0117] So far, the technical solutions of the present invention have been described in conjunction with the preferred embodiments shown in the accompanying drawings. However, it is easy for those skilled in the art to understand that the protection scope of the present invention is obviously not limited to these specific embodiments. Without departing from the principle of the present invention, those skilled in the art can make equivalent changes or substitutions to the relevant technical features, and the technical solutions after these changes or substitutions will fall within the protection scope of the present invention.

[0118] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A stem cell collection control system, characterized in that: include: An image acquisition module, used for acquiring images of stem cells in the detection sample; A centrifuge for performing density gradient centrifugation on a sample to separate stem cells from the sample; A magnetic bead sorting column, used to sort out stem cells in the sample by magnetic bead sorting and flow them into a beaker, comprising a coil wound around the outside of the sorting column for controlling the magnetic field strength and magnetic beads arranged inside the sorting column for sorting stem cells; A flow cytometer, used to perform flow cell sorting on a sample to sort out stem cells in the sample; A sample storage box, which is connected to the image acquisition module, the centrifuge, the magnetic bead separation column and the flow cytometer, and is used to store stem cell samples and transport the stem cell samples to the image acquisition module, the centrifuge, the magnetic bead separation column and the flow cytometer; A stem cell storage box, which is connected to the centrifuge, beaker and flow cytometer, and is used to store the sorted stem cells; The control mechanism includes a data acquisition module for acquiring the image of stem cells in the test sample acquired by the image acquisition module and a data analysis module for analyzing the data acquired by the data acquisition module and controlling the centrifuge, the magnetic bead separation column and the flow cytometer according to the analysis results; The data analysis module determines the collection method of stem cells according to the activity evaluation value P of the stem cells and the preset activity evaluation value, and the data analysis module is provided with a first preset activity evaluation value P1 and a second preset activity evaluation value P2; If P≤P1, the data analysis module determines that the collection method of stem cells is the first collection method; If P1<P≤P2, the data analysis module determines that the collection method of stem cells is the second collection method; If P>P2, the data analysis module determines that the collection method of stem cells is the third collection method; Among them, the first collection method is density gradient centrifugation, the second collection method is magnetic bead sorting, and the third collection method is flow cytometry sorting; The first preset activity evaluation value P1 is set to 0.8, and the second preset evaluation value P2 is set to 0.9; The data analysis module calculates the activity evaluation value of stem cells according to the following formula: P=B / B0×100% Wherein, P is the activity evaluation value of stem cells, B0 is the total number of stem cells in the test sample, and B is the number of positive stem cells in the test sample. BrdU is added to the test sample, and BrdU is synthesized into DNA by actively proliferating stem cells. Samples are taken for observation, and immunohistochemical staining is performed with anti-BrdU antibodies to detect cells marked with BrdU, i.e., stem cells that are undergoing DNA replication and proliferation. The number of stem cells undergoing DNA replication and proliferation is recorded as the number of positive stem cells.

2. The stem cell collection control system according to claim 1, characterized in that: The data analysis module calculates the coefficient of variation of the volume of stem cells in the test sample under the first collection method, and sets CV=SD / Mean×100% Among them, CV is the coefficient of variation of stem cell volume, SD is the standard deviation of stem cell volume in the test sample, and Mean is the mean value of stem cell volume in the test sample; The data analysis module calculates the coefficient of variation CV of the stem cell volume in the sample, and determines the speed of the centrifuge according to the comparison result of the coefficient of variation CV of the stem cell volume with the preset coefficient of variation of the stem cell volume, wherein the data analysis module is provided with a first preset coefficient of variation CV1 of the stem cell volume and a second preset coefficient of variation CV2 of the stem cell volume, wherein CV1<CV2; If CV≤CV1, ​​the data analysis module determines that the rotation speed of the centrifuge is a first rotation speed; If CV1<CV≤CV2, the data analysis module determines that the rotation speed of the centrifuge is a second rotation speed; If CV>CV2, the data analysis module determines that the speed of the centrifuge is a third speed; Among them, the first speed is 600rpm, the second speed is 800rpm, the third speed is 1000rpm, the first preset stem cell volume variation coefficient is set to 10%, and the second preset stem cell volume variation coefficient is set to 15%.

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