Method, system, and apparatus for programmed cell freezing.

Abstract

A method, system and device for programmed cell freezing, which belongs to the field of cell freezing technology, includes a temperature drop rate k and a temperature distribution function T=a×h of an insulation cavity in which liquid nitrogen is contained. 2 The method includes a step S1 of acquiring +b×h+c, a step S2 of controlling the cell preservation tube to descend in the heat-preserving cavity at a real-time speed of v=(kb) / 2ah, and acquiring the inner and outer temperatures T1 and T2 of the cell preservation tube in real time at the same time and height during the process of the cell preservation tube descending, a step S3 of determining whether the difference Δt between T1 and T2 exceeds a predetermined temperature difference threshold, and if so, proceeding to S4, otherwise continuing S2 and controlling the cell preservation tube until it reaches a position of a predetermined heat-preserving temperature T0, and a step S4 of controlling the cell preservation tube to stop, and continuing S2 when the difference Δt between T1 and T2 is equal to or less than the predetermined temperature difference threshold. The method, system and device can effectively improve the cell freezing effect and the resuscitation rate.

Description

Cross-reference to Related Applications

[0001] This application claims priority to a Chinese patent application with application number 202110584162.4, filed with the Chinese Patent Office on May 27, 2021, and having the invention title "Cell Program Freezing Method, System, Equipment, Medium and Device", the entire content of which is incorporated herein by reference.

Technical Field

[0002] The present invention belongs to the technical field of cell freezing, and particularly relates to a cell program freezing method, system and device.

Background Art

[0003] In recent years, as modern biomedicine has shifted to the era of molecular medicine and personalized diagnosis and treatment, the demand for large-scale and high-quality biological specimens and related information resources has also been increasing rapidly. Many developed countries attach great importance to the protection and development of human genetic resources, and have established various biobanks, population-based biobanks, and virtual biobanks, regarding them as strategic means for developing the core competitiveness in the field of biomedicine. Establishing a biological cell resource bank and storing biological cells (currently mainly sperm cells, embryos or egg cells, or stem cells) is very important for fields such as germplasm resource conservation, breeding, and reproduction.

[0004] Programmed (programmed) cooling, as an effective cooling method, is widely used in the cell freezing and preservation process. Among them, the liquid nitrogen vaporization method is widely used because its device is simple. In this method, the temperature of different gas layers on the liquid nitrogen surface is used to pre-cool the sample (specimen), and after the pre-cooling is completed, the sample is immersed in liquid nitrogen to complete its freezing.

[0005] However, since the surface temperature of liquid nitrogen is not constant, the cooling rate and pre-cooling temperature of the cell sample in the freezing process are not constant, resulting in a large variation in the preservation effect of the cells.

Summary of the Invention

[0006] In consideration of the problem in the prior art where the cooling rate and pre-cooling temperature do not match during programmed freezing of cells, the present invention aims to provide a programmed freezing method, system, and apparatus for cells. [Means for solving the problem]

[0007] The technical solution of the present invention for achieving the above objective is as follows: In a first aspect, the present invention provides a method for programmed cell freezing. The cooling rate k and the temperature distribution function T = a × h of the insulating cavity containing liquid nitrogen are given by: 2 Step S1 is to obtain +b × h + c, where h is the distance from the temperature measurement point to the upper edge of the heat retention cavity, T is the temperature at the temperature measurement point, and a, b, and c are all constants. Step S2 involves controlling the cell storage tube to descend within the heat-insulating cavity at a real-time speed of v = (kb) / 2ah, and acquiring in real-time the internal and external temperatures T1 and T2 of the cell storage tube at the same time and height during the descent process. Step S3 involves determining whether the difference Δt between the internal temperature T1 and the external temperature T2 exceeds a predetermined temperature difference threshold. If it does, the process proceeds to step S4; otherwise, the cell storage tube is controlled to continue step S2 until it reaches a predetermined temperature retention temperature T0. The method includes step S4, which controls the cell storage tube to stop, and continues step S2 when the difference Δt between the internal temperature T1 and the external temperature T2 falls below the temperature difference threshold.

[0008] Furthermore, step S2, which obtains the internal temperature T1, The steps include: positioning a control tube at the same height as the cell storage tube; The steps include: placing a control solution in the control tube whose thermophysical properties match those of the liquid in the cell preservation tube, or placing the same liquid in the control tube as in the cell preservation tube; The step may include collecting the temperature of the liquid inside the control tube via a temperature-sensing element and determining it as the internal temperature T1.

[0009] Furthermore, in step S3, the position h0 of the predetermined heat retention temperature T0 is determined by the temperature distribution function T = a × h of the heat retention cavity. 2 It may also be calculated using +b × h + c.

[0010] Furthermore, in step S3, controlling the cell storage tube to reach the predetermined warming temperature T0 is This may be achieved by comparing the internal temperature T1 acquired in real time with the predetermined heat retention temperature T0, determining whether the internal temperature T1 is less than the predetermined heat retention temperature T0, controlling the cell storage tube to continue descending if the internal temperature T1 is less than the predetermined heat retention temperature T0, and stopping the cell storage tube if it is not.

[0011] Furthermore, in step S3, after controlling the cell storage tube to reach the predetermined temperature T0, Step S5 may include controlling the cell storage tube so that it rises within the insulated cavity after waiting for a predetermined insulated time, until it is removed from the insulated cavity.

[0012] In a second aspect, the present invention proposes a cell programmed freezing system, the system being used to carry out the above method, the system is An acquisition module used to obtain the cooling rate k, the temperature distribution function of the insulation cavity, T1 and T2 which are the internal and external temperatures of the cell storage tube at the same time and height, a predetermined insulation temperature T0, a predetermined temperature difference threshold, and a predetermined insulation time length. A determination module used to determine whether the difference Δt between the internal temperature T1 and the external temperature T2 exceeds a predetermined temperature difference threshold, and whether the cell storage tube has reached a predetermined temperature T0, The system includes a lifting control module for generating lifting commands according to the judgment result of the judgment module, and for controlling the lifting device to raise and lower the cell storage tube within the heat-retaining cavity according to the lifting commands.

[0013] In a third aspect, the present invention proposes a cell program freezing device. The base and A thermal insulation cavity is placed on the aforementioned base and contains liquid nitrogen inside, A tube rack where cell storage tubes are placed, At least two temperature-measuring elements are installed to measure the temperature at the same height on both the inside and outside of the cell storage tube at the same time, respectively. A lifting device is installed on the base and has an output terminal fixedly connected to the tube rack in order to raise and lower the tube rack within the heat-insulating cavity, The electronic device includes an electronic device, and the electronic device includes a control device that is electrically connected to the temperature measuring element and the lifting device. The electronic device includes a memory for storing executable program code and a processor coupled to the memory, wherein the processor calls the executable program code stored in the memory and performs the method described above.

[0014] Furthermore, the system may include a control tube, the control tube being fixedly installed in the tube rack, and the control tube and the cell storage tube being positioned at the same height. The thermometer may be placed inside the control tube to indirectly obtain the internal temperature of the cell storage tube. [Effects of the Invention]

[0015] According to certain embodiments provided by the present invention, the present invention discloses the following technical effects. The present invention proposes a cell program freezing method, system and device. In the process of controlling the descent of a cell preservation tube in a heat preservation cavity containing liquid nitrogen, by continuously acquiring the inner temperature and outer temperature of the cell preservation tube, during the descent process, it is ensured that the temperature difference between the inside and outside of the cell preservation tube is always lower than a predetermined temperature difference threshold. When the temperature difference exceeds the temperature difference threshold, the descent is stopped, and when the temperature difference is below the temperature difference threshold, the descent is continued until reaching the position of a predetermined heat preservation temperature. Thus, the preservation effect of cells during the programmed freezing process can be effectively improved, and the problem that the preservation effect of cells caused by the inconsistency between the cooling rate and the pre-cooling temperature during the cell program freezing process in the prior art is poor can be solved.

Brief Description of the Drawings

[0016] To more clearly explain the technical solution of the present invention, the drawings necessary in the embodiments or the description of the prior art are briefly introduced below. Obviously, the drawings in the following description are only a part of the embodiments of the present invention, and other drawings can be obtained from these drawings by those skilled in the art without creative labor. [Figure 1] It is a schematic structural diagram of a cell program freezing device provided in an embodiment of the present invention. [Figure 2] It is a flowchart of a cell program freezing method provided in an embodiment of the present invention. [Figure 3] It is a flowchart of a method for acquiring the inner temperature of a cell preservation tube in the cell program freezing method provided in an embodiment of the present invention. [Figure 4] It is a flowchart of another cell program freezing method provided in an embodiment of the present invention. [Figure 5] It is a schematic structural diagram of a cell program freezing system provided in an embodiment of the present invention.

Modes for Carrying Out the Invention

[0017] Specific embodiments of the present invention will be further described below with reference to the drawings. It should be noted that these descriptions of embodiments are provided to aid in understanding the present invention and do not limit it. Furthermore, the technical features of the various embodiments of the present invention described below can be combined with each other, insofar as they do not conflict with each other.

[0018] Furthermore, in the description of this invention, the directions and positional relationships indicated by terms such as "up," "down," "left," "right," "front," and "back" are for the purpose of describing the structure of this invention based on the drawings and are intended solely to provide a brief explanation of the invention. They do not suggest or imply that the devices or elements mentioned must have a specific orientation, or that they must be constructed and operated in a specific orientation, and therefore cannot be interpreted as limitations of this invention.

[0019] In this technical solution, "First" and "Second" are merely distinguishing names for the same or similar structures, or corresponding structures that perform similar functions, and do not represent a ranking of importance, size ranking, comparison, or any other meaning.

[0020] Furthermore, unless explicitly defined and limited, the terms “installation” and “connection” should be understood in a broad sense. For example, a connection may be a fixed connection, a removable connection, or an integrated connection; it may be a mechanical connection or an electrical connection; it may be a direct connection, an indirect connection via an intermediate medium, or internal communication between two structures. Those skilled in the art will understand the specific meaning of the above terms in this invention in conjunction with the specific circumstances of the context of this solution, based on the overall concept of the invention.

[0021] Example 1 The cell program freezing apparatus proposed in this embodiment includes a control device 1, a heat-insulating cavity 3, a tube rack 4, cell storage tubes 6, a temperature sensing element, a lifting device, and a base 14, as shown in Figure 1.

[0022] Here, the base 14 is configured as a platform and is placed on a desk when in use. The thermal cavity 3 is configured as a cylindrical structure with an opening at the top, made of a material with low thermal conductivity (such as foam material), or the thermal cavity 3 can be configured as a vacuum cup. Liquid nitrogen is contained inside the thermal cavity 3, and the liquid nitrogen level is usually at a certain distance, such as 10 cm, from the upper edge of the thermal cavity 3. Furthermore, the temperature distribution above the liquid level in the thermal cavity 3 is given by the temperature distribution function T = a × h 2 The equation +b × h + c is satisfied, where h is the distance from the temperature measurement point to the upper edge of the heat-insulating cavity 3, T is the temperature at the temperature measurement point, and a, b, and c are all constants that can be determined experimentally.

[0023] Here, the cell storage tube 6 is used to hold cells to be frozen, and the cell storage tube 6 is fixedly installed on the tube rack 4. The cell storage tube 6 is driven by the tube rack 4 to move up and down within the insulated cavity 3 and is programmed to cool down. On the other hand, the lifting device is installed on the base 14, and the output end of the lifting device is fixedly connected to the tube rack 4, which drives the tube rack 4 to move up and down within the insulated cavity 3. In this embodiment, the lifting device is configured to include an electric push rod and a curved arm 9. The electric push rod is positioned vertically on the base 14, one end of the curved arm 9 is fixed to the output end of the electric push rod, and the other end of the curved arm 9 is fixedly connected to the tube rack 4. As shown in Figure 1, the electric push rod includes a slider 10, a screw 11, a guide rail 12, and a motor 13. Here, the guide rail 12 is positioned and fixed vertically on the base 14. The screw 11 is parallel to the guide rail 12, and its upper and lower ends are rotatably connected via bearings to the cross plate at the top of the guide rail 12 and the base 14, respectively. The motor 13 is fixed to the base 14, and the output shaft of the motor 13 is connected to the lower end of the screw 11 via a coupling or gear transmission mechanism. The slider 10 is slidably connected to the guide rail 12, and the slider 10 is provided with a screw hole that fits the screw 11, so that when the motor 13 is started, the screw 11 is rotationally driven, and the screw 11 moves the slider 10 up and down, driving the tube rack 4 and the cell storage tubes 6 placed thereon to move up and down within the insulation cavity 3 via the curved arm 9. Alternatively, in another embodiment, the lifting device may be configured as a cylinder, the body of which is fixedly mounted on the base 14, and its output end is used to fix the curved arm 9.

[0024] Furthermore, the thermometer measures the temperature both inside and outside the cell storage tube 6, respectively. For example, in this embodiment, the thermometer is configured to include a first thermometer 7 and a second thermometer 8. Both the first thermometer 7 and the second thermometer 8 are selected as thermocouples, where the probe of the first thermometer 7 extends into the cell storage tube 6 to acquire the internal temperature, and the probe of the second thermometer 8 is located outside the cell storage tube 6 to acquire the external temperature, and the probes of the first thermometer 7 and the second thermometer 8 are at the same height. As will be readily apparent to those skilled in the art, at least two thermometers are arranged to simultaneously acquire the internal and external temperatures of the cell storage tube 6, respectively, but multiple thermometers may be arranged to reduce measurement errors at a single point on one side in order to improve temperature measurement accuracy.

[0025] The control device 1 includes electronic equipment, which includes a memory for storing executable program code and a processor coupled to the memory, and the processor is also electrically connected to the temperature sensing element and the lifting device. The processor calls the executable program code stored in the memory and thereby executes the steps of the cell program freezing method set out in the executable program code.

[0026] When using it, first, depending on the structure of the heat-retaining cavity 3, the temperature distribution function T = a × h is determined experimentally. 2The formula +b×h+c is obtained in advance, and the cells to be frozen are placed in the cell storage tube 6 along with the necessary cell protection agent. Based on the required cooling rate k, the real-time descent rate v=(kb) / 2ah of the cell storage tube 6 is obtained, where k is the cooling rate, h is the distance from the temperature measurement point to the upper edge of the insulation cavity 3, and a, b, and c are all constants. Next, under the control of the control device 1, the lifting device is driven to lower the tube rack 4 and the cell storage tube 6 fixedly installed on it at the specified speed, where h can be calculated from the rotation angle of the motor 13. Next, during the descent process, the temperature difference between the first temperature measuring element 7 and the second temperature measuring element 8 is compared, and it is ensured that the value of the difference is smaller than a predetermined temperature difference threshold. When the value of the difference reaches the temperature difference threshold, the control device 1 stops the descent of the cell storage tube 6. This process is continued until the temperature data measured by the first temperature measuring element 7 reaches the freezing temperature. The apparatus provided in this embodiment can monitor the cell programmed freezing process in real time and make rapid adjustments based on the monitoring results, thereby improving the freezing effect.

[0027] In a preferred embodiment, the control device 1 is further configured to include a display screen 2. The display screen 2 is electrically connected to the processor in the control device 1, and preferably, the display screen 2 is a touchscreen. Control parameters (such as the cooling rate k and the temperature difference threshold) are input and modified via the display screen 2.

[0028] Example 2 The difference between Example 2 and Example 1 is as follows. In Example 2, as shown in Figure 1, a control tube 5 is further included, which is fixedly installed in the tube rack 4, and the control tube 5 and the cell storage tube 6 are positioned at the same height. When in use, the control tube 5 is filled with an equal volume of a control solution whose thermophysical properties (including crystallization enthalpy, thermal conductivity, specific heat capacity, etc.) are the same as those of the liquid in the cell storage tube 6, or an equal volume of the same solution as the cell storage tube 6 is directly filled into the control tube 5. In addition, a thermometer (i.e., a first thermometer 7) is placed inside the control tube 5, and the temperature measured by this first thermometer 7 is taken as the temperature inside the cell storage tube 6 by an indirect measurement method, and at the same time, the first thermometer 7 that was placed inside the cell storage tube 6 in Example 1 is eliminated.

[0029] By installing the device in this manner, it is possible to avoid affecting the cells being frozen by temperature measurement, and further improve the freezing effect of the cells.

[0030] Example 3 This embodiment proposes a cell programmed freezing method, which is carried out based on the apparatus of Example 1, and is performed by the electronic equipment of the apparatus to realize the steps shown in Figure 2. The specific steps include: The cooling rate k and the temperature distribution function T = a × h of the insulating cavity 3 containing liquid nitrogen. 2 We obtain +b × h + c, where h is the distance from the temperature measurement point to the upper edge of the heat-insulating cavity 3, T is the temperature at the temperature measurement point, and a, b, and c are all constants. Step S1, Step S2 involves controlling the cell storage tube 6 to descend within the insulated cavity 3 at a real-time speed of v=(kb) / 2ah via a lifting device, and acquiring in real-time the internal and external temperatures T1 and T2 of the cell storage tube 6 at the same time and height during the descent process. Step S3 involves determining whether the difference Δt (absolute value) between T1 and T2 exceeds a predetermined temperature difference threshold, and if it does, proceeding to S4; otherwise, continuing S2 until the cell storage tube 6 reaches a predetermined temperature T0. Here, the position h0 at a predetermined heat retention temperature T0 is given by the temperature distribution function T = a × h of the heat retention cavity 3. 2 It can be calculated using +b × h + c, or it can be achieved by the following steps: The system compares T1 and T0 values ​​acquired in real time to determine whether T1 is smaller than T0. If T1 is smaller, the lifting device continues to lower the cell storage tube 6; otherwise, the lifting device stops the cell storage tube 6. The above step S3, The process includes step S4, which controls the cell storage tube 6 to stop using a lifting device, and continues S2 when the difference Δt (absolute value) between T1 and T2 falls below a predetermined temperature difference threshold.

[0031] For example, when applying the method provided in this embodiment to sperm freezing, the following occurs: 1. Collect 2 ml of healthy sperm from each of 3 volunteers and allow it to liquefy naturally in a 37°C incubator. 2. Next, using a sperm counting board, first detect sperm motility under a microscope. Repeat the detection twice for each sample, counting 200 sperm each time. Compare the two detection values ​​until the error rate between the two values ​​is within an acceptable range. If the error rate between the two values ​​is not within an acceptable range, re-prepare the sample and re-detect. 3. Sperm are graded as follows: Grade A: moves quickly, Grade B: moves slowly, Grade C: shakes in place, Grade D: does not move. The sperm freezing revival rate (hereinafter referred to as the sperm freezing revival rate) is calculated as follows: Sperm freezing revival rate = (Percentage of Grade A sperm after freezing + Percentage of Grade B sperm after freezing) / (Percentage of Grade A sperm before freezing + Percentage of Grade B sperm before freezing). 4. Mix with commercially available sperm protection solution in a 1:1 ratio, and freeze each mixture in cell preservation tube 6. 5. Install the first temperature measuring element 7 and the second temperature measuring element 8 and freeze according to the above steps. Specifically, the program involves lowering the temperature to -80°C at a cooling rate of 10K / min and holding it for 20 minutes to complete the pre-freezing. Then, it is lowered at 10K / min and frozen in liquid nitrogen at -196°C for 2 weeks before being removed.

[0032] Furthermore, a control group was established and frozen using artificial liquid nitrogen fumigation. Specifically, a cryopreservation tube containing sperm and protective solution was placed 10 cm away from the surface of liquid nitrogen for 10 minutes, and then the cryopreservation tube was immersed in liquid nitrogen for 2 weeks before being removed.

[0033] All frozen sperm were thawed in 37°C warm water, and after thawing, the sperm freezing revival rate was recalculated. The results are shown in Table 1. It can be seen that the revival rate of cells frozen using the method of this example is higher than that of cells frozen using the artificial liquid nitrogen fumigation method.

[0034] [Table 1]

[0035] Example 4 The difference between Example 4 and Example 3 is as follows. In Example 4, in step S2, the T1 temperature inside the cell storage tube 6 is not directly obtained, but indirectly through the following steps. Specifically, as shown in Figure 3, the following steps are included. Step S21: Place the control tube 5 at the same height as the cell storage tube 6. Step S22: Place a control solution in control tube 5 that has the same thermal properties as the liquid in cell storage tube 6, or place the same liquid in control tube 5 as in cell storage tube 6, for example, preferably by adding an equal volume of artificial seminal plasma mixture to control tube 5. Step S23: The temperature of the liquid in the control tube 5 is collected via a thermometer, and this becomes the T1 temperature inside the cell storage tube 6.

[0036] Example 5 The difference between Example 5 and Example 3 is as follows. In Example 5, after step S3 the cell storage tube 6 reaches a predetermined temperature T0, the process further includes step S5, specifically as shown in Figure 4. Step S5: After waiting for a predetermined incubation period, the cell storage tube 6 is controlled by a lifting device to rise within the incubation cavity 3 until it is removed from the incubation cavity 3.

[0037] Example 6 This embodiment proposes a cell programmed freezing system, which is used to carry out the methods of each of the above embodiments. As shown in Figure 5, the system includes: An acquisition module used to obtain the cooling rate k, the temperature distribution function of the insulation cavity 3, the internal temperature T1 and external temperature T2 of the cell storage tube 6 at the same time and height, a predetermined insulation temperature T0, a predetermined temperature difference threshold, and a predetermined insulation time length. It is used to determine whether the difference Δt between T1 and T2 exceeds a predetermined temperature difference threshold, and whether the cell storage tube 6 has reached a predetermined incubation temperature T0. It is easily understood that the difference Δt in each of the above embodiments is an absolute value, and the determination module is used to determine whether the difference Δt exceeds a predetermined temperature difference threshold, The system includes a lifting control module that generates lifting commands according to the judgment result of a judgment module, and is used to control the lifting device so that it lifts and lowers the cell storage tube 6 within the heat retention cavity 3 according to the lifting commands.

[0038] The present invention will be described with reference to flowcharts and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of the present invention. It should be understood that each step and / or block in a flowchart and / or block diagram, as well as combinations of steps and / or blocks in a flowchart and / or block diagram, can be implemented by computer program directives. These computer program directives are provided to the processor of a general-purpose computer, a dedicated computer, an embedded processor, or other programmable data processing device to generate a machine, and the directives executed by the processor of the computer or other programmable data processing device can generate an apparatus to implement one or more steps in a flowchart and / or one or more blocks in a block diagram.

[0039] These computer program instructions may be stored in computer-readable memory that can operate a computer or other programmable data processing device in a specific manner. The instructions stored in computer-readable memory then generate a product including a instruction device. This instruction device implements one or more steps in a flowchart and / or one or more blocks in a block diagram.

[0040] These computer program instructions can be loaded into a computer or other programmable data processing device to execute a series of operational steps on the computer or other programmable device to generate a computer implementation process. Thus, the instructions executed on the computer or other programmable device provide steps to realize one or more steps in a flowchart and / or one or more blocks in a block diagram.

[0041] Although embodiments of the present invention have been described in detail above with reference to the drawings, the present invention is not limited to these embodiments. Those skilled in the art can make various changes, modifications, substitutions, and variations of these embodiments without departing from the principles and spirit of the present invention, and these are also within the scope of protection of the present invention. [Explanation of symbols]

[0042] 1. Control device, 2. Display screen, 3. Insulated cavity, 4. Tube rack, 5. Control tube, 6. Cell storage tube, 7. First temperature sensing element, 8. Second temperature sensing element, 9. Curved arm, 10. Slider, 11. Screw, 12. Guide rail, 13. Motor, 14. Base.

Claims

1. A method for freezing a cell program, The aforementioned cell program freezing method is The cooling rate k and the temperature distribution function T = a × h of the insulated cavity containing liquid nitrogen. ２ Step S1 is to obtain +b × h + c, where h is the distance from the temperature measurement point to the upper edge of the heat-insulating cavity, T is the temperature at the temperature measurement point, and a, b, and c are determined experimentally based on the heat-insulating cavity, and the above step S1 is performed. The cell storage tube is controlled to descend within the heat-insulating cavity at a real-time rate of v = (k - b) / 2ah, and during the process of the cell storage tube descending, the internal and external temperatures of the cell storage tube at the same time and height are T １ and T ２ Step S2 acquires the data in real time, The inside temperature T １ and the outside temperature T ２ The system determines whether the difference Δt exceeds a predetermined temperature difference threshold. If it does, the system proceeds to step S4. Otherwise, the cell storage tube is controlled to maintain a predetermined temperature T. ０ Step S3 is to continue step S2 until the position is reached, Control the cell storage tube to stop, and the internal temperature T １ and the outside temperature T ２ Step S4 includes continuing step S2 when the difference Δt is less than or equal to the temperature difference threshold, In the step S2, the inner temperature T １ The step of obtaining is as follows: The steps include: positioning a control tube at the same height as the cell storage tube; The steps include: placing a control solution in the control tube whose thermophysical properties match those of the liquid in the cell preservation tube, or placing the same liquid in the control tube as in the cell preservation tube; The temperature of the liquid inside the control tube is collected via a temperature sensing element, and the internal temperature T is measured. １ This includes the step of, In step S3, the predetermined heat retention temperature T ０ position h ０ The temperature distribution function of the aforementioned heat-retaining cavity is T = a × h ２ It is calculated by +b × h + c, In step S3, the cell storage tube is controlled to maintain the predetermined temperature T ０ Reaching the position is The internal temperature T acquired in real time １ and the predetermined heat retention temperature T ０ , and the inside temperature T １ The predetermined heat retention temperature T ０ Determine whether it is smaller, and the internal temperature T １ The predetermined heat retention temperature T ０ A cell programmed freezing method characterized by being achieved by the step of controlling the cell storage tube to continue descending if it is smaller, and stopping the cell storage tube otherwise.

2. In step S3, the cell storage tube is controlled to maintain a predetermined temperature T ０ After reaching the position, The cell programming freezing method according to claim 1, further comprising step S5 of controlling the cell storage tube so that it rises within the insulated cavity after waiting for a predetermined insulated time, until it is removed from the insulated cavity.

3. A cell program freezing system, the cell program freezing system is used to perform the cell program freezing method described in any one of claims 1 to 2, and the cell program freezing system includes: The cooling rate k, the temperature distribution function of the insulation cavity, and T are the internal and external temperatures of the cell storage tube at the same time and height. １ and T ２ , predetermined heat retention temperature T ０ an acquisition module used to obtain a predetermined temperature difference threshold and a predetermined heat retention time length, The inside temperature T １ and the outside temperature T ２ The system determines whether the difference Δt exceeds a predetermined temperature difference threshold, and whether the cell storage tube is kept at a predetermined temperature T ０ A determination module used to determine whether the position has been reached, A lifting control module is used to generate a lifting command according to the judgment result of the judgment module, and to control the lifting device to lift the cell storage tube up and down within the heat-retaining cavity according to the lifting command, A cell program freezing system characterized by including the following:

4. A cell program freezing apparatus for performing the cell program freezing method according to any one of claims 1 to 2, The base and A thermal insulation cavity is placed on the aforementioned base and contains liquid nitrogen inside, A tube rack where cell storage tubes are placed, At least two temperature-measuring elements are installed to measure the temperature at the same height on both the inside and outside of the cell storage tube at the same time, A lifting device is installed on the base and has an output terminal fixedly connected to the tube rack in order to raise and lower the tube rack within the heat-insulating cavity, The electronic device includes a control device that is electrically connected to the temperature measuring element and the lifting device, A control tube and Includes, The control tube is fixedly installed in the tube rack, and the control tube and the cell storage tube are positioned at the same height. The temperature sensing element is placed inside the control tube to indirectly obtain the temperature inside the cell storage tube. The electronic device includes a memory for storing executable program code and a processor coupled to the memory, wherein the processor calls the executable program code stored in the memory. A cell program freezing device characterized by the following features.

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