WO2018148982A1 - 一种细胞冻干系统和方法 - Google Patents

一种细胞冻干系统和方法 Download PDF

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Publication number
WO2018148982A1
WO2018148982A1 PCT/CN2017/074762 CN2017074762W WO2018148982A1 WO 2018148982 A1 WO2018148982 A1 WO 2018148982A1 CN 2017074762 W CN2017074762 W CN 2017074762W WO 2018148982 A1 WO2018148982 A1 WO 2018148982A1
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Prior art keywords
lyophilization
work box
cell
nitrogen
cell sample
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PCT/CN2017/074762
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English (en)
French (fr)
Inventor
徐小杨
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徐小杨
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Application filed by 徐小杨 filed Critical 徐小杨
Priority to JP2019565599A priority Critical patent/JP6883672B2/ja
Priority to RU2019128542A priority patent/RU2734536C1/ru
Priority to EP17896492.0A priority patent/EP3583847B1/en
Priority to US16/487,154 priority patent/US11399537B2/en
Publication of WO2018148982A1 publication Critical patent/WO2018148982A1/zh

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    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F26—DRYING
    • F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
    • F26B5/00—Drying solid materials or objects by processes not involving the application of heat
    • F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
    • F26B5/06—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum the process involving freezing
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0278—Physical preservation processes
    • A01N1/0284—Temperature processes, i.e. using a designated change in temperature over time
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0236—Mechanical aspects
    • A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
    • A01N1/0252—Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
    • A01N1/0257—Stationary or portable vessels generating cryogenic temperatures
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0236—Mechanical aspects
    • A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
    • A01N1/0268—Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F26—DRYING
    • F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
    • F26B21/00—Arrangements or duct systems, e.g. in combination with pallet boxes, for supplying and controlling air or gases for drying solid materials or objects
    • F26B21/14—Arrangements or duct systems, e.g. in combination with pallet boxes, for supplying and controlling air or gases for drying solid materials or objects using gases or vapours other than air or steam, e.g. inert gases
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F26—DRYING
    • F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
    • F26B23/00—Heating arrangements
    • F26B23/10—Heating arrangements using tubes or passages containing heated fluids, e.g. acting as radiative elements; Closed-loop systems
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F26—DRYING
    • F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
    • F26B25/00—Details of general application not covered by group F26B21/00 or F26B23/00
    • F26B25/06—Chambers, containers, or receptacles
    • F26B25/08—Parts thereof
    • F26B25/12—Walls or sides; Doors

Definitions

  • the invention relates to the field of cell preservation technology, and in particular to a cell freeze-drying system and method.
  • the commonly used cell dehydration method is a vacuum freeze-drying method, which relies on a vacuum freeze-drying device to realize the control of the gaseous water pressure in the confined space and the sublimation energy input, and complete the cell freeze-drying.
  • the lyophilized cells obtained by this method have obvious microstructural damage and the recovery rate is low.
  • the specific principle of freeze-dried damage is not clear, and the main influencing factor may be the internal stress of cells during lyophilization.
  • Working medium there is a huge temperature difference between the two, which may be an important cause of cell freeze-drying damage.
  • freeze-drying equipment commonly used in cell laboratories still has serious pollution problems. Since the device needs to continuously operate the vacuum pump for more than 24 hours to control the gaseous moisture pressure in the working space and maintain the negative pressure state, the vacuum pump continuously operates, the evaporation consumption of the oil cannot be avoided, and it is difficult to control severe vibration and noise. There is a problem of poor matching between the equipment and the cell laboratory, it is not possible to operate indoors, and it is difficult to connect with the cell freeze-thaw equipment.
  • Cells are the basic functional unit of complex living organisms, and the definition of human health and disease ultimately points to cells.
  • Human medicine began with the investigation of organs, systems, and tissues, and has now rapidly turned to cellular methods, including gene editing, high-throughput DNA sequencing, artificially modified immune cell methods, and so on. These methods can be referred to as clinical cell methods, and higher efficiency and lower cost clinical medical method systems are rapidly being established.
  • safe and low-cost preservation of cells has great social and economic significance.
  • Establishing safe, efficient, and low-cost cell lyophilization methods and equipment is one of the foundations of the cell medicine and cell industries.
  • the object of the embodiments of the present invention is to provide a cell freeze-drying system and method, which can effectively solve the problem of cell freeze-drying damage and pollution, and at the same time achieve safety, high efficiency and low cost.
  • an embodiment of the present invention provides a cell freeze-drying system, including:
  • Heating the nitrogen supply mechanism, the lyophilization work box and the venting mechanism wherein the warming nitrogen supply mechanism, the lyophilization work box and the venting mechanism are sequentially connected by a first connecting pipe; wherein the adding The warm nitrogen supply mechanism will heat the gas Nitrogen is supplied to the lyophilization work box; the lyophilization work box is heated in a preset mode after being placed in the cell sample while continuously feeding the heated gaseous nitrogen to freeze-dry the cell sample
  • the solid water of the cell sample is sublimated into gaseous water to the gaseous nitrogen, wherein the cell sample is a cell sample frozen by liquid nitrogen, and the freeze-drying work box is pre-adjusted to the appropriate The temperature at which the liquid nitrogen-frozen cell sample is placed; the venting mechanism is for discharging gaseous nitrogen containing gaseous water obtained by sublimation of solid water of the cell sample.
  • a cell lyophilization system disclosed by the present invention provides a heated nitrogen gas to a lyophilization work box by a heated nitrogen supply mechanism by providing a warm nitrogen supply mechanism, a lyophilization work box, and an exhaust mechanism. After the lyophilized work box has been pre-cooled, after the cell sample is placed, the temperature is raised according to a preset mode while continuing to pass the heated gaseous nitrogen, and the cell sample is freeze-dried to dry the clean gas state.
  • Nitrogen is used as a sublimation drying heat medium and an inert protective agent, and the liquid nitrogen temperature is used as a sublimation starting temperature, so that the solid water of the cell sample is sublimated into gaseous water, and then the solid water sublimation containing the cell sample is discharged through the exhaust mechanism.
  • the technical solution of the gaseous nitrogen of the gaseous water, the lyophilization work box has been pre-adjusted to a temperature suitable for the cell sample after the liquid nitrogen freezing, since the lyophilization work box has been pre-adjusted to the time when the cell sample is placed Suitable temperature, solves the problem of cell damage caused by excessive initial temperature difference.
  • Nitrogen is used as sublimation medium to provide sublimation energy through continuous heating. Can be done under normal pressure sublimation dehydration cell sample, and the price of industrial nitrogen, less energy consumption and environmental pollution, access to the safe, efficient and cost benefits.
  • freeze-drying work box comprises:
  • a sample access device for placing the cell sample in a lyophilization work box or taking the lyophilized cell sample from the lyophilization work box;
  • the tray is heated to warm the freeze-drying work box in a preset mode.
  • the warming nitrogen supply mechanism comprises:
  • liquid nitrogen storage tank for storing liquid nitrogen
  • a gas generating device for converting the liquid nitrogen into the gaseous nitrogen
  • a gas heating device for heating the gaseous nitrogen and supplying the heated gaseous nitrogen to the lyophilization work box
  • the liquid nitrogen tank, the gas generating device and the gas warming device are sequentially connected by a second connecting pipe, and the gas warming device is connected to the lyophilization working box through the first connecting pipe.
  • the lyophilization work box further includes a volume adjuster; the volume adjuster is for adjusting the capacity of the lyophilization work box.
  • the cell lyophilization system further includes a central controller and a flow sensor; the flow sensor is disposed at The lyophilization work box is passed to an inlet of the heated gaseous nitrogen, and the flow sensor is configured to obtain a flow rate of the heated gaseous nitrogen that is supplied, the central controller is configured to freeze according to the The capacity of the dry working tank and the flow rate of the gaseous nitrogen obtained by the flow sensor control the power of the gas generating device, the power of the gas warming device, and the thermal power of the heating plate to rise in a preset mode.
  • sample feeding device is further configured to acquire the weight of the sample to be placed
  • the central controller is further configured to control the volume adjuster to adjust a capacity of the lyophilization work box based on a weight of the cell sample obtained by the sample access device.
  • the exhaust mechanism further includes a heating device for controlling a rate at which the exhaust mechanism discharges the gaseous nitrogen by adjusting a temperature of the exhaust mechanism.
  • the embodiment of the invention further provides a cell lyophilization method, comprising:
  • the cell sample is placed in a lyophilization work box; wherein the cell sample is a cell sample that has been frozen by liquid nitrogen, and the lyophilization work box is pre-adjusted to a sample of the cell after freezing by liquid nitrogen. temperature;
  • Initiating a lyophilization process of the lyophilization work box comprising: heating the lyophilization work box according to a preset mode, and continuously passing the heated gaseous nitrogen into the lyophilization work box, so that the cell sample is included
  • the solid water is sublimated into gaseous water into the gaseous nitrogen while continuously discharging the gaseous nitrogen comprising gaseous water obtained by sublimation of solid water of the cell sample;
  • the lyophilization process of the lyophilization work box is ended and the cell sample is removed from the lyophilization work box.
  • a cell lyophilization method disclosed in the present invention comprises placing a cell sample frozen by liquid nitrogen into a lyophilization work box, and then starting a lyophilization process of the lyophilization work box, including lyophilization.
  • the working box is heated according to the preset mode, and the heated gaseous nitrogen is continuously introduced into the lyophilization working box, so that the solid water contained in the cell sample is sublimated into gaseous water, and the solid water containing the cell sample is continuously discharged.
  • Gaseous nitrogen in gaseous water when the preset freeze-drying stop condition is reached, the lyophilization process of the freeze-drying work box is terminated, and the cell sample is taken out from the lyophilization work box, and the lyophilized work box is placed as the cell sample is placed. It has been pre-adjusted to a suitable temperature to solve the problem of cell damage caused by excessive initial temperature difference. Nitrogen is used as a sublimation medium to provide sublimation energy by continuous heating, and sublimation dehydration of cell samples can be completed under normal pressure. Moreover, the price of nitrogen industry has less energy consumption for environmental pollution and has obtained the beneficial effects of safety, high efficiency and low cost.
  • the continuously feeding the heated gaseous nitrogen into the lyophilization work box comprises:
  • the heated gaseous nitrogen is passed to the lyophilization working chamber.
  • the placing the cell sample into the lyophilization work box specifically comprises:
  • the heating the lyophilization work box according to a preset mode specifically includes:
  • FIG. 1 is a schematic structural view of a cell lyophilization system according to Embodiment 1 of the present invention.
  • FIG. 2 is a schematic structural view of a cell lyophilization system according to Embodiment 2 of the present invention.
  • FIG. 3 is a schematic flow chart of a cell lyophilization method according to Embodiment 3 of the present invention.
  • FIG. 1 is a schematic structural diagram of a cell lyophilization system according to Embodiment 1 of the present invention, and an embodiment of the present invention includes:
  • the lyophilized working box 2 is heated in a preset mode after being placed in the cell sample, and continuously passes through the heated gaseous nitrogen to freeze-dry the cell sample, so that the solid water of the cell sample is sublimated into gaseous water to gaseous nitrogen.
  • the cell sample is a cell sample frozen by liquid nitrogen, and the lyophilization working box 2 has been previously adjusted to a temperature suitable for the cell sample to be frozen by liquid nitrogen.
  • the freeze-drying starting temperature of the existing equipment is generally higher than -90 ° C, and liquid nitrogen is commonly used as a working medium for cell freezing, and there is a huge temperature difference between the two, which easily causes cell freeze-drying damage; the suitable passing liquid referred to in the present invention Nitrogen-frozen cells
  • the temperature at which the sample is placed is a temperature which does not cause structural damage to the cell sample after freezing with liquid nitrogen, and an ideal temperature is preferably -186 °C.
  • the temperature of the actual freeze-drying work box 2 tends to be slightly higher than -186 ° C, but since the cell sample is under liquid nitrogen protection when entering the freeze-drying work box 2, the sample temperature is raised from the liquid nitrogen temperature to the temperature inside the box, and then frozen.
  • the dryness has been started, that is, the cell sample is still under the protection of liquid nitrogen at the beginning of lyophilization, and it can be considered that it is lyophilized from the liquid nitrogen temperature, and the liquid nitrogen temperature is the sublimation starting temperature.
  • the exhaust mechanism 3 is for discharging gaseous nitrogen containing gaseous water obtained by sublimation of solid water of a cell sample.
  • the lyophilization system of the embodiment of the invention is based on the use of dry clean gaseous nitrogen as a sublimation drying heat medium and an inert protective agent, and the liquid nitrogen temperature is used as the sublimation starting temperature, and the principle of sublimation dehydration of the cell sample is completed under normal pressure to realize the freezing of the cell sample. Dry processing. Specifically, nitrogen is introduced into the lyophilization work box. When the nitrogen gas is pure, it can be considered that the lyophilization work box does not contain water in the nitrogen environment, or the equilibrium of the sublimation water pressure is considered to be reached in the lyophilization work box at this time. After the temperature rises, the equilibrium state of nitrogen is broken, thereby starting the sublimation of the solid water in the cell sample, and the continuous heating can achieve continuous sublimation until the cells are completely dehydrated.
  • the first connecting pipes selected in the embodiment are all adiabatic connecting pipes, and the first connecting pipes are provided with one-way airtight valves to ensure the unidirectionality of gas circulation, thereby ensuring the normal operation of the freeze-drying treatment.
  • the freeze-drying work box 2 is started to start pre-cooling, and the warming nitrogen supply mechanism 1 and the exhaust mechanism 3 are started, so that the temperature of the freeze-drying work box 2 reaches a suitable cell sample after liquid nitrogen freezing.
  • the temperature is, for example, -186 ° C; then the cell sample is placed in the lyophilization work box 2; the lyophilization work box 2 is heated in a preset mode after being placed in the cell sample while continuing to pass the heated gaseous nitrogen to the cells
  • the sample is subjected to lyophilization treatment, and the solid water of the cell sample is sublimated into gaseous water to gaseous nitrogen.
  • the preset freeze-drying stop condition is reached, the cell sample is taken out from the lyophilization work box 2, and the lyophilization of the cell sample is completed. deal with.
  • the embodiment of the invention solves the problem of damage to the cell structure of the prior freeze-drying technology, and uses nitrogen as a medium for sublimation to consume less energy, and the industrial price of nitrogen is low, and the environmental pollution is small, and the safety is obtained. , efficient, low-cost benefits.
  • FIG. 2 is a schematic structural diagram of a cell lyophilization system according to Embodiment 2 of the present invention.
  • the embodiment of the present invention includes:
  • the lyophilized working box 2 is heated in a preset mode after being placed in the cell sample while continuously feeding the heated gaseous nitrogen to the pair.
  • the cell sample is subjected to lyophilization treatment, and the solid water of the cell sample is sublimated into gaseous water to gaseous nitrogen, wherein the cell sample is a cell sample frozen by liquid nitrogen, and the lyophilization work box 2 is pre-adjusted to a suitable liquid nitrogen freezing. After the cell sample is placed at the temperature.
  • the freeze-drying starting temperature of the existing equipment is generally higher than -90 ° C, and liquid nitrogen is commonly used as a working medium for cell freezing, and there is a huge temperature difference between the two, which easily causes cell freeze-drying damage; the suitable passing liquid referred to in the present invention
  • the nitrogen-frozen cell sample is placed at a temperature that does not cause structural damage to the cell sample after freezing with liquid nitrogen, and an ideal temperature is preferably -186 °C.
  • the temperature of the actual freeze-drying work box 2 tends to be slightly higher than -186 ° C, but since the cell sample is under liquid nitrogen protection when entering the freeze-drying work box 2, the sample temperature is raised from the liquid nitrogen temperature to the temperature inside the box, and then frozen.
  • the dryness has been started, that is, the cell sample is still under the protection of liquid nitrogen at the beginning of lyophilization, and it can be considered that it is lyophilized from the liquid nitrogen temperature, and the liquid nitrogen temperature is the sublimation starting temperature.
  • the exhaust mechanism 3 is for discharging gaseous nitrogen containing gaseous water obtained by sublimation of solid water of a cell sample.
  • the lyophilization system of the embodiment of the invention is based on the use of dry clean gaseous nitrogen as a sublimation drying heat medium and an inert protective agent, and the liquid nitrogen temperature is used as the sublimation starting temperature, and the principle of sublimation dehydration of the cell sample is completed under normal pressure to realize the freezing of the cell sample. Dry processing. Specifically, nitrogen is introduced into the lyophilization work box. When the nitrogen gas is pure, it can be considered that the lyophilization work box does not contain water in the nitrogen environment, or the equilibrium of the sublimation water pressure is considered to be reached in the lyophilization work box at this time. After the temperature rises, the equilibrium state of nitrogen is broken, thereby starting the sublimation of the solid water in the cell sample, and the continuous heating can achieve continuous sublimation until the cells are completely dehydrated.
  • the lyophilization work box 2 comprises:
  • the sample inlet and outlet device 21 is for placing a cell sample into the lyophilization work box 2 or taking out the lyophilized cell sample from the lyophilization work box 2.
  • the heating tray 22 is heated to warm the lyophilization working box 2 according to a preset mode; in addition, as shown in FIG. 1, the heating tray 22 can also serve as a carrier to place a cell sample placed in the lyophilization working box 2 through the sample inlet and outlet device 21. The cell sample is placed and the cell sample is transferred to the heating plate 22.
  • the heating plate 22 of the embodiment preferably uses a metal heating plate. Since the metal material has a larger heat capacity than nitrogen, the heating plate 22 made of a metal material can be used as the lyophilization work box 2.
  • the temperature-controlled buffer not only reduces the amount of nitrogen used, but also controls the temperature inside the freeze-drying work box 2.
  • the lyophilization work box 2 also preferably includes a volume adjuster 23 for adjusting the capacity of the lyophilization work box 2.
  • the warming nitrogen supply mechanism 1 comprises:
  • a liquid nitrogen storage tank 11 for storing liquid nitrogen for storing liquid nitrogen; preferably, a liquid nitrogen storage tank 11 equipped with a liquid nitrogen liquid level position sensor, It is used to know the liquid nitrogen storage or liquid nitrogen dosage of the liquid nitrogen storage tank 11.
  • a gas warming device 13 for heating gaseous nitrogen and supplying the heated gaseous nitrogen to the lyophilization work box;
  • the liquid nitrogen tank 11, the gas generating device 12, and the gas warming device 13 are sequentially connected by a second connecting pipe, and the gas warming device 13 is connected to the lyophilization working box 2 through a first connecting pipe.
  • the embodiment further includes a central controller 4 and a flow sensor 5; the flow sensor 5 is disposed at the inlet of the lyophilized working box 2 to the heated gaseous nitrogen, and the flow sensor 5 is used to obtain the access The flow rate of the heated gaseous nitrogen; the central controller is used to control the power of the gas generating device 12, the power of the gas warming device 13, and the heating plate according to the capacity of the lyophilization working box 2 and the flow rate of the gaseous nitrogen obtained by the flow sensor 5.
  • the thermal power of 22 rises according to the preset mode.
  • sample inlet and outlet device 21 is further configured to acquire the weight of the placed cell sample; the central controller 4 is further configured to adjust the volume adjuster 23 according to the weight of the cell sample obtained by the sample inlet and outlet device 21 to adjust the freeze-drying work box 2 capacity.
  • the exhaust mechanism 3 further includes a heating device 31 for controlling the rate at which the exhaust mechanism 31 discharges gaseous nitrogen by adjusting the temperature of the exhaust mechanism 3.
  • the heating device 31 can be controlled by the central controller 4.
  • the first connecting pipe and the second connecting pipe selected in this embodiment are all adiabatic connecting pipes, and a one-way airtight valve is arranged in the first connecting pipe and the second connecting pipe to ensure the unidirectionality of gas circulation. Thereby ensuring the normal operation of the freeze-drying treatment.
  • the first temperature sensor 61 in the gas heating device 13 the second temperature sensor 62 at the inlet of the lyophilization work box 2 into the gaseous nitrogen, the third temperature sensor 63 in the heating plate 22, and the heating device 31.
  • the temperature sensor 64 is so that the central controller 4 obtains the temperature values of the gas heating means 13, the inlet of the lyophilization working tank 2 into the gaseous nitrogen, the heating plate 22 and the heating means 31.
  • the central controller 4 is first activated, and the central controller 4 obtains the temperature values of the gas heating device 13, the inlet of the lyophilized working box 2 into the gaseous nitrogen, the heating plate 22 and the heating device 31;
  • the dry working box 2 starts pre-cooling, and starts the warming nitrogen supply mechanism 1 and the exhaust mechanism 3, where the gas generating device of the warming nitrogen supply mechanism 1 is activated, so that the temperature of the lyophilization working box 2 reaches a suitable liquid nitrogen.
  • the frozen cell sample is placed at a temperature, such as -186 ° C; then the cell sample is placed through the sample access device 21, the weight of the cell sample is obtained, and the cell sample is transferred to the heating plate 22; then, the central controller 4 is based on the cell
  • the weight of the sample is adjusted by the control volume adjuster 23 to adjust the capacity of the lyophilization work box 2, while the central controller 4 adjusts the power and gas of the gas generating device 12 according to the adjusted capacity of the freeze-drying work box 2 and the reading of the flow sensor 5.
  • the power of the warming device 13 and the thermal power of the heating plate 22 are in a preset mode.
  • the lyophilization work box is heated according to the preset mode while continuously feeding the heated gaseous nitrogen to initiate lyophilization of the cell sample; when the preset freeze-drying stop condition is reached, the cells are passed through the sample inlet and outlet device 21. The sample was taken out from the lyophilization work box 2, and the lyophilization treatment of the cell sample was completed.
  • the embodiment in which the air mechanism 3 implements the exhaust is a preferred embodiment, and it should be noted that those skilled in the art can implement the technical effect of the present invention by replacing the above embodiments without departing from the principle of the present invention. The improvement is also considered to be the scope of protection of the present invention.
  • the embodiment of the invention solves the problem of cell damage caused by excessive initial temperature difference, and uses nitrogen as a sublimation medium to provide sublimation energy by continuous heating, and can complete cell samples under normal pressure.
  • FIG. 3 is a schematic flowchart of a cell lyophilization method according to Embodiment 3 of the present invention, which specifically includes:
  • the cell sample is placed in a lyophilization working box; wherein, the cell sample is a cell sample frozen by liquid nitrogen, and the lyophilization working box is pre-adjusted to a temperature suitable for the cell sample after freezing by liquid nitrogen;
  • starting the freeze-drying process of the freeze-drying work box comprising: heating the freeze-drying work box according to a preset mode, and continuously passing the heated gaseous nitrogen into the freeze-drying work box, so that the solid water contained in the cell sample is sublimated into a gaseous state. Water, while continuously discharging gaseous nitrogen containing gaseous water sublimated from solid water of the cell sample;
  • the lyophilization system of the embodiment of the invention is based on the use of dry clean gaseous nitrogen as a sublimation drying heat medium and an inert protective agent, and the liquid nitrogen temperature is used as the sublimation starting temperature, and the principle of sublimation dehydration of the cell sample is completed under normal pressure to realize the freezing of the cell sample. Dry processing. Specifically, nitrogen is introduced into the lyophilization work box. When the nitrogen is pure, it can be considered that the nitrogen atmosphere of the freeze-drying work box does not contain water at this time, or that the sublimation water pressure is reached in the freeze-drying work box at this time. Equilibrium; after the temperature rises, the equilibrium state of nitrogen is broken, thereby initiating sublimation of solid water in the cell sample, and continuous heating can achieve continuous sublimation to complete dehydration of the cells.
  • the existing freeze-drying starting point temperature is generally higher than -90 ° C, while the cell freezing commonly uses liquid nitrogen as a working medium, both of which are stored.
  • the temperature of the cell sample which is suitable for freezing by liquid nitrogen in the present invention is a temperature which does not cause structural damage to the cell sample which has been frozen by liquid nitrogen, and is ideal.
  • a suitable temperature is -186 °C.
  • the pre-cooling temperature of the actual lyophilization work box tends to be slightly higher than -186 ° C, but since the cell sample is under liquid nitrogen protection when entering the lyophilization work box, the sample temperature rises from the liquid nitrogen temperature to the temperature inside the chamber. Freeze-drying has been initiated, that is, the cell sample is still under the protection of liquid nitrogen at the beginning of lyophilization, which can be considered as lyophilization from liquid nitrogen temperature, and the liquid nitrogen temperature is the sublimation starting temperature.
  • the purity of the nitrogen obtained by direct gasification of liquid nitrogen is high, and the continuous passage of the heated gaseous nitrogen into the lyophilization working chamber in the lyophilization step in step S2 is preferably: first converting liquid nitrogen into gaseous nitrogen to obtain gaseous nitrogen. The gaseous nitrogen is then heated; finally, the heated gaseous nitrogen is passed to the lyophilization working chamber.
  • first converting liquid nitrogen into gaseous nitrogen to obtain gaseous nitrogen.
  • the gaseous nitrogen is then heated; finally, the heated gaseous nitrogen is passed to the lyophilization working chamber.
  • an embodiment in which other embodiments are converted based on the principle of the present invention to obtain heated nitrogen gas is also considered as the protection scope of the present invention.
  • the temperature is raised according to the preset mode, and it is preferred that the gaseous nitrogen is also heated according to the preset mode, so that the temperature of the gaseous nitrogen flowing into the lyophilization work box can be substantially frozen.
  • the dry working chamber temperature is consistent, providing continuous energy for the sublimation of solid water in the cell sample.
  • the gaseous nitrogen containing the gaseous water obtained by sublimating the solid water of the cell sample in the step S2 can be preferably heated to force the gaseous nitrogen containing the gaseous water obtained by sublimating the solid water of the cell sample. Discharge, improve exhaust efficiency.
  • step S3 specifically includes:
  • the cell sample is placed in a pre-cooled lyophilization work box, and the weight of the cell sample is obtained, and the capacity of the lyophilization work box is adjusted according to the weight of the cell sample;
  • step S2 the lyophilization work box is heated according to the preset mode, and specifically includes:
  • the cell sample frozen by liquid nitrogen is first placed in a lyophilization work box, and the lyophilization work box is pre-adjusted to a temperature suitable for the cell sample after freezing by liquid nitrogen; then the freeze-drying work box is started.
  • the lyophilization process comprises: heating the lyophilized work box according to a preset mode, and continuously passing the heated gaseous nitrogen into the lyophilization work box, so that the solid water contained in the cell sample is sublimated into gaseous water, and continuously discharged The gaseous nitrogen of the gaseous water obtained by sublimating the solid water of the cell sample; when the preset freeze-drying stop condition is reached, the lyophilization process of the freeze-drying work box is ended, and the cell sample is taken out from the lyophilization work box.
  • the embodiment of the invention solves the problem of damage to the cell structure by the prior freeze-drying technology, adopts nitrogen as a medium for sublimation, consumes less energy, and has low industrial price of nitrogen and little environmental pollution, and obtains Safe, efficient and low cost benefits.

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Abstract

一种细胞冻干系统，包括：加温氮气供给机构、冻干工作箱和排气机构，所述加温氮气供给机构、所述冻干工作箱和所述排气机构之间依次通过第一连接管连接；所述加温氮气供给机构将加热后的气态氮供给至所述冻干工作箱；所述冻干工作箱在放入经过液氮冷冻后的细胞样品后按照预设模式升温同时持续通入加热后的气态氮，使细胞样品的固态水升华为气态水；所述冻干工作箱已预先调整至适宜经过液氮冷冻后的细胞样品放入的温度；所述排气机构用于排出包含由细胞样品的固态水升华而得的气态水的气态氮。还涉及一种细胞冻干方法。能有效解决细胞冻干损伤和污染的问题，实现细胞冻干的安全、高效和低成本。

Description

一种细胞冻干系统和方法 技术领域
本发明涉及细胞保存技术领域，尤其涉及一种细胞冻干系统和方法。
背景技术
常用的细胞脱水方法为真空冻干法，该方法依赖真空冻干装置实现局限空间内气态水分压的控制和升华能源输入，完成细胞冻干。这一方法获得的冻干细胞存在明显的微结构破坏，复苏率偏低。冻干损伤的具体原理不明确，主要的影响因素可能是冻干过程中细胞内部应力作用。目前常用的工业设备条件下，难以低成本获得低于-90℃的可控温度环境，用于细胞冻干的人工环境起始工作温度一般不低于-90℃，而细胞冷冻常用液氮作为工作介质，二者存在巨大的温度差异，这可能是导致细胞冻干损伤的重要原因。
目前，细胞实验室常用的冻干设备还存在严重的污染问题。由于该设备需要真空泵连续运转超过24小时，以控制工作空间的气态水分压、维持负压状态，真空泵连续运转，不能避免滑油的蒸发消耗，也很难控制严重的振动与噪音，因此，该设备与细胞实验室存在匹配不良的问题，不能室内运行，也难以与细胞冻融设备进行工序连接。
细胞是复杂生命体基本的功能单元，人类健康与疾病的定义对象最终都指向细胞。人类医学从以器官、系统、组织为考察和干预对象开始，现在已经快速转向到细胞方法，包括基因编辑、高通量DNA测序、人工修饰的免疫细胞方法，等等。这些方法可以称为临床细胞方法，更高效率更低成本的临床医学方法体系正在快速建立。细胞医学时代，安全、低成本保存细胞，具有重大的社会意义和经济意义。建立安全、高效、低成本的细胞冻干方法和设备，是细胞医学和细胞工业的基础之一。
发明内容
本发明实施例的目的是提供一种细胞冻干系统和方法，能有效解决细胞冻干损伤和污染的问题，同时实现安全、高效、低成本。
为实现上述目的，本发明实施例提供了一种细胞冻干系统，包括：
加温氮气供给机构、冻干工作箱和排气机构，所述加温氮气供给机构、所述冻干工作箱和所述排气机构之间依次通过第一连接管连接；其中，所述加温氮气供给机构将加热后的气 态氮供给至所述冻干工作箱；所述冻干工作箱在放入细胞样品后按照预设模式升温同时持续通入加热后的所述气态氮，以对所述细胞样品进行冻干处理，使所述细胞样品的固态水升华为气态水至所述气态氮中，其中，所述细胞样品为经过液氮冷冻后的细胞样品，所述冻干工作箱已预先调整至适宜所述经过液氮冷冻后的细胞样品放入的温度；所述排气机构用于排出包含由所述细胞样品的固态水升华而得的气态水的气态氮。
与现有技术相比，本发明公开的一种细胞冻干系统通过设置加温氮气供给机构、冻干工作箱和排气机构，由加热氮气供给机构提供加热后的氮气至冻干工作箱，由冻干工作箱已预先预冷的情况下在放入细胞样品后，按照预设模式升温同时持续通入加热后的所述气态氮，对所述细胞样品进行冻干处理，以干燥洁净气态氮作为升华干燥的热介质与惰性保护剂，以液氮温度为升华起点温度，使所述细胞样品的固态水升华为气态水，然后通过排气机构排出包含由所述细胞样品的固态水升华而得的气态水的气态氮的技术方案，冻干工作箱已预先调整至适宜所述经过液氮冷冻后的细胞样品放入的温度，由于细胞样品放入时冻干工作箱已预先调整至适宜温度，解决了由于放入的初始温差过大而导致的细胞损伤问题，采用氮作为升华介质，通过持续升温提供升华能量，能够在常压状态下完成细胞样品的升华脱水，而且氮工业价格，对环境污染小耗能少，获得了安全、高效、低成本的有益效果。
进一步的，所述冻干工作箱包括：
样品进出装置，用于将所述细胞样品放入冻干工作箱内或将所述冻干处理完成的细胞样品从所述冻干工作箱取出；
加热盘，使所述冻干工作箱按照预设模式升温。
进一步的，所述加温氮气供给机构包括：
液氮储箱，用于储存液态氮；
气体发生装置，用于将所述液态氮转化为所述气态氮；
气体加温装置，用于加热所述气态氮，并将加热后的所述气态氮供给至所述冻干工作箱；
所述液氮储箱、所述气体发生装置和所述气体加温装置依次通过第二连接管连接，所述气体加温装置通过所述第一连接管与所述冻干工作箱连接。
进一步的，所述冻干工作箱还包括容积调节器；所述容积调节器用于调节所述冻干工作箱的容量。
进一步的，所述细胞冻干系统还包括中央控制器和流量传感器；所述流量传感器设置于 所述冻干工作箱通入加热后的所述气态氮的入口处，所述流量传感器用于获取所通入的加热后的所述气态氮的流量，所述中央控制器用于根据所述冻干工作箱的容量和所述流量传感器获取的所述气态氮的流量控制所述气体发生装置的功率、所述气体加温装置的功率和所述加热盘的热功率按照预设模式上升。
进一步的，所述样品进出装置还用于获取放入的将所述细胞样品的重量；
所述中央控制器还用于根据所述样品进出装置获取的所述细胞样品的重量控制所述容积调节器调整所述冻干工作箱的容量。
进一步的，所述排气机构还包括加热装置；所述加热装置用于通过调整所述排气机构的温度来控制所述排气机构排出所述气态氮的速率。
相应的，本发明实施例还提供一种细胞冻干方法，包括：
将细胞样品放入冻干工作箱中；其中，所述细胞样品为经过液氮冷冻后的细胞样品，所述冻干工作箱已预先调整至适宜所述经过液氮冷冻后的细胞样品放入的温度；
启动所述冻干工作箱的冻干工序，包括使所述冻干工作箱按照预设模式进行升温，并持续将加热后的气态氮通入所述冻干工作箱，使所述细胞样品包含的固态水升华为气态水至所述气态氮中，同时不断排出包含由所述细胞样品的固态水升华而得的气态水的所述气态氮；
当达到预设冻干停止条件时，结束所述冻干工作箱的冻干工序，并将所述细胞样品从所述冻干工作箱取出。
与现有技术相比，本发明公开的一种细胞冻干方法通过将经过液氮冷冻后的细胞样品放入冻干工作箱中，然后启动冻干工作箱的冻干工序，包括使冻干工作箱按照预设模式进行升温，并持续将加热后的气态氮通入冻干工作箱，使细胞样品包含的固态水升华为气态水，同时不断排出包含由细胞样品的固态水升华而得的气态水的气态氮；当达到预设冻干停止条件时，结束冻干工作箱的冻干工序，并将细胞样品从冻干工作箱取出的技术方案，由于细胞样品放入时冻干工作箱已预先调整至适宜温度，解决了由于放入的初始温差过大而导致的细胞损伤问题，采用氮作为升华介质，通过持续升温提供升华能量，能够在常压状态下完成细胞样品的升华脱水，而且氮工业价格，对环境污染小耗能少，获得了安全、高效、低成本的有益效果。
进一步的，所述持续将加热后的气态氮通入所述冻干工作箱具体包括：
将液态氮转化为所述气态氮，从而获取所述气态氮；
对所述气态氮进行加热；
将经过加热的所述气态氮通入所述冻干工作箱。
进一步的，所述将细胞样品放入冻干工作箱中具体包括：
将细胞样品放入预冷后的所述冻干工作箱，并获取所述细胞样品的重量，根据所述细胞样品的重量调整所述冻干工作箱的容量；
所述使所述冻干工作箱按照预设模式进行升温具体包括：
获取通入所述冻干工作箱的所述气态氮的流量，并根据所述冻干工作箱的容量和通入所述冻干工作箱的所述气态氮的流量，控制所述冻干工作箱按照预设模式进行升温。
附图说明
图1是本发明实施例1提供的一种细胞冻干系统的结构示意图；
图2是本发明实施例2提供的一种细胞冻干系统的结构示意图；
图3是本发明实施例3提供的一种细胞冻干方法的流程示意图。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
参见图1，是本发明实施1提供的一种细胞冻干系统的结构示意图，本发明实施例包括：
加温氮气供给机构1、冻干工作箱2和排气机构3，其中，加温氮气供给机构1、冻干工作箱2和排气机构3之间依次通过第一连接管连接，具体的：
加温氮气供给机构1将加热后的气态氮供给至冻干工作箱2；
冻干工作箱2在放入细胞样品后按照预设模式升温同时持续通入加热后的气态氮，以对细胞样品进行冻干处理，使细胞样品的固态水升华为气态水至气态氮中，其中，细胞样品为经过液氮冷冻后的细胞样品，冻干工作箱2已预先调整至适宜经过液氮冷冻后的细胞样品放入的温度。
通常，现有设备的冻干起点温度一般高于-90℃，而细胞冷冻常用液氮作为工作介质，二者存在巨大的温度差异，容易导致细胞冻干损伤；本发明所指的适宜经过液氮冷冻后的细胞 样品放入的温度为不会对经过液氮冷冻后的细胞样品造成结构损伤的温度，理想的适宜温度为-186℃。实际冻干工作箱2的温度往往稍高于-186℃，但是由于细胞样品在进入冻干工作箱2时是在液氮保护下，样品温度从液氮温度升高到箱内温度前，冻干已经启动了，即冻干开始时细胞样品还是在液氮保护下，可以认为就是从液氮温度冻干，以液氮温度为升华起点温度。
排气机构3用于排出包含由细胞样品的固态水升华而得的气态水的气态氮。
本发明实施例的冻干系统基于使用干燥洁净气态氮作为升华干燥的热介质与惰性保护剂，以液氮温度为升华起点温度，常压状态下完成细胞样品升华脱水的原理实现细胞样品的冻干处理。具体的，在冻干工作箱中通入氮气，在氮气为纯净的情况下，可以认为此时冻干工作箱氮气环境中不含水，或者认为此时冻干工作箱内达到升华水分压的平衡态；在温度升高后，打破了氮气的平衡态，从而启动细胞样品中的固态水的升华，持续的加温可实现持续的升华至细胞完全脱水。
本实施例所选用的第一连接管均为绝热连接管，在第一连接管中均设有单向气密阀，以保证气体流通的单向性，从而保证冻干处理的正常进行。
具体实施时，首先启动冻干工作箱2开始进行预冷，以及启动加温氮气供给机构1和排气机构3，使冻干工作箱2的温度达到适宜经过液氮冷冻后的细胞样品放入的温度，如-186℃；然后在冻干工作箱2中放入细胞样品；冻干工作箱2在放入细胞样品后按照预设模式升温同时持续通入加热后的气态氮，以对细胞样品进行冻干处理，使细胞样品的固态水升华为气态水至气态氮中，在达到预设冻干停止条件时，将细胞样品从冻干工作箱2取出，完成了对细胞样品的冻干处理。
与现有技术相比，本发明实施例解决了现有冻干技术对细胞结构的破坏问题，采用氮作为介质进行升华耗能少，而且氮的工业价格低，对环境污染小，获得了安全、高效、低成本的有益效果。
参见图2，是本发明实施2提供的一种细胞冻干系统的结构示意图，本发明实施例包括：
加温氮气供给机构1、冻干工作箱2和排气机构3，其中，加温氮气供给机构1、冻干工作箱2和排气机构3之间依次通过第一连接管连接，具体的：
加温氮气供给机构1将加热后的气态氮供给至冻干工作箱2；
冻干工作箱2在放入细胞样品后按照预设模式升温同时持续通入加热后的气态氮，以对 细胞样品进行冻干处理，使细胞样品的固态水升华为气态水至气态氮中，其中，细胞样品为经过液氮冷冻后的细胞样品，冻干工作箱2已预先调整至适宜经过液氮冷冻后的细胞样品放入的温度。
通常，现有设备的冻干起点温度一般高于-90℃，而细胞冷冻常用液氮作为工作介质，二者存在巨大的温度差异，容易导致细胞冻干损伤；本发明所指的适宜经过液氮冷冻后的细胞样品放入的温度为不会对经过液氮冷冻后的细胞样品造成结构损伤的温度，理想的适宜温度为-186℃。实际冻干工作箱2的温度往往稍高于-186℃，但是由于细胞样品在进入冻干工作箱2时是在液氮保护下，样品温度从液氮温度升高到箱内温度前，冻干已经启动了，即冻干开始时细胞样品还是在液氮保护下，可以认为就是从液氮温度冻干，以液氮温度为升华起点温度。
排气机构3用于排出包含由细胞样品的固态水升华而得的气态水的气态氮。
本发明实施例的冻干系统基于使用干燥洁净气态氮作为升华干燥的热介质与惰性保护剂，以液氮温度为升华起点温度，常压状态下完成细胞样品升华脱水的原理实现细胞样品的冻干处理。具体的，在冻干工作箱中通入氮气，在氮气为纯净的情况下，可以认为此时冻干工作箱氮气环境中不含水，或者认为此时冻干工作箱内达到升华水分压的平衡态；在温度升高后，打破了氮气的平衡态，从而启动细胞样品中的固态水的升华，持续的加温可实现持续的升华至细胞完全脱水。
优选的，冻干工作箱2包括：
样品进出装置21，用于将细胞样品放入冻干工作箱2内或将冻干处理完成的细胞样品从冻干工作箱2取出。
加热盘22，使冻干工作箱2按照预设模式升温；另外，如图1所示，加热盘22还可以作为承载器来放置放入冻干工作箱2的细胞样品，通过样品进出装置21放入细胞样品并将细胞样品传送至加热盘22，本实施例的加热盘22优选采用金属加热盘，由于金属材料热容量较氮气大，采用金属材料的加热盘22可以作为冻干工作箱2内的温度控制的缓冲器，不仅能减少氮气用量，且易于控制冻干工作箱2内温度。
另外，冻干工作箱2还优选包括容积调节器23；容积调节器23用于调节冻干工作箱2的容量。
优选的，加温氮气供给机构1包括：
液氮储箱11，用于储存液态氮；优选，采用安装有液氮液面位置传感器的液氮储箱11， 用于获知液氮储箱11的液氮储量或液氮用量。
气体发生装置12，用于将液态氮转化为气态氮；
气体加温装置13，用于加热气态氮，并将加热后的气态氮供给至冻干工作箱；
液氮储箱11、气体发生装置12和气体加温装置13依次通过第二连接管连接，气体加温装置13通过第一连接管与冻干工作箱2连接。
为实现自动化和智能化，本实施例还包括中央控制器4和流量传感器5；流量传感器5设置于冻干工作箱2通入加热后的气态氮的入口处，流量传感器5用于获取所通入的加热后的气态氮的流量；中央控制器用于根据冻干工作箱2的容量和流量传感器5获取的气态氮的流量控制气体发生装置12的功率、气体加温装置13的功率和加热盘22的热功率按照预设模式上升。
进一步的，样品进出装置21还用于获取放入的将细胞样品的重量；中央控制器4还用于根据样品进出装置21获取的细胞样品的重量控制容积调节器23调整冻干工作箱2的容量。
进一步的，排气机构3还包括加热装置31；加热装置31用于通过调整排气机构3的温度来控制排气机构31排出气态氮的速率。优选的，加热装置31可以通过中央控制器4进行控制。
本实施例所选用的第一连接管和第二连接管均为绝热连接管，在第一连接管和第二连接管中均设有单向气密阀，以保证气体流通的单向性，从而保证冻干处理的正常进行。
另外，优选在气体加热装置13设置第一温度传感器61，冻干工作箱2通入气态氮的入口处设置第二温度传感器62，加热盘22设置第三温度传感器63，以及加热装置31设置第四温度传感器64，以便中央控制器4获取气体加热装置13、冻干工作箱2通入气态氮的入口处、加热盘22和加热装置31各处的温度值。
具体实施时，首先启动中央控制器4，中央控制器4获取气体加热装置13、冻干工作箱2通入气态氮的入口处、加热盘22和加热装置31各处的温度值；然后启动冻干工作箱2开始进行预冷，以及启动加温氮气供给机构1和排气机构3，此处启动加温氮气供给机构1的气体发生装置，使冻干工作箱2的温度达到适宜经过液氮冷冻后的细胞样品放入的温度，如-186℃；然后通过样品进出装置21放入细胞样品，获取细胞样品的重量，并将细胞样品传送至加热盘22；接着，中央控制器4依据细胞样品的重量通过控制容积调节器23调整冻干工作箱2的容量，同时，中央控制器4依据调整后的冻干工作箱2的容量与流量传感器5的读数调整气体发生装置12的功率、气体加温装置13的功率和加热盘22的热功率按照预设模式 上升，以使冻干工作箱按照预设模式升温同时持续通入加热后的气态氮，以启动对细胞样品进行冻干处理；在达到预设冻干停止条件时，通过样品进出装置21将细胞样品从冻干工作箱2取出，完成了对细胞样品的冻干处理。
本实施例中所提及的中央控制器4对冻干工作箱2的温度模式的调节和对容量的调节的实施方式、加温氮气供给机构1如何获取加热后的气态氮的实施方式，排气机构3如何实现排气的实施方式为优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，替换上述实施方式而实现本发明的技术效果的改进方案也视为本发明的保护范围。
与现有技术相比，本发明实施例解决了由于放入的初始温差过大而导致的细胞损伤问题，采用氮作为升华介质，通过持续升温提供升华能量，能够在常压状态下完成细胞样品的升华脱水，而且氮工业价格，对环境污染小耗能少，获得了安全、高效、低成本的有益效果；同时本实施例的冻干系统为完全可控的冻干环境，只需通过合理设置程序即可保证工作效率。
相应的，参见图3，图3是本发明实施例3提供的一种细胞冻干方法的流程示意图，具体包括：
S1、将细胞样品放入冻干工作箱中；其中，细胞样品为经过液氮冷冻后的细胞样品，冻干工作箱已预先调整至适宜经过液氮冷冻后的细胞样品放入的温度；
S2、启动冻干工作箱的冻干工序，包括使冻干工作箱按照预设模式进行升温，并持续将加热后的气态氮通入冻干工作箱，使细胞样品包含的固态水升华为气态水，同时不断排出包含由细胞样品的固态水升华而得的气态水的气态氮；
S3、当达到预设冻干停止条件时，结束冻干工作箱的冻干工序，并将细胞样品从冻干工作箱取出。
本发明实施例的冻干系统基于使用干燥洁净气态氮作为升华干燥的热介质与惰性保护剂，以液氮温度为升华起点温度，常压状态下完成细胞样品升华脱水的原理实现细胞样品的冻干处理。具体的，在冻干工作箱中通入氮气，在氮气为纯净的情况下，可以认为此时冻干工作箱的氮气环境中不含水，或者认为此时冻干工作箱内达到升华水分压的平衡态；在温度升高后，打破了氮气的平衡态，从而启动细胞样品中的固态水的升华，持续的加温可实现持续的升华至细胞完全脱水。
通常，现有的冻干起点温度一般高于-90℃，而细胞冷冻常用液氮作为工作介质，二者存 在巨大的温度差异，容易导致细胞冻干损伤；本发明所指的适宜经过液氮冷冻后的细胞样品放入的温度为不会对经过液氮冷冻后的细胞样品造成结构损伤的温度，理想的适宜温度为-186℃。实际冻干工作箱的预冷的温度往往稍高于-186℃，但是由于细胞样品在进入冻干工作箱时是在液氮保护下，样品温度从液氮温度升高到箱内温度前，冻干已经启动了，即冻干开始时细胞样品还是在液氮保护下，可以认为就是从液氮温度冻干，以液氮温度为升华起点温度。
由液氮直接气化所获得氮气的纯度较高，步骤S2冻干工序中的持续将加热后的气态氮通入冻干工作箱优选为：首先将液态氮转化为气态氮，从而获取气态氮；然后对气态氮进行加热；最后将经过加热的气态氮通入冻干工作箱。此处为优选实施方式，由于如何获取氮气以及对氮气加热均可通过现有技术获得，基于本发明原理转换其他实施方式来获取加热后的氮气的实施方式也视为本发明的保护范围。
另外，为保证冻干工作箱启动冻干工序时按照预设模式进行升温，优选对通入气态氮也按照预设模式进行升温，使通入冻干工作箱的气态氮的温度能够基本与冻干工作箱温度保持一致，为细胞样品中的固态水的升华持续提供能量。
步骤S2冻干工序中的排出包含由细胞样品的固态水升华而得的气态水的气态氮可以优选采用加热的方式，强制使包含由细胞样品的固态水升华而得的气态水的气态氮能够排出，提高排气效率。
步骤S3中将细胞样品放入冻干工作箱中具体包括：
将细胞样品放入预冷后的冻干工作箱，并获取细胞样品的重量，根据细胞样品的重量调整冻干工作箱的容量；
对应的，步骤S2中使冻干工作箱按照预设模式进行升温具体包括：
获取通入冻干工作箱的气态氮的流量，并根据冻干工作箱的容量和通入冻干工作箱的气态氮的流量，控制冻干工作箱按照预设模式进行升温。
具体实施时，首先将经过液氮冷冻后的细胞样品放入冻干工作箱中，冻干工作箱已预先调整至适宜经过液氮冷冻后的细胞样品放入的温度；然后启动冻干工作箱的冻干工序，包括使冻干工作箱按照预设模式进行升温，并持续将加热后的气态氮通入冻干工作箱，使细胞样品包含的固态水升华为气态水，同时不断排出包含由细胞样品的固态水升华而得的气态水的气态氮；当达到预设冻干停止条件时，结束冻干工作箱的冻干工序，并将细胞样品从冻干工作箱取出。
与现有技术相比，本发明实施例解决了现有冻干技术对细胞结构的破坏问题，采用氮作为介质进行升华，耗能少，而且氮的工业价格低，对环境污染小，获得了安全、高效、低成本的有益效果。
以上所述是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也视为本发明的保护范围。

Claims (10)

  1. 一种细胞冻干系统，其特征在于，包括：加温氮气供给机构、冻干工作箱和排气机构，所述加温氮气供给机构、所述冻干工作箱和所述排气机构之间依次通过第一连接管连接；其中，所述加温氮气供给机构将加热后的气态氮供给至所述冻干工作箱；所述冻干工作箱在放入细胞样品后按照预设模式升温同时持续通入加热后的所述气态氮，以对所述细胞样品进行冻干处理，使所述细胞样品的固态水升华为气态水，其中，所述细胞样品为经过液氮冷冻后的细胞样品，所述冻干工作箱已预先调整至适宜所述经过液氮冷冻后的细胞样品放入的温度；所述排气机构用于排出包含由所述细胞样品的固态水升华而得的气态水的气态氮。
  2. 如权利要求1所述的一种细胞冻干系统，其特征在于，所述冻干工作箱包括：
    样品进出装置，用于将所述细胞样品放入冻干工作箱内或将所述冻干处理完成的细胞样品从所述冻干工作箱取出；
    加热盘，使所述冻干工作箱按照预设模式升温。
  3. 如权利要求1所述的一种细胞冻干系统，其特征在于，所述加温氮气供给机构包括：
    液氮储箱，用于储存液态氮；
    气体发生装置，用于将所述液态氮转化为所述气态氮；
    气体加温装置，用于加热所述气态氮，并将加热后的所述气态氮供给至所述冻干工作箱；
    所述液氮储箱、所述气体发生装置和所述气体加温装置依次通过第二连接管连接，所述气体加温装置通过所述第一连接管与所述冻干工作箱连接。
  4. 如权利要求2所述的一种细胞冻干系统，其特征在于，所述冻干工作箱还包括容积调节器；所述容积调节器用于调节所述冻干工作箱的容量。
  5. 如权利要求4所述的一种细胞冻干系统，其特征在于，所述细胞冻干系统还包括中央控制器、流量传感器；所述流量传感器设置于所述冻干工作箱通入加热后的所述气态氮的入 口处；所述流量传感器用于获取所通入的加热后的所述气态氮的流量；所述中央控制器用于根据所述冻干工作箱的容量和所述流量传感器获取的所述气态氮的流量控制所述气体发生装置的功率、所述气体加温装置的功率和所述加热盘的热功率按照预设模式上升。
  6. 如权利要求5所述的一种细胞冻干系统，其特征在于，所述样品进出装置还用于获取放入的将所述细胞样品的重量；
    所述中央控制器还用于根据所述样品进出装置获取的所述细胞样品的重量控制所述容积调节器调整所述冻干工作箱的容量。
  7. 如权利要求5所述的一种细胞冻干系统，其特征在于，所述排气机构还包括加热装置；所述加热装置用于通过调整所述排气机构的温度来控制所述排气机构排出所述气态氮的速率。
  8. 一种细胞冻干方法，其特征在于，包括：
    将细胞样品放入冻干工作箱中；其中，所述细胞样品为经过液氮冷冻后的细胞样品，所述冻干工作箱已预先调整至适宜所述经过液氮冷冻后的细胞样品放入的温度；
    启动所述冻干工作箱的冻干工序，包括使所述冻干工作箱按照预设模式进行升温，并持续将加热后的气态氮通入所述冻干工作箱，使所述细胞样品包含的固态水升华为气态水至所述气态氮中，同时不断排出包含由所述细胞样品的固态水升华而得的气态水的所述气态氮；
    当达到预设冻干停止条件时，结束所述冻干工作箱的冻干工序，并将所述细胞样品从所述冻干工作箱取出。
  9. 如权利要求8所述的一种细胞冻干方法，其特征在于，所述持续将加热后的气态氮通入所述冻干工作箱具体包括：
    将液态氮转化为所述气态氮，从而获取所述气态氮；
    对所述气态氮进行加热；
    将经过加热的所述气态氮通入所述冻干工作箱。
  10. 如权利要求9所述的一种细胞冻干方法，其特征在于，所述将细胞样品放入冻干工作箱中具体包括：
    将细胞样品放入预冷后的所述冻干工作箱，并获取所述细胞样品的重量，根据所述细胞样品的重量调整所述冻干工作箱的容量；
    所述使所述冻干工作箱按照预设模式进行升温具体包括：
    获取通入所述冻干工作箱的所述气态氮的流量，并根据所述冻干工作箱的容量和通入所述冻干工作箱的所述气态氮的流量，控制所述冻干工作箱按照预设模式进行升温。
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