CN110742057B

CN110742057B - Low-temperature preservation method for autologous fat particles

Info

Publication number: CN110742057B
Application number: CN201910955292.7A
Authority: CN
Inventors: 张红芳; 刘中国; 张丽
Original assignee: Beijing Runmei Yuzhiguang Medical Beauty Clinic
Current assignee: Beijing Runmei Yuzhiguang Medical Beauty Clinic
Priority date: 2019-10-09
Filing date: 2019-10-09
Publication date: 2021-10-29
Anticipated expiration: 2039-10-09
Also published as: CN110742057A

Abstract

The invention relates to a low-temperature preservation method of autologous fat particles, which comprises the following steps: firstly, loading the fat particles after purification, cleaning and filtration into a sealed container (2); thirdly, the sealed container (2) filled with the fat particles is placed into the low-temperature preservation equipment provided by the application, and the freezing and cooling are started; fourthly, in the initial stage of freezing and cooling, the internal space of the sealed container (2) is cleaned by gas; fifthly, filling the gas into the sealed container (2) after the gas cleaning step and before the fat completely enters the frozen state; and sixthly, after the fat is completely in the frozen state, closing the gas pump for inputting gas and the gas pump for extracting gas by the controller (13), and closing the electromagnetic valves positioned on the gas conveying branch pipe (33) and the gas discharge branch pipe (34) to stabilize the atmosphere inside the sealed container (2) and enter a stable atmosphere frozen state.

Description

Low-temperature preservation method for autologous fat particles

Technical Field

The invention relates to the technical field of biology, in particular to a low-temperature preservation method for autologous fat particles.

Background

Autologous fat transplantation techniques have been widely used, for example: enlarging breast, enlarging nose or repairing scar. Compared with the traditional artificial tissue substitute, the autologous fat has the advantages of good compatibility, simple and convenient operation, good filling appearance, small surgical wound, rich source and the like. However, the prior art still has the problems of low survival rate of transplanted fat cells, necrosis, absorption, infection and the like, and the transplantation often needs a small amount of repeated injections. Therefore, how to improve the survival rate of the low-temperature preservation of the autologous fat particles is an urgent problem to be solved.

Currently, cryopreservation of fat particles tends to store the fat particles in a cryoprotectant and provide lower temperatures, such as: -80 degrees celsius for storage. However, the atmosphere environment of the storage space is often neglected in the existing storage method, and the performance of fat storage is cracked due to poor atmosphere environment of the storage space.

Therefore, it is desirable to provide a method for cryopreserving autologous fat particles, which can prevent fat contamination and improve the survival rate of later fat transplantation.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: provides a method for preserving autologous fat particles at a low temperature, which can avoid fat pollution and improve the survival rate of later fat transplantation.

The technical scheme adopted by the invention for solving the technical problems is as follows:

a low-temperature preservation device for autologous fat particles comprises a refrigerated cabinet 1, a bearing plate 21 positioned in the refrigerated cabinet 1, an air path system, a sealed container 2 and a controller 13; the refrigerated cabinet 1 comprises an accommodating space and a gas supply cavity, the accommodating space forms a plurality of independent spaces for accommodating the sealed containers 2 through a plurality of clapboards 11, and the gas path system comprises a gas conveying main pipe 31, a gas discharge main pipe 32, a gas conveying branch pipe 33, a gas discharge branch pipe 34 and a gas pump, wherein one end of the gas conveying branch pipe is embedded into the clapboards, and the other end of the gas conveying branch pipe is connected with the main pipe; the sealed container 2 comprises a bearing plate 21, a sealing plate 22, an internal bearing bowl 23 and an upper cover 24, wherein two air ducts 25 are arranged on the bearing plate 21, and the two air ducts 25 are respectively communicated with a gas conveying branch pipe 33 and a gas discharging branch pipe 34.

Further, a gas delivery branch pipe 33 and a gas discharge branch pipe 34 are embedded inside the carrying plate 21, and one end portion of each of the gas delivery branch pipe 33 and the gas discharge branch pipe 34 is embedded inside the partition plate 11, and extends upward from the upper surface of the partition plate 11 from the inside of the partition plate 11 through the surface of the partition plate 11 via a gas delivery nozzle 36 and a gas discharge nozzle 37.

Further, a gas flow meter and a solenoid valve are provided at the junction of the gas delivery manifold 31/gas discharge manifold 32 and the gas delivery branch pipe 33/gas discharge branch pipe 34.

Further, the gas flow meter and the electromagnetic valve are connected with the controller 13, so that the opening and closing of the gas path and the control of the gas flow are realized.

Further, the gas inlet end of the gas delivery main pipe 31 is connected with a gas pump for inputting gas, and the gas outlet end of the gas discharge main pipe 32 is connected with a gas pump for pumping gas; the gas pump for feeding gas is connected to a nitrogen gas cylinder 35.

Further, the gas pumps for inputting gas, the gas delivery manifold 31, the gas discharge manifold 32, the gas delivery branch pipes 33, the gas discharge branch pipes 34, and the gas pump systems for pumping gas are arranged in two symmetrical groups.

Further, the carrier plate 21 is a circular plate having a concave shape in a longitudinal section, an internal thread is provided on an inner side wall of a protrusion at both sides of the concave shape, and the upper cover 24 is a bell-shaped member having an opening at a lower portion. The lower part of the side wall of the box body is provided with an external thread.

Further, the upper part of the air duct 25 is cylindrical and deep upwards, the lower part is a through hole which is matched with the upper part in shape and penetrates through the bearing plate 21, and the upper part and the lower part form a complete and through air duct 25.

The autologous fat particle cryopreservation method of the autologous fat particle cryopreservation equipment comprises the following steps:

firstly, loading the fat particles after purification, cleaning and filtration into a sealed container 2;

thirdly, the sealed container 2 filled with the fat particles is placed into the low-temperature preservation equipment provided by the application, and the freezing and cooling are started;

fourthly, in the initial stage of freezing and cooling, the internal space of the sealed container 2 is cleaned by gas;

a fifth step of filling the sealed container 2 with gas before the fat has completely entered the frozen state (i.e. the fat itself has reached a predetermined freezing temperature of-60 to-80) after the gas washing step;

sixth, after the fat is completely frozen, the gas pump for gas input and the gas pump for gas extraction are turned off by the controller 13, and the electromagnetic valves located in the gas delivery manifold 33 and the gas discharge manifold 34 are turned off, so that the atmosphere inside the hermetic container 2 is stabilized, and a stable atmosphere frozen state is entered.

Further, step six is specifically that after the fat is completely frozen, the input gas pump and the electromagnetic valve located on the gas delivery branch pipe 33 are firstly closed, the metering is performed through the gas flow meter located on the gas discharge branch pipe 34, 30% of the volume of the gas in the internal volume of the sealed container 2 is pumped out through the gas pump for pumping out the gas and the gas discharge branch pipe 34, and the pumping-out gas pump and the electromagnetic valve located on the gas discharge branch pipe 34 are closed, so that the atmosphere in the sealed container 2 is stabilized, and the frozen state of the stable atmosphere is achieved.

The low-temperature preservation method of autologous fat particles provided by the invention can avoid fat pollution and improve the survival rate of later-stage fat transplantation.

Drawings

Fig. 1 is a schematic structural diagram of the cryopreservation of autologous fat granules provided by the invention.

Fig. 2 is a schematic structural diagram of the autologous fat granule cryopreservation sealed container 2 provided by the invention.

Detailed Description

The present invention will now be described in more detail with reference to the accompanying drawings, in which preferred embodiments of the invention are shown, it being understood that one skilled in the art may modify the invention herein described while still achieving the beneficial results of the present invention. Accordingly, the following description should be construed as broadly as possible to those skilled in the art and not as limiting the invention.

In the interest of clarity, not all features of an actual implementation are described. In the following description, well-known functions or constructions are not described in detail since they would obscure the invention in unnecessary detail. It will of course be appreciated that in the development of any such actual embodiment, numerous implementation-specific details must be set forth in order to achieve the developer's specific goals.

In order to make the objects and features of the present invention more comprehensible, embodiments of the present invention are described in detail below with reference to the accompanying drawings. It is to be noted that the drawings are in a very simplified form and are intended to use non-precision ratios for the purpose of facilitating and clearly facilitating the description of the embodiments of the invention.

The low-temperature preservation method of the autologous fat particles is based on the following low-temperature preservation equipment of the autologous fat particles. As shown in fig. 1, the apparatus has a refrigerated cabinet 1 body, a partition 11 located in the refrigerated cabinet 1 body, an air passage system, a sealed container 2 and a controller 13.

The body of the refrigerated cabinet 1 comprises two parts, an accommodating space at the upper part and an air supply cavity at the lower part. The refrigerator 1 body is provided with necessary refrigeration equipment, a sealing cabinet door and heat insulation parts to ensure the refrigeration effect. The accommodating space is used for accommodating the sealed container 2 for storing the frozen fat particles, and the accommodating space is divided into a plurality of, for example, 4, 6 or 8 spaces by a plurality of fixedly installed partition plates 11.

The gas path system comprises a gas delivery main pipe 31, a gas discharge main pipe 32, a gas delivery branch pipe 33, a gas discharge branch pipe 34 and a gas pump, wherein one end of the gas delivery branch pipe is embedded in the partition plate, and the other end of the gas delivery branch pipe is connected with the main pipe. Wherein the gas delivery branch pipe 33 and the gas discharge branch pipe 34 are embedded inside the partition plate, one end portion of each of the gas delivery branch pipe 33 and the gas discharge branch pipe 34 is embedded inside the partition plate 11, and the gas delivery nozzle 36 and the gas discharge nozzle 37 penetrate the surface of the partition plate 11 from inside the partition plate 11 to extend upward from the upper surface of the partition plate 11, thereby achieving delivery and discharge of the gas into the sealed container 2 in which the frozen fat particles are placed; the other end portions of the gas delivery branch pipes 33 and the gas discharge branch pipes 34 are connected to the gas delivery header pipe 31 and the gas discharge header pipe 32, respectively, so that the gas passages are connected.

And a gas flow meter and an electromagnetic valve are arranged at the joint of the gas conveying main pipe 31/the gas discharge main pipe 32 and the gas conveying branch pipe 33/the gas discharge branch pipe 34, and the gas flow meter and the electromagnetic valve are connected with the controller 13, so that the opening and closing of the gas path and the control of the gas flow are realized. The gas inlet end of the gas delivery manifold 31 is connected with a gas pump for inputting gas, and the gas outlet end of the gas discharge manifold 32 is connected with a gas pump for pumping gas. The gas pump for feeding gas is connected to a nitrogen gas cylinder 35. The gas pump for supplying gas is capable of introducing clean nitrogen gas into the interior of the apparatus, for example, the interior of the hermetic container 2, through the gas delivery manifold 31, the gas delivery manifold 33 and the gas delivery nozzle 36, under the control of the controller 13.

Preferably, for better control of the gas flow, the gas pump for gas input, the gas delivery manifold 31, the gas discharge manifold 32, the gas delivery branch 33, the gas discharge branch 34, and the gas pump system for gas extraction may be arranged in two symmetrical groups (as shown in fig. 1). Of course, if the volume of the cabinet 1 itself is small, the above-described system may be provided as a single unit for the sake of simplifying the equipment or reducing the cost.

In the individual storage spaces, which are divided by the partitions 11, in the interior of the refrigerated cabinet 1, a sealed container 2 is placed in each case in a separate, individual fit, the sealed containers 2 containing the fat particles to be frozen. As shown in FIG. 2, the sealed container 2 comprises a bearing plate 21, a sealing plate 22, an inner bearing bowl 23 and an upper cover 24. The bearing plate 21 is a circular plate with a concave longitudinal section, and internal threads are arranged on the inner side walls of the protrusions on the two sides of the concave longitudinal section, so that the bearing plate is convenient to connect with other components. The carrier plate 21 is made of stainless steel. Two hollow cylindrical air ducts 25 are provided on the bottom surface of the concave recess of the support plate 21, the upper part of the air duct 25 is cylindrical at the upper depth, the lower part is a through hole which is matched with the upper part in shape and penetrates through the support plate 21, and the upper part and the lower part form a complete and through air duct 25. The two air channels 25 correspond to the gas delivery nozzle 36 and the gas discharge nozzle 37 of the carrier plate 21 corresponding to the sealed container 2, respectively, and the gas delivery nozzle 36 and the gas discharge nozzle 37 can be inserted into the two air channels 25, respectively, so as to connect the sealed container 2 with the air channel system. The inner wall material of the air duct 25 is preferably a low temperature resistant rubber material, and the diameter size of the material is 2-3mm smaller than that of the gas delivery nozzle 36 and the gas discharge nozzle 37, so that the tight fit of the gas delivery nozzle 36 and the gas discharge nozzle 37 when inserted is realized.

A sealing plate 22 is arranged on the bottom surface of the concave part of the bearing plate 21, the sealing plate 22 is a circular flat plate and is made of low-temperature-resistant rubber material. The outer edge of the plate is dimensioned to fit and form a tight fit with the concave shape of the carrier plate 21. Two through holes are also provided in the sealing plate 22 at positions corresponding to the air passages 25 to facilitate passage and extension of the air passages 25. An inner bearing bowl 23 is fixedly attached to the upper surface of the sealing plate 22 and serves to contain the fat particles. On the upper surface of the sealing plate 22 and outside the inner carrying bowl 23, there is also mounted an upper cover 24. The upper cover 24 is a bell-shaped member having a lower opening. The lower part of the side wall of the baffle plate is provided with an external thread which is convenient to be connected with the baffle plate 11.

During the use process, the bearing plate 21 is firstly inserted into the gas delivery nozzle 36 and the gas discharge nozzle 37 through the air duct 25, so that the bearing plate 21 is fixed on the partition plate 11, then the sealing plate 22 fixedly connected with the inner bearing bowl 23 is arranged on the concave surface of the bearing plate 21, and finally the upper cover 24, the bearing plate 21 and the sealing plate 22 are pressed and arranged in a matching manner through the threaded matching of the upper cover 24 and the bearing plate 21, so that the sealed closing of the sealed container 2 is realized.

The air path system with the partition plate 11 and the part embedded into the partition plate 11 can effectively separate the space, realize the independent control of the atmosphere inside each sealed container 2, and effectively ensure the accurate control of the freezing condition.

The method for cryopreserving autologous fat particles provided by the present invention based on the above-mentioned cryopreservation apparatus is described in detail below. The method for preserving the autologous fat particles at the low temperature comprises the following steps:

in the first step, the fat particles after purification, washing and filtration are loaded into the sealed container 2.

Specifically, to purify, wash the fat particle after filtering, put into inside and bear the bowl 23, insert the loading board 21 with gas delivery mouth 36 and gas emission mouth 37 respectively through air duct 25 and close, make loading board 21 be fixed in on baffle 11, install the closing plate 22 of the inside bearing bowl 23 of fixed connection on the sunken surface of loading board 21 afterwards, through the screw-thread fit with upper cover 24 and loading board 21, compress tightly upper cover 24, loading board 21 and closing plate 22 installation cooperation, thereby realized the sealed loading of fat particle.

Preferably, the fat particles may be provided with a cryoprotectant, which may be dimethyl sulfoxide and trehalose, or other cryoprotectants commonly used in the art.

In a second step, the sealed container 2 containing the fat particles is pre-cooled.

Specifically, the sealed container 2 loaded with fat particles is placed in a refrigerator or other refrigeration equipment commonly used in the art for primary cooling. So that the temperature of the sealed container 2 and the fat particles drops to around 0 degrees celsius.

Preferably, the pre-cooling time is determined according to the volume of the fat particles, and the pre-cooling time is 15 minutes per 5ml of fat.

And thirdly, placing the sealed container 2 filled with the fat particles into the low-temperature preservation equipment provided by the application, and starting freezing and cooling.

Specifically, the gas delivery nozzle 36 and the gas discharge nozzle 37 are inserted into the two air passages 25 of the hermetic container 2, respectively. The connection between the accommodating space in the sealed container 2 and the air path system is realized. Then the sealing door of the refrigerated cabinet 1 is closed, and the refrigeration temperature is started to be reduced, wherein the refrigeration temperature is-60 to-80 ℃.

And fourthly, in the initial stage of freezing and cooling, carrying out gas cleaning on the inner space of the sealed container 2.

The atmosphere in the internal space of the sealed container 2 is gas-purged in synchronization with the time of starting the freezing and cooling. The method comprises the following specific steps: the controller 13 controls the air pump for inputting air and the air pump for pumping air to work, the air pump for pumping air pumps original air in the sealed container 2 out through the air discharge nozzle 37, and the air pump for inputting air pumps pure and dry nitrogen into the sealed container 2 through the air delivery nozzle 36. Through the flowing cleaning of the gas, the atmosphere in the sealed container 2 is purified, particle pollution in the original gas can be removed, the higher oxygen partial pressure in the sealed container 2 can be reduced, and the fat preservation effect is improved.

Preferably, during this gas purging step, the controller 13 controls the flow rate of the gas flow by controlling gas flow meters located on the gas delivery manifold 33 and the gas discharge manifold 34. During the cleaning step, the gas flow rate in the gas delivery manifold 33 does not exceed the gas flow rate in the gas discharge manifold 34, thereby further facilitating the sufficient discharge of the gas and particles inside the hermetic container 2. Further preferably, the time of the above-mentioned washing step is not more than 10 minutes, preferably 5 minutes, in order not to affect the freezing and cooling effect, since the gas flow and the relatively low gas pressure affect the heat conduction. And ensures that the gas in the internal volume of the sealed container 2 is displaced a minimum of one time, preferably 2-3 times, during the cleaning step. Therefore, the necessary conditions can be achieved by selecting an appropriate gas flow rate according to the volume of the sealed container 2.

Fifth, after the gas purging step, the sealed container 2 is filled with gas before the fat has completely entered its frozen state (i.e. the fat itself has reached a predetermined freezing temperature of-60 to-80).

After the gas purging step, the gas flow rate in the gas delivery manifold 33 is made greater than the gas flow rate in the gas discharge manifold 34 by controlling the gas flow meter by the controller 13 before the fat is completely frozen (i.e., the fat itself reaches a predetermined freezing temperature of-60 to-80 degrees celsius), the gas flow rate in the gas discharge manifold 34 not exceeding 40 sccm. Preferably, the gas flow on the gas delivery manifold 33 is 10-20sccm greater than the gas flow on the gas discharge manifold 34. In addition, in the step of gas filling, under the conditions of lower gas flow rate and higher nitrogen gas pressure, the gas cleaning can be realized, the quick heat transfer can be realized, the fat cooling efficiency is improved, the quick cooling of fat is realized, and the fat quality is ensured.

Furthermore, the judgment of the period of time before the fat has completely entered the frozen state after the gas washing step (i.e. the fat itself has reached a predetermined freezing temperature of-60 to-80) can be calculated from the volume of fat, which has been completely entered the frozen state after a freezing time of 20-30 minutes for every 5ml of fat according to experimental data analysis.

Alternatively, in the sixth step, after the fat is completely frozen, the input gas pump and the electromagnetic valve located on the gas delivery branch pipe 33 are first closed, the gas is measured by the gas flow meter located on the gas discharge branch pipe 34, and the gas inside the sealed container 2 is pumped out by the gas pump for pumping out the gas and the gas discharge branch pipe 34, wherein the flow rate and the time of the gas pumping out are controlled as follows: depending on the internal volume of the sealed container 2, 30% by volume of the internal volume is evacuated. Subsequently, the suction gas pump and the electromagnetic valve provided in the gas discharge branch pipe 34 are closed, so that the atmosphere inside the hermetic container 2 is stabilized, and a stable atmosphere frozen state is entered. Thus, in a stable atmosphere frozen state, the air pressure is lower, and the influence of excessive temperature conduction and atmosphere on the activity of fat is avoided.

The freshly removed fat was washed and filtered and then split into two portions, one portion was frozen for two weeks by the preservation method provided herein (sample 1), the other portion was frozen by the conventional cryopreservation method (sample 2) after adding the same cryoprotectant and then placing in a-80 ℃ freezer for two weeks), and after two weeks, the

above samples

1, 2 were subjected to the same thawing recovery treatment and tested. The glucose transport amount of sample 1 was 1.575. + -. 0.075mmol/L before freezing, 1.524. + -. 0.052mmol/L after freezing, the glucose transport amount of sample 2 was 1.594. + -. 0.073mmol/L before freezing, and 1.248. + -. 0.063mmol/L after freezing. The

above samples

1 and 2 were each subjected to a biopsy of an animal, and the volume of fat of the

samples

1 and 2 was recorded to be 1ml before the transplantation, and two weeks after the transplantation, the sample was taken from the transplantation site. First, the graft is found at the graft site, the skin muscle is cut 0.3cm around the periphery of the graft, the fat graft is removed, the surrounding tissue and the peripheral envelope are detached and repaired to fully expose the pure fat graft, measured by tissue observation and by a fine gauge micro-scale tube. Significant inflammatory responses were observed in both the graft and surrounding tissue of

samples

1, 2, with some adipose tissue and abscesses visible within the fat envelope, and sample 2 was more severe than sample 1. Through the measurement of a micro-scale test tube of a precision measurer, the fat volume of the sample 1 is 0.628 +/-0.035 ml, and the fat volume of the sample 2 is 0.521 +/-0.014 ml, so that the fat transplantation group preserved by the freezing method provided by the application is superior to the conventional fat transplantation group preserved by freezing, and part of cells preserved by the freezing method provided by the application have better functions and forms and higher survival rate.

By the method for preserving the autologous fat particles at the low temperature, fat pollution can be avoided, and the survival rate of later fat transplantation is improved.

The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.

Claims

1. A method for preserving autologous fat particles at low temperature is characterized in that: the autologous fat particle low-temperature preservation method uses autologous fat particle low-temperature preservation equipment, and the autologous fat particle low-temperature preservation equipment comprises:

the refrigerator comprises a refrigerator body, a partition plate positioned in the refrigerator body, a gas path system, a sealed container and a controller;

the refrigerator body comprises two parts, namely an accommodating space positioned at the upper part and an air supply cavity positioned at the lower part; the storage space is used for storing a sealed container for storing frozen fat particles, and the storage space is divided into a plurality of storage spaces by a plurality of fixedly installed partition plates;

the gas path system comprises a gas conveying main pipe, a gas discharging main pipe, a gas conveying branch pipe, a gas discharging branch pipe and a gas pump, wherein one end of the gas conveying branch pipe is embedded in the partition plate, and the other end of the gas conveying branch pipe is connected with the main pipe; wherein the gas delivery manifold and the gas discharge manifold are embedded inside the partition, one end of each of the gas delivery manifold and the gas discharge manifold is embedded inside the partition, and the gas delivery nozzle and the gas discharge nozzle penetrate the surface of the partition from the inside of the partition and extend upward from the upper surface of the partition, thereby achieving delivery and discharge of the gas into the sealed container in which the frozen fat particles are placed; the other end parts of the gas conveying branch pipe and the gas discharge branch pipe are respectively connected with a gas conveying main pipe and a gas discharge main pipe, so that the connection of gas circuits is realized;

a gas flowmeter and an electromagnetic valve are also arranged at the joint of the gas conveying main pipe and the gas discharge main pipe and the joint of the gas conveying branch pipe and the gas discharge branch pipe, and the gas flowmeter and the electromagnetic valve are connected with a controller; the gas inlet end of the gas delivery header pipe is connected with a gas pump for inputting gas, and the gas outlet end of the gas discharge header pipe is connected with a gas pump for pumping gas; the air pump for inputting air is connected with a nitrogen gas bottle; through the control of the controller, the gas pump for inputting gas can lead clean nitrogen into the equipment through the gas delivery main pipe, the gas delivery branch pipe and the gas delivery nozzle;

in the single storage space divided by the partition board in the refrigerated cabinet, a sealed container is respectively and independently arranged in a matched manner, and the sealed container stores fat particles to be frozen; the sealed container comprises a bearing plate, a sealing plate, an internal bearing bowl and an upper cover; the bearing plate is a circular plate with a concave longitudinal section, and internal threads are arranged on the inner side walls of the protrusions on the two sides of the concave shape, so that the bearing plate is convenient to connect with other components; the bearing plate is made of stainless steel material; two hollow cylindrical air ducts are arranged on the bottom surface of the concave depression of the bearing plate, the upper part of each air duct is in a cylindrical shape at the upper depth, the lower part of each air duct is a through hole which is matched with the shape of the upper part and penetrates through the bearing plate, and the upper part and the lower part form a complete and through air duct; the positions of the two air channels respectively correspond to the gas conveying nozzle and the gas discharging nozzle of the bearing plate corresponding to the sealed container, and the gas conveying nozzle and the gas discharging nozzle can be respectively inserted into the two air channels, so that the sealed container is connected with the gas circuit system;

the bottom surface of the concave recess of the bearing plate is provided with a sealing plate, and the position of the sealing plate corresponding to the air channel is also provided with two through holes, so that the air channel can conveniently pass through and extend out; an inner bearing bowl is fixedly connected to the upper surface of the sealing plate and used for containing fat particles; an upper cover is also arranged on the upper surface of the sealing plate and outside the inner bearing bowl; the upper cover is a bell-shaped component with a lower opening; the lower part of the side wall of the baffle plate is provided with external threads, so that the baffle plate can be conveniently connected with the baffle plate;

the method for preserving the autologous fat particles at the low temperature comprises the following steps:

firstly, loading the fat particles after purification, cleaning and filtration into a sealed container,

specifically, the purified, cleaned and filtered fat particles are placed into an internal bearing bowl, a bearing plate is respectively inserted into a gas conveying nozzle and a gas discharging nozzle through an air duct 25, so that the bearing plate is fixed on a partition plate, then a sealing plate fixedly connected with the internal bearing bowl is arranged on the concave surface of the bearing plate, and finally the upper cover, the bearing plate and the sealing plate are tightly pressed, installed and matched through the threaded matching of the upper cover and the bearing plate, so that the sealed loading of the fat particles is realized;

secondly, pre-cooling the sealed container filled with the fat particles;

specifically, the sealed container loaded with the fat particles is subjected to primary cooling, so that the temperature of the sealed container and the fat particles is reduced to be about 0 ℃;

thirdly, the sealed container filled with the fat particles is placed into the low-temperature preservation equipment for the autologous fat particles, and the freezing and the temperature reduction are started,

specifically, a gas delivery nozzle and a gas discharge nozzle are respectively inserted into two air ducts of a sealed container to realize the connection between an accommodating space inside the sealed container and a gas path system, then a sealing door of a refrigerator body is closed, and freezing and cooling are started, wherein the freezing temperature is-60 to-80 ℃;

fourthly, in the initial stage of freezing and cooling, the internal space of the sealed container is cleaned by gas,

the method comprises the following specific steps: the controller controls the air pump for inputting air and the air pump for pumping air to work, the air pump for pumping air pumps the original air in the sealed container through the air discharge nozzle, and the air pump for inputting air pumps pure and dry nitrogen into the sealed container through the air delivery nozzle;

during the gas cleaning step, the controller controls the flow rate of the gas flow by controlling gas flow meters on the gas conveying branch pipe and the gas discharge branch pipe, during the cleaning step, the gas flow rate on the gas conveying branch pipe does not exceed the gas flow rate on the gas discharge branch pipe, the time of the cleaning step does not exceed 10 minutes, and the gas in the internal volume of the sealed container is replaced at least once within the time of the cleaning step;

a fifth step of filling the sealed container with gas before the fat is completely put into a frozen state at a predetermined freezing temperature of-60 to-80 after the gas purging step,

the method comprises the following specific steps: after the gas cleaning step and before the fat completely enters a freezing state, controlling a gas flow meter by a controller to realize that the gas flow on the gas conveying branch pipe is greater than the gas flow on the gas discharge branch pipe, wherein the gas flow on the gas discharge branch pipe is not more than 40 sccm;

sixthly, after the fat completely enters the freezing state, closing the air pump for inputting air and the air pump for pumping out air through the controller, and closing the electromagnetic valves positioned on the air conveying branch pipe and the air discharging branch pipe to ensure that the atmosphere in the sealed container is stable and enters a stable atmosphere freezing state; or firstly closing the input gas pump and the electromagnetic valve on the gas delivery branch pipe, metering by a gas flow meter on the gas discharge branch pipe, and pumping gas in the sealed container by the gas pump for pumping gas and the gas discharge branch pipe, wherein the flow rate and time of gas pumping are controlled as follows: gas of 30% volume of the inner volume is pumped out according to the inner volume of the hermetic container, and then, the pump for pumping out gas and the solenoid valve located on the gas discharge branch pipe are closed, so that the atmosphere inside the hermetic container is stabilized to enter a stable atmosphere frozen state.