JP5476556B2 - Cell transport container - Google Patents

Description

  The present invention relates to a cell transport container for transporting a biological sample containing, for example, a regenerated tissue manufactured in a cell processing facility, while maintaining a culture environment related to temperature, pressure, cleanliness, and the like.

  Regenerative medicine using a biological sample such as a regenerated tissue produced by culturing a small amount of cells to restore the function of a lost organ or the like is a technique that is expected to be effective for diseases for which there has been no conventional treatment. The manufacturing process of biological samples such as regenerative tissues used in regenerative medicine is based on Good Manufacturing Practice (GMP), which is a standard for manufacturing control and quality control of pharmaceuticals. Manufacture is performed in a cell processing center (CPC) and follows a standard operating procedure (SOP) that satisfies GMP. In Japan, the laws and regulations set by the Ministry of Health, Labor and Welfare have already been enforced in GMP (for example, Ministry of Health, Labor and Welfare Order No. 179, Drug Development No. 480). Outside Japan, relevant laws and regulations are being enforced mainly by Western institutions (eg, US Food and Drug Administration, European Commission).

  A biological sample such as a regenerated tissue is manufactured by a manufacturing worker who has specialized cell culture technology in an environment where safety and cleanliness are controlled as defined by these regulations. Therefore, a great labor cost, labor, and operation cost are generated. At present, since almost all manufacturing processes are performed manually, there is a limit to the increase in production volume. As a result, the manufacturing cost is high, which hinders the spread of regenerative medical treatment. Therefore, at the initial stage of practical application of regenerative medicine, it is assumed that the products will be manufactured at a small number of CPCs as production bases, shipped to medical institutions in various places, that is, transported and used for treatment. In addition, the cell processing facility for manufacturing and the medical institution for performing treatment are separated from each other through a daily space that is less clean than the culture environment, even if they are in the same site. Therefore, although the distance varies, the work of transport always occurs.

  In general, transportation of biological samples such as regenerative tissues is routinely performed in research and development. However, there are various transport methods, and they are appropriately selected according to the cell type and use after transport. A standardized transportation method has not been established. For example, when transporting at a constant temperature that is the same as the culture temperature (for example, about 37 ° C), when transporting in the open air without temperature control (for example, 10 to 37 ° C), the cells are kept in a refrigerated state. It is used when it is transported while suppressing its metabolism (for example, 4 ° C.) and when it is transported in a frozen state (for example, −20 ° C. in liquid nitrogen). Unlike the manufacturing process, the transportation process is affected by temperature, pressure, shock, vibration, and the like. In addition, the environment during transportation is air-conditioning controlled by a HEPA filter (High Efficiency Particulate Air Filter), and the cleanliness is much lower than in a culture environment in which organisms such as bacteria and particles are hardly present. In order to use transported regenerative tissue in clinical practice, these effects must be controlled and the regenerated tissue must meet the quality required by GMP even after transport.

  The external environment that passes during transportation first has a different temperature compared to the culture environment. In the culture environment of the cell treatment facility, the temperature is appropriately selected according to the cell type and application. In many cases, the temperature during cultivation is 37 ° C. Since the thermostatic bath is used for temperature control, the temperature change width is small. On the other hand, during transportation, the temperature of the external environment varies depending on the location. Therefore, in order to eliminate the influence of the temperature of the external environment, a mechanism for keeping the temperature inside the cell transport container constant is necessary. Moreover, in order to reduce the influence of the temperature difference with the external environment, it is necessary to install a heat insulating material or the like around the cell transport container to ensure heat insulation. Furthermore, it is desirable that the time for keeping the internal temperature constant is sufficiently long. In the future, it is assumed that there will be a situation where the aircraft is transported to a distant place such as overseas, or a waiting time will occur in preparation for examination or surgery after arriving at the medical institution that is the destination of transportation.

  Pressure is another factor that differs between the external environment during transport and the culture. Although the culture environment is normal pressure of about 1 atm, when transporting, especially when using an aircraft, the pressure in the cabin is around 0.8 atm. When a general culture vessel is exposed to a reduced pressure, the gas dissolved in the medium is eluted and the pH changes, which makes it unsuitable for cell growth. In addition, the culture medium may leak from the gap between the lid of the culture container and the main body. In that case, cleanliness is lost and biological contamination occurs. In order to avoid these, an airtight mechanism is necessary to eliminate the influence of pressure. Airtightness can be achieved by means such as screws and hinged metal fittings. However, as the volume of the airtight object increases, the necessary parts increase and the weight of the entire cell transport container increases. As a result, the burden on transportation workers increases.

The cleanliness of other factors that differ between transport and culture is defined by laws and regulations. For example, according to European GMP, the place where cell culture facilities are cultured (hereinafter referred to as the culture area) is Grade B (the number of suspended bacteria is 10 CFU / m ³ or less, the diameter is 0.5 μm or more and the number of particles is 350,000 / m ³ or less). Sex is required. In the safety cabinet installed in the culture area, the cells are directly touched through the medium, such as cell purification and medium exchange. However, the cleanliness is Grade A (the number of suspended bacteria is 1 CFU / m ³ or less, the diameter is 0.5 μm or more and 3500 particles / m ³ or less) are required. In order to maintain cleanliness, air circulation through HEPA filters, control of pressure difference between rooms, flow management of workers and goods, and daily cleaning work are performed. By the way, the temperature of the biological sample handled in the culture area is controlled by a thermostat and a heat block or the like installed in the safety cabinet as necessary. On the other hand, the temperature in other places is not a value suitable for culture, and is generally about 25 ° C. Therefore, when transporting a biological sample, it is necessary to bring a cell transport container having a mechanism for maintaining the temperature to the vicinity of the safety cabinet. That is, in the safety cabinet, the cell transport container packed in a transportable shape is quickly accommodated in the cell transport container while maintaining the temperature of the culture container with a heat block or the like. This makes it possible to transport without being affected by temperature changes. For this purpose, the cell transport container needs to have cleanliness that can be brought into a culture area of grade B.

  There are several reports on the transport technology of biological samples such as regenerative tissues as conventional technology against this background. First, regarding a temperature maintenance technique, there is one that uses a heat panel or the like using a heat insulating material and electric power. This always has the possibility of failure in wiring or the like, and lacks certainty. In addition, there is a possibility that radio waves may be emitted during operation, and in that case, there is a possibility that an influence will be exerted on the equipment of a medical institution.

  On the other hand, a temperature maintenance technique that does not use electric power has also been reported. Document 1 uses a heat insulating material and a heating device using an oxidation action by a chemical catalyst. Although electric power is not used, it is difficult to control the amount of heat generated by a chemical reaction, and it is difficult to keep the internal temperature constant. Further, since the chemical reaction occurs irreversibly, the heating device cannot be used repeatedly. Cost is high.

  Documents 2 and 3 use a heat storage material such as hydrocarbon which is a pure substance. When the molten heat storage material solidifies, the fact that the temperature becomes constant at the melting point is utilized. In the case of this method, since the melting point is a value inherent to the substance, the certainty is higher than the method using electric power.

  The temperature maintenance performance also depends on the heat insulation performance and the heat insulation efficiency. Documents 1, 2, and 3 all have a single heat insulating material. Document 1 houses a biological sample surrounded by a heating device and a buffer material inside a heat insulating material. The heating device is placed next to the cushioning material. Therefore, when warming the interior, the cushioning material that does not need to be warmed is also warmed at the same time. This is poor heat retention efficiency. The same applies to Documents 2 and 3, and the heat storage efficiency is poor even with respect to the cushioning material disposed inside the heat insulating material. By the way, it is desirable that the temperature maintenance performance required for the cell transport container is sufficiently long. In the future, it is expected to be transported overseas. In addition, it is assumed that in the medical institution after arrival, preparation for treatment is performed while the culture container is accommodated in the cell transport container. To maintain the temperature for a sufficiently long time, it is possible to increase the amount of chemicals and heat storage materials, but the total weight of the cell transport container also increases. As a result, the burden on transportation workers increases. In order to avoid this, it is necessary to improve heat insulation performance and heat retention efficiency.

  With regard to pressure, airtightness is necessary, especially when transported by aircraft. For example, in the case of Documents 1, 2, and 3, airtightness can be achieved by installing a metal fitting or the like on the outermost side of the container and firmly closing the lid. However, since the volume of the airtight object is large, the necessary parts become large and the overall weight becomes large. The burden on transportation workers increases.

  Regarding cleanliness, as described above, it is necessary to carry the cell transport container to the side of the safety cabinet in the culture area. For example, in the case of Documents 1, 2, and 3, it is difficult to ensure the cleanliness of the cell transport container. Although it is necessary to sterilize the cell transport container in advance, Documents 1, 2, and 3 do not use materials that can be sterilized. Even if a material that can be sterilized is changed and sterilization becomes possible, the equipment (for example, an autoclave device) required to sterilize the entire cell transport container is required to be huge. Further, even if sterilization is performed, in order to bring the cell transport container to the culture area, it is necessary to pass through a pass box that connects the rooms in the CPC. In general, the size of a pass box is a small space of about 50 cm square. It is necessary to make the size of the cell transport container so that it can pass through the pass box or to increase the size of the pass box. In other words, the cell transport container needs to be clean enough to be brought into the culture area, and to be able to pass through the pass box.

JP 2004-217290 A JP 2007-191209 A JP 2006-232331 A

  As described above, a cell transport container for transporting a biological sample such as a regenerated tissue needs a function of maintaining a culture environment. During transportation, it is affected by temperature, pressure, shock, and vibration, unlike the culture environment at the time of manufacture. Also, the cleanliness is greatly different. In order for a biological sample such as a regenerated tissue to maintain a good state after transportation, it is necessary to eliminate these effects. That is, there is a need for a technique for maintaining the temperature constant throughout the entire transportation process and a technique for eliminating the effects of pressure changes when transporting by aircraft or the like. Moreover, since it is necessary to carry a cell transport container to the clean culture area in a cell processing facility, the technique which ensures cleanliness is also required.

  The present invention is capable of transporting a biological sample such as a regenerated tissue while maintaining the same temperature and pressure as the culture environment of the cell treatment facility, and bringing it into the cell treatment facility while ensuring cleanliness. An object of the present invention is to provide a cell transport container capable of performing the above.

  In order to achieve the above object, the cell transport container of the present invention is arranged on at least a part of the inside of the first heat-insulating container, the first heat-insulating container having the entire wall surface constituted by a high-performance heat insulating material such as a vacuum heat insulating material. Low thermal conductivity cushioning material made of foamed polystyrene or urethane foam to absorb shocks and vibrations, and high performance thermal insulation materials such as vacuum thermal insulation material placed inside the low thermal conductivity cushioning material A second heat insulating container that constitutes the entire wall surface, an airtight container disposed inside the second heat insulating container, and a culture container storage unit disposed inside the airtight container are provided. Here, a 1st heat insulation container and a 2nd heat insulation container have an effect which reduces the transfer efficiency of the heat between a 1st heat insulation container and a 2nd heat insulation container by contacting only through buffer material / air. .

  Further, in the cell transport container of the present invention, an air bag in which air is enclosed in a bag made of a material such as vinyl chloride, together with the low thermal conductivity buffer material, between the first heat insulating container and the second heat insulating container. May be arranged. Thereby, it has the effect which buffers the vibration and impact which arise during transportation by the elasticity which an air bag has.

  In the cell transport container of the present invention, the first heat storage material box enclosed with the heat storage material is disposed inside the second heat insulation container and in close contact with the airtight container. May be. It has the effect that the temperature of the hermetic container is maintained by the release / absorption of the amount of heat by the heat storage material of the first heat storage material box.

  Further, the cell transport container of the present invention comprises a second heat storage material box enclosed with a heat storage material, arranged inside the airtight container and in close contact with the culture container storage portion. May be. The second heat storage material box has an effect of maintaining the temperature of the airtight container.

  In the cell transport container of the present invention, a hydrocarbon may be used as the heat storage material. Thereby, temperature can be maintained constant reliably.

  In the cell transport container of the present invention, the first heat storage material box or the second heat storage material box may be divided into a plurality of smaller divided heat storage material boxes. This facilitates the movement of heat inside the heat storage material, and has an effect of improving the efficiency of releasing heat from the heat storage material.

  Moreover, the cell transport container of this invention WHEREIN: At least one part of the surface which comprises the said some division | segmentation heat storage material box may be comprised with highly heat conductive materials, such as stainless steel. This has the effect of further improving the heat conduction efficiency.

  Further, in the cell transport container of the present invention, a plurality of heat storage material-containing bags are arranged in the first heat storage material box, the second heat storage material box, or the divided heat storage material box, and a high heat conductive metal mesh is provided between the heat storage material-containing bags. Or you may arrange | position a highly heat conductive thin film. Thereby, the effect that the movement of the heat in a 1st and 2nd heat storage material box becomes easy is acquired.

  Further, in the cell transport container of the present invention, the high-performance heat insulating material of the second heat insulating container includes a low thermal conductive buffer material made of foamed polystyrene or urethane foam, and a high thermal conductive metal mesh or a high thermal conductive metal thin film. A plurality of rectangular parallelepiped structures that are alternately stacked may be arranged in a random direction so that there is no gap. By adopting such a structure, it has a buffer function by a low thermal conductivity buffer material made of foamed polystyrene, urethane foam, etc., and a high thermal conductivity of a high thermal conductivity metal thin film or a high thermal conductivity metal mesh such as stainless steel. It becomes possible.

  Further, in the cell transport container of the present invention, the culture container storage section and the second heat storage material box in which the heat storage material is enclosed can be sterilized by autoclaving or the like, and between the rooms in the cell processing facility. This is a size capable of passing through the inside of the pass box. Thereby, it is possible to carry in the state which maintained sterility to the room which cultures in a cell treatment facility.

  In the cell transport container of the present invention, a first temperature / pressure sensor for recording changes in temperature and pressure in the airtight container may be installed inside the airtight container. Thereby, the temperature and pressure in the airtight container during transportation can be measured.

  In the cell transport container of the present invention, a second temperature / pressure sensor that measures and displays the temperature and pressure outside the airtight container may be installed outside the airtight container. Thereby, temperature and pressure can be monitored during transportation.

  According to the cell transport container according to the present invention, a biological sample such as a regenerated tissue can be transported while maintaining the same temperature and pressure as the culture environment of the cell treatment facility. In addition, since a part of the cell transport container can be sterilized, it can be carried into the cell processing facility while ensuring cleanliness.

The figure which shows one Example of this invention and shows the structure of the perpendicular direction of a cell transport container. The figure which shows one Example of this invention and shows the structure of the horizontal direction of a cell transport container. The figure which shows one Example of this invention and shows the structure which installed the air bag to the heat insulation structure of a cell transport container. The figure which shows one Example of this invention and shows the structure of the vertical direction of the cell transport container at the time of dividing | segmenting a thermal storage material box. The figure which shows one Example of this invention and shows the structure of the state which accommodated the culture container in the airtight container in a cell transport container. The figure which shows one Example of this invention and shows a structure at the time of installing a metal mesh in the inside of a thermal storage material box. The figure which shows one Example of this invention and shows the structure of a highly heat conductive heat insulating material.

  Hereinafter, the cell transport container according to the present invention will be described in detail with reference to the drawings.

  The basic components of the cell transport container will be described with reference to FIGS. The cell transport container is a low thermal conductivity material made of foamed polystyrene or urethane foam for absorbing shock and vibration inside the first heat insulating container 101, 201 whose entire wall surface is composed of high performance heat insulating material such as vacuum heat insulating material. In addition, second heat insulating containers 103 and 203 whose entire wall surfaces are made of high performance heat insulating material such as vacuum heat insulating material are disposed inside. The low thermal conductivity cushioning materials 102 and 202 are arranged between the high-performance heat insulating materials arranged twice, and only air exists in the portions where the low thermal conductivity cushioning materials 102 and 202 are not provided. Air is also a material with low thermal conductivity. As a result, the low thermal conductivity buffer materials 102 and 202 and the air layer between the double high performance heat insulating materials have both a buffering effect and a heat insulating effect. Further, as shown in FIG. 3, a plurality of air bags 301 in which air is sealed in a bag made of a material such as vinyl chloride may be installed in the air layer. The installed air bag has elasticity and can serve as a cushioning material. That is, the cushioning effect can be improved by installing the air bag 301.

Inside the second heat insulating containers 103 and 203, the airtight containers 104 and 204 and the first heat storage material boxes 105 and 205 are disposed so as to surround the periphery of the airtight container. These are in close contact, and the effect of releasing / absorbing the amount of heat by the heat storage material of the first heat storage material box is efficiently transmitted to the airtight container. The hermetic containers 104 and 204 are all wall surfaces made of a high thermal conductivity material such as stainless steel, and can be hermetically sealed by means such as a hinge type metal fitting or a screw. The first heat storage material boxes 105 and 205 are filled with a heat storage material such as hydrocarbon, and for example, a plastic material such as polycarbonate is used. As the heat storage material, for example, a saturated chain hydrocarbon having a general formula of C _n H _{2n + 2} is used. Although the melting point varies depending on the carbon number of C, the number of C in the cell transport container is determined according to the value of the internal temperature to be kept constant. For example, in the case of n-Eicosane represented by the chemical formula of C ₂₀ H ₄₂ , the melting point is 36.4 ° C. Therefore, when this substance is enclosed in the heat storage material box, the internal temperature of the cell transport container can be maintained at about 36 ° C.

  The first heat storage material boxes 105 and 205 and the second heat storage material box 108 described later may be divided into smaller heat storage material boxes. When the temperature decreases, the heat storage material solidifies from the peripheral portion. The solidified heat storage material has low thermal conductivity. Further, convection with the liquid heat storage material that is not solidified and remains in the heat storage material does not occur. Therefore, the heat quantity of the liquid heat storage material confined inside the solidified heat storage material is greatly reduced in the efficiency of release to the outside of the heat storage material box. Here, by dividing the heat storage material box into smaller ones, heat transfer is facilitated inside the heat storage material. Therefore, the heat release efficiency from the heat storage material is improved. FIG. 4 shows an example in which the first heat storage material box is divided into smaller divided heat storage material boxes 401. Furthermore, you may change a part or all of the surface which comprises a thermal storage material box into highly heat conductive materials, such as stainless steel. Thereby, the heat conduction efficiency is further improved. However, generally, a high thermal conductivity material such as stainless steel is heavier than a plastic material such as polycarbonate. Furthermore, the smaller the size of the divided heat storage material box, the better the heat conduction efficiency, but the number of parts increases, so the handling as a cell transport container becomes complicated. In addition, since the amount of material constituting the heat storage material surface increases, the weight of the entire cell transport container also increases. It is necessary to use an optimal heat storage material box after grasping the required temperature maintenance performance.

  The inside of the airtight containers 104 and 204 is a second heat storage in which a culture container housing portion 107 and 207 made of a high thermal conductivity material such as stainless steel for housing the culture containers 106 and 206 and a heat storage material such as hydrocarbon are enclosed. A material box 108 and first temperature / pressure sensors 109 and 209 for recording changes in temperature and pressure in the hermetic container are arranged. The culture vessel storage portions 107 and 207 are positioned so that the second heat storage material box 108 surrounds them. In addition, these are in close contact with each other, and the heat conduction efficiency is high. The temperature / pressure sensors 109 and 209 measure the temperature and pressure in the airtight container being transported. However, the first temperature / pressure sensors 109 and 209 are placed in an airtight container, and monitoring data cannot be confirmed from the outside of the cell transport container. After arrival at a medical institution or the like, the data is taken out from the cell transport container and the data is confirmed. Therefore, the second temperature / pressure sensor 110 is installed outside the airtight container so that the data monitored during transportation can be confirmed. The second temperature / pressure sensor 110 is installed on a heat insulating lid 111 in which a lid portion of the first and second heat insulating containers and a low thermal conductivity buffer material made of foamed polystyrene, urethane foam or the like are integrated. A temperature sensor unit 113 installed between the first heat storage materials 105 and 205 and the hermetic containers 104 and 204 is connected to the second temperature / pressure sensor 110 via a cord 112. Thereby, the temperature between a 1st heat storage material and an airtight container is measured. For pressure, measure the pressure outside the hermetic vessel. A lid 114 for restraining the heat insulating lid 111 is installed on the upper side of the heat insulating lid 111. This keeps the contents from coming out during transport. A transparent window 115 is installed at the center of the lid 114. From this window, the temperature and pressure data that is always displayed by the second temperature / pressure sensor 110 installed inside the window can be confirmed.

  FIG. 5 shows a state in which various culture containers containing biological samples including regenerated tissue are accommodated in an airtight container. FIG. 5A shows a state in which the 6-well plates 501 are stacked in the vertical direction. FIG. 5B shows a state in which a 1.5 ml sample tube 502 is contained. FIG. 5C shows a state in which a 50 ml sample tube 503 is contained. Considering the shape of the culture vessel to be used and the state in which the biological sample is contained, the shape of the culture vessel storage portion is determined so as to be held at an optimal position during transportation. And what is necessary is just to arrange | position a thermal storage material box so that it may be enclosed.

  FIG. 6 shows a configuration for improving the heat conduction efficiency when the heat storage material is housed in a bag of vinyl chloride or the like with respect to the heat storage material put in the heat storage material box. The heat storage material may be directly enclosed in the heat storage material box and completely enclosed so as not to leak to the outside. However, it may be packed in a bag of vinyl chloride or the like and packed in a heat storage material box. In the latter case, there is an advantage that even if a heat storage material box having a different shape is used, it is only necessary to replace the packaged heat storage material. In the case where such a situation is assumed, in FIG. 6, a high thermal conductive metal mesh 603 such as stainless steel is disposed between the heat storage material-containing bags 602 packed in the heat storage material box 601. Thereby, it becomes possible for heat to move efficiently inside the heat storage material box. The high thermal conductive metal mesh 603 may be another component as long as it is light and has high thermal conductivity. For example, a metal thin film such as stainless steel may be used.

  FIG. 7 shows a high thermal conductivity buffer material packed in a high-performance heat insulating material constituting the second heat insulating container. FIG. 7A shows a basic unit constituting the high thermal conductivity buffer material. A low thermal conductivity buffer material 701 made of foamed polystyrene, urethane foam, or the like and a high thermal conductivity metal mesh 702 such as stainless steel are alternately laminated to form a rectangular parallelepiped shape. For example, a metal thin film such as stainless steel may be used instead of the high thermal conductive metal mesh such as stainless steel. Using this basic unit, as shown in FIG. 7 (B), a material in which directions are random and integrated so as not to have a gap by means of an adhesive or the like is used as a high thermal conductivity buffer material. A low thermal conductivity buffer material made of foamed polystyrene, urethane foam or the like has a property of buffering an impact by deformation. However, in general, materials such as polystyrene foam and urethane foam have low thermal conductivity. On the other hand, a highly heat conductive metal mesh such as stainless steel or a metal thin film has good heat conductivity. And deformation is easy. Therefore, it is possible to have buffer performance by the low thermal conductivity buffer material, and at the same time, to have high thermal conductivity by the metal mesh or metal thin film incorporated inside.

  A series of procedures for transporting cells using the cell transport container having the above configuration will be described.

<Step S1: Preparation>
Make the necessary preparations to transport the cells. The heat storage material box enclosing the heat storage material is packed in advance with an autoclave bag or the like, and sterilized in that state to be sterilized. The method of sterilization is autoclaving, ethylene oxide gas treatment, γ-ray irradiation, etc., and a method that does not change the properties of the heat storage material box by performing sterilization is selected. For example, when the heat storage material is hydrocarbon C ₂₀ H ₄₂ , the material of the heat storage material box is made of metal or heat-resistant polycarbonate, and the hydrocarbon C ₂₀ H ₄₂ is sealed in a completely sealed state. Since the hydrocarbon C ₂₀ H _{42 has} a melting point of 344 ° C., it does not vaporize even when subjected to autoclaving (120 ° C.), and there is no effect on the temperature maintenance performance of the heat storage material box. After sterilization, in order to store heat in the heat storage material, it is put in a thermostatic chamber in a packaged state and stored. It is desirable to install this thermostat in a room for packaging in a secondary cell transport container outside the room for cell treatment.

As the kind of the heat storage material, a pure substance having a certain melting point or a substance having a large heat capacity and a small temperature change of the melting point (for example, ± 1 ° C. or less) is used. An example is hydrocarbon. For example, when C ₂₀ H ₄₂ is used, the melting point is 36.4 ° C. Hydrocarbons with different numbers of C have different melting points. Therefore, by appropriately determining the type of hydrocarbon, it is possible to change the temperature value that the cell transport container maintains constant.

After the sterilization process, the heat storage material box in which the heat storage material is sealed is placed in a thermostatic chamber in the temperature zone to be used, and is allowed to stand until the temperature is stabilized. For example, in the case of hydrocarbon C ₂₀ H ₄₂ having a melting point of 36.4 ° C., the temperature of the thermostatic bath is 37 ° C. when most of the outside temperature to be transported is 36.4 ° C. or less. This is because, during transport, the outside temperature is lower than the melting point of C ₂₀ H ₄₂ , so heat goes out of the cell transport container. Conversely, if most of the outside temperature to be transported is 36.4 ° C or higher, the temperature of the thermostatic bath shall be 36 ° C. This is because, during transport, the outside temperature is higher than the melting point of C ₂₀ H ₄₂ , so heat enters the cell transport container from the outside.

  As with the heat storage material box, the airtight container, the culture container housing part, the heat conductive insulating material, and the like are sterilized in a packaged state. Equipment that is not resistant to various types of sterilization, such as a vacuum heat insulating material, a heat panel, first and second temperature / pressure sensors, a temperature controller, etc., shall be subjected to ethanol disinfection.

<Step S2: Carrying into cell processing facility>
Sterilized airtight container, culture container container, heat storage material box, heat conductive heat insulating material, first and second transport packaging container, culture area for cell culture in cell treatment facility (grade B clean To the degree). When moving between rooms in a cell processing facility, it is necessary to pass through a pass box in order to maintain cleanliness of the room and prevent cross contamination. When passing through the pass box, each component is sterilized by spraying ethanol from the outside of the package, placed in the pass box, and removed from the door on the side of the moving room.

  After arriving at the cell culture area, the package is opened there and removed aseptically so as not to touch the outside of the package. Since the temperature of the heat storage material box changes when exposed to room temperature, it is desirable to prepare a thermostatic chamber in the same room if possible and place it in the thermostatic chamber until use to prevent temperature changes.

  It is assumed that the first temperature / pressure sensor is preliminarily sterilized with ethanol. It is desirable to carry out sterilization and sterilization when bringing equipment into the room where cells are treated. However, since sterilization cannot be performed on mechanical devices, only ethanol treatment is generally performed.

<Step S3: Accommodation of culture vessel>
The culture vessel that has been cultured in the thermostatic chamber is moved into the safety cabinet. In the safety cabinet, equipment such as a heat block set to the same value as the temperature of the thermostatic bath is prepared in advance if necessary. Using this, the temperature of the culture container is kept constant until the culture container is completely accommodated in the cell transport container.

  Remove the lid from the culture container that has moved into the safety cabinet, and fill the culture medium into the culture container to the extent that it does not overflow. Close the lid of the culture vessel and wrap a stretchable seal such as parafilm around the boundary between the culture vessel and the lid, taking care not to spill the medium. This series of operations is performed quickly to reduce the possibility of contamination inside.

<Step S4: Accommodation in an airtight container>
The culture container packaged in step S3 is stored in the culture container storage unit prepared in advance. Next, the heat storage material box, the culture container housing part containing the culture container, and the temperature / pressure sensor are housed inside the airtight container. The temperature / pressure sensor is turned on before monitoring and starts monitoring, and measures the temperature and pressure inside the hermetic container over the entire process during transport.

<Step S5: Transporting the airtight container out of the cell processing facility>
The airtight container is taken out of the cell processing facility from the room where cell culture is performed (grade B cleanliness). When moving between rooms, in order to prevent cross-contamination, disinfect with ethanol spray and then pass through the pass box. At that time, it is desirable to cover the periphery of the airtight container with a separately prepared heat insulating material. This is because the airtight container itself does not have heat insulation performance, and heat transfer may occur between the surroundings, which may reduce the temperature maintenance performance for the culture container contained inside. is there. The wall surface inside the pass box is generally made of stainless steel. When the stainless steel wall and the airtight container come into contact with each other when passing through the pass box, the heat transfer between the inside of the airtight container is rapidly performed because the stainless steel has high thermal conductivity. For example, when the heat storage material is C ₂₀ H ₄₂ , the temperature of the pass box is room temperature (for example, 22 ° C.), so the amount of heat stored in the heat storage material flows out to the outside. Moreover, although ethanol spraying is performed for disinfection, when the ethanol sprayed vaporizes from the surface of an airtight container, air heating is taken away. This corresponds to the heat that the airtight container has lost. Considering the above, it is desirable to cover the periphery of the airtight container with a heat insulating material when moving between rooms in the cell treatment facility. Examples of the heat insulating material include polyurethane which is lightweight and sterilizable and has low thermal conductivity.

  In addition to covering with heat insulating material, there is also a means to carry the airtight container on a heater that keeps the temperature at 37 ° C. In this case, the power source of the heater needs to be portable using a dry battery or the like. Moreover, in order to perform ethanol spraying, it is necessary to have a structure in which liquid ethanol does not enter the heater.

  In step S4 and step S5, an example is shown in which the culture container packaged in step S3 is sequentially stored in the culture container storage unit and the airtight container and then carried to the room where the packaging operation is performed via the pass box. ing. Instead of this method, the packaging culture container is kept in the state of the packaged culture container using a heat insulating material or a heater, and is transported to the room where the packaging operation is carried out via a pass box, where it is sequentially stored in the culture container container and the airtight container. Also good.

<Step S6: Setting the airtight container to the secondary cell transport container>
Carry it to the room where the packaging work will be carried out and store it in the secondary cell transport container. At that time, it is desirable to temporarily store the airtight container in the thermostat installed in this room, and to store again the amount of heat lost while moving from the room for cell treatment to the room for packaging work.
A heat storage material box, an airtight container, and a second temperature / pressure sensor are installed in the secondary cell transport container, and the lid made of a vacuum heat insulating material or the like is closed. The second temperature / pressure sensor has a display that constantly displays temperature and pressure on the main body. The temperature sensor code of the second temperature / pressure sensor is installed beside the hermetic container and measures the temperature of the hermetic container. The pressure is measured outside the airtight container. A transparent window is provided in the center of the outermost lid of the cell transport container, and a second temperature / pressure sensor is installed under the transparent window. The transparent window can be opened and closed. If necessary, the transparent window is opened and the second temperature / pressure sensor is operated. Thereby, the display of the measured value of the temperature and the pressure outside the airtight container can be read without opening and closing the lid of the cell transport container during transportation. When the assembly of the cell transport container is completed, the second temperature / pressure sensor is operated, and measurement of temperature and pressure is started.

<Step S7: Transport of Cell Transport Container>
Depending on the location of the destination medical institution, the transport means is selected to transport the cell transport container. The transportation means are mainly vehicles, railroads, airplanes, and hand-carried. It is desirable to fix the cell transport container to the floor as necessary so that the cell transport container does not roll over while being transported by vehicle, railroad, or aircraft. When carrying by hand, the transport operator should pay attention so that the cell transport container does not shake as much as possible.

  Although the culture medium in the culture container is in a state of being filled in the culture container, since it cannot be completely filled, there is a slight gas phase. The effects on the cells that occur when the culture vessel is tilted during transport are that the cells enter the gas phase and dry, that the cells are affected by the surface tension created between the gas phase and the liquid phase, the liquid phase It is assumed that convection occurs in the cells and shear stress occurs in the cells. These are less affected as the gas phase portion in the culture vessel is smaller. In the transport method according to the present invention, the culture medium is filled in the culture vessel as much as possible, so that the influence of the gas phase during transport is not so great. In addition, the direction of gravity applied to the biological sample changes due to the inclination during transportation compared to the culture in the cell processing facility. As for this influence, as long as the cell transport container is upright, the direction of gravity is the same as that during culture in the cell treatment facility. Moreover, the state where the cell transport container is not upright is not taken for a long time. Therefore, the effect of gravity is considered minor.

<Step S8: Acceptance inspection at the destination>
After arriving at a medical institution or the like that is the transport destination, first, the ambient temperature and pressure of the biological sample being transported are checked. Move the monitoring device data to a PC, etc. for evaluation. Subsequently, the state of the transported biological sample is confirmed. There are various examination methods, but the sample used for treatment needs to be a non-invasive examination. That is, it is a method that does not contact a biological sample directly or via a medium or the like. Moreover, when inspecting, the culture container is once taken out from the cell transport container, evaluated quickly with a microscope, and promptly stored in the cell transport container. Thereby, the culture container is stored at the same temperature even after the inspection. In addition, about the sample which is not used for a treatment, you may investigate in detail by an invasive test | inspection. In that case, it is possible to perform various treatments on biological samples and examine the number of cells, cell viability, tissue structure, expression status of specific proteins, etc. As a result of the evaluation, it is confirmed that the transported samples are suitable for treatment If you can, start preparing for treatment. It can take up to a day to prepare for treatment. In addition, not all medical institutions have facilities such as a thermostatic bath. In that case, after arriving at the medical institution, the culture container is kept in the cell transport container and the temperature is maintained until the treatment is started.

<Step S9: Treatment>
When preparation for treatment is completed, the cell transport container is moved to a treatment room (hereinafter referred to as an operating room). Immediately before moving, stop the temperature / pressure sensor and collect the temperature and pressure monitoring data measured up to that point to a PC. Then, it is confirmed that the temperature and pressure from the departure from the cell processing facility to the point at which the storage is completed after arrival are within the allowable range. After confirmation, transport the cell transport container to the operating room. At this time, the airtight container may be taken out from the cell transport container and carried alone. A heat storage material is contained in the hermetic container, and even the hermetic container alone can maintain the internal temperature as long as it is transported from the storage room to the operating room.

When arriving at the operating room, the culture container packed in the transport packaging container is taken out. If necessary, in this state, the culture vessel is placed in a thermostatic chamber installed in the operating room and maintained at a predetermined temperature. For example, if the culture vessel has a temperature-responsive culture surface, a living body that has been subjected to a low-temperature treatment (eg, exposed to 20 ° C. for 30 minutes) and adhered to the temperature-responsive culture surface immediately prior to treatment. The sample is peeled off.
Subsequently, the biological sample is taken out from the culture container. Since the outside of the transport packaging container passes through the daily space, there is a high possibility that organisms and particles such as bacteria are attached. Therefore, it opens so that the inside of a culture container may maintain cleanliness.
First, an unclean worker in the operating room wipes the outside of the transport packaging container with a disinfectant such as ethanol or isodine. Then, the lid of the culture container is removed. Then, a clean field operator aseptically removes the biological sample using tweezers or the like so as not to contact the outside of the culture container. This is used for treatment.

101, 201 ... first heat insulating container 102, 202 ... low thermal conductivity buffer material 103, 203 ... second heat insulating container 104, 204 ... airtight container 105, 205 ... first heat storage material box 106, 206 ... culture vessel 107, 207 ... culture vessel storage part 108 ... second heat storage material box 109, 209 ... first temperature / pressure sensor 110 ... second temperature / pressure sensor 111 ... Heat insulation lid 112 ... Cord 113 ... Temperature sensor unit 114 ... Lid 115 ... Window 301 ... Air bag 401 ... Division heat storage material box 501 ... 6 well plate 502 ..1.5 ml sample tube 503... 50 ml sample tube 601... Heat storage material box 602... Heat storage material-containing bag 603. 702 ... high thermal conductivity metal mesh.

Claims (11)

A first heat insulating container having a first heat insulating material on the wall ;
A low thermal conductivity buffer material that is disposed on the inner side of the first insulating container,
A second heat insulating container disposed inside the low thermal conductivity buffer material and having a second heat insulating material on a wall surface ;
An airtight container disposed inside of the second insulating container,
And a culture container holder which is disposed inside the airtight container,
The second heat insulating material has a structure in which a heat conductive member and a buffer member having lower heat conductivity and higher buffering properties than those of the heat conductive member are alternately laminated so that the direction is random and there is no gap. A cell transport container , wherein a plurality of the cell transport containers are arranged in the container. The cell transport container according to claim 1, wherein
A cell transport container comprising an air bag filled with air in a bag together with the low thermal conductivity buffer material between the first heat insulating container and the second heat insulating container. The cell transport container according to claim 1, wherein
A cell transport container comprising a first heat storage material box enclosed with a heat storage material and disposed inside the second heat insulation container and in close contact with the airtight container. The cell transport container according to claim 1, wherein
A cell transport container comprising a second heat storage material box enclosed with a heat storage material, which is arranged inside the airtight container and in close contact with the culture container storage portion. The cell transport container according to claim 3 or 4,
A cell transport container, wherein hydrocarbon is used as the heat storage material. The cell transport container according to claim 3 or 4,
A cell transport container, wherein the first heat storage material box or the second heat storage material box is divided into a plurality of smaller divided heat storage material boxes. The cell transport container according to claim 6,
Cellular transport container, characterized by being configured at least part of the surfaces constituting the plurality of divided heat storage material box with high thermal conductivity material. The cell transport container according to claim 3, 4 or 6,
A cell transport container comprising a plurality of heat storage material-containing bags disposed in the first heat storage material box, the second heat storage material box, or a divided heat storage material box, and a heat conductive member disposed between the heat storage material-containing bags. The cell transport container according to claim 4,
The culture container storage unit and the second heat storage material box in which the heat storage material is sealed can be sterilized by sterilization, and can pass through the inside of the pass box when moving between rooms in the cell treatment facility. A cell transport container having a large size. The cell transport container according to claim 1, wherein
A cell transport container, wherein a first temperature / pressure sensor for recording changes in temperature and pressure in the hermetic container is installed inside the hermetic container. The cell transport container according to claim 1, wherein
A cell transport container, wherein a second temperature / pressure sensor for measuring and displaying the temperature and pressure outside the airtight container is installed outside the airtight container.

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