JP6671582B2 - Method for adjusting the temperature of the contents, and a temperature control container for the contents - Google Patents

Description

  The present invention relates to a method for adjusting the temperature of contents, and a temperature control container for contents.

  In modern society, various products are distributed in large quantities, and depending on the characteristics of the products, maintaining the temperature within a certain range may be essential or preferable in order to maintain the quality. The appropriate temperature range for maintaining quality varies depending on the type and properties of the target product. For example, low-temperature distribution is often required for foods and pharmaceuticals. The temperature zone for low-temperature circulation is broadly classified into two types: freezing and refrigeration. Depending on the type and properties of the target product, refrigeration may be appropriate, but freezing may impair or degrade the quality.

  Transportation by refrigeration can be performed stably using a refrigerated transport vehicle, but transportation by refrigeration may cause a freeze accident in which products placed near the outlet of cold air may freeze. There was a problem. Further, since the refrigeration device of the refrigerated transport vehicle is normally controlled by a thermostat or the like, the temperature is repeatedly increased and decreased within a range of several degrees above and below the target temperature in a relatively short time. Depending on the type and properties of the target product to be transported, such repeated repetition of the temperature in a short time may lead to quality deterioration.

  In addition, when land, sea, and air routes are combined as in intercontinental transportation, there are transportation means that do not have low-temperature holding means, or low-temperature holding means is provided for each. However, the low-temperature distribution means at the relay point may be underdeveloped. In this case, if the transportation means without the low-temperature holding means or the relay time is long, an increase in the product temperature is unavoidable, causing a problem for an object having severe temperature conditions. Especially for high-priced products, the damage was not negligible.

  As a method of keeping the target product cold at a low temperature without using a refrigerator or freezer when transporting the target product, a method of storing a cold storage agent (cooling agent) together in a container for storing the target product is performed. ing. For example, Patent Literature 1 discloses a method of wrapping contents with a packaging material provided with a heat insulating layer and a cooling agent from the outside, and keeping the object to be transported cool. Patent Literature 2 further includes a regenerator formed by laminating two or more regenerators having different melting points from each other, and a container for accommodating the regenerator, and the outermost one of the regenerators includes a regenerator. A composite cold storage body is described in which the melting point of a cold storage auxiliary material, which is a cold storage material in the remaining layers excluding the cold storage main material, is lower than the melting point of a cold storage material of a certain cold storage main material. However, any of the methods of Patent Document 1 and Patent Document 2 is not sufficient in securing a sufficient cooling time (for example, 24 hours), and it is more difficult to cool the target product or adjust the temperature of the target product. There was a need for a practical method.

JP 2005-271949 A JP 2001-330351 A

  An object of the present invention is to provide a more practical “method for adjusting the temperature of contents” and a more practical “temperature regulating container for contents”. Specifically, the "content temperature adjustment method" and "content temperature control container" that can adjust the temperature of the content within a predetermined temperature range for a longer time, more simply and more compactly. Is to provide.

  The present inventors have conducted intensive studies, and as a result, the above-described problems have been solved by packaging the contents with a packaging material provided with a heat insulating layer, a regenerator, a latent heat transfer rate adjusting layer, and a temperature controlling agent from the outside. Can be solved, and the present invention has been completed. In particular, by providing the latent heat transfer rate adjusting layer between the regenerator and the temperature control agent, the latent heat exchange rate between the regenerator and the temperature control agent is appropriately reduced, and the content is kept within a predetermined temperature range. This makes it possible to control the temperature for a longer time and more compactly.

  In addition, the present inventors have determined that the temperature range suitable for maintaining the viability of lymphocytes as high as possible when transporting lymphocytes is 2 to 10 ° C. In order to adjust the time (for example, 24 hours), it has been found that it is preferable to use ice at −20 ° C. as a regenerator and polyethylene glycol at 10 ° C. (especially PEG-400) as a temperature regulator, and completed the present invention. I came to.

  ADVANTAGE OF THE INVENTION According to this invention, the temperature of a content can be adjusted more easily in a predetermined temperature range for a long time using a regenerator and a temperature control agent. Further, the “method for adjusting the temperature of the contents” and “the temperature-controlling container for the contents” of the present invention can have almost the same configuration regardless of the temperature change of the outside air. It can be used universally in regions with a mild climate.

Sectional drawing when the content is put in a spherical temperature control container. Sectional drawing at the time of putting a content in a rectangular parallelepiped temperature control container. It is a figure which shows the result of a "lymphocyte survival test" in Example 1 mentioned later. The vertical axis represents the number of surviving lymphocytes, and the horizontal axis represents the elapsed time since the lymphocytes were transferred to the thermostat. Of the three bar graphs at each elapsed time, the left bar graph shows the result of the thermostat at 4 ° C, the center bar graph shows the result of the thermostat at 25 ° C, and the right bar graph shows the result of the thermostat at 35 ° C. Represents It is a figure which shows the result of a "lymphocyte survival test" in Example 1 mentioned later. The vertical axis represents the number of surviving lymphocytes, and the horizontal axis represents the elapsed time since the lymphocytes were transferred to the thermostat. Of the four bar graphs at each elapsed time, the leftmost bar graph shows the results of the thermostat at 4 ° C., the second bar graph from the left shows the results of the thermostat at 10 ° C., and the third bar graph from the left. Represents the result of the thermostat at 15 ° C., and the rightmost bar graph represents the result of the thermostat at 20 ° C. It is a figure which shows the result of a "lymphocyte survival test" in Example 1 mentioned later. The vertical axis represents the number of surviving lymphocytes, and the horizontal axis represents the elapsed time since the lymphocytes were transferred to the thermostat. Of the five bar graphs at each elapsed time, the leftmost bar graph shows the result of the thermostat at 2 ° C, the second bar graph from the left shows the result of the thermostat at 4 ° C, and the third bar graph from the left. Represents the result of the thermostat at 6 ° C., the second bar graph from the right represents the result of the thermostat at 8 ° C., and the rightmost bar graph represents the result of the thermostat at 10 ° C. It is a figure which shows the result of "the temperature maintenance test of lymphocytes" (2 degreeC of thermostat) in Example 2 mentioned later. The gray area represents the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the temperature change over time of the contents. It is a figure which shows the result of "the temperature maintenance test of a lymphocyte" (5 degreeC of thermostat) in Example 2 mentioned later. The gray area represents the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the temperature change over time of the contents. It is a figure which shows the result of "the temperature retention test of a lymphocyte" (10 degreeC of thermostat) in Example 2 mentioned later. The gray area represents the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the temperature change over time of the contents. It is a figure which shows the result of the "temperature retention test of a lymphocyte" (15 degreeC of thermostat) in Example 2 mentioned later. The gray area represents the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the temperature change over time of the contents. It is a figure which shows the result of "the temperature maintenance test of a lymphocyte" (25 degreeC of thermostat) in Example 2 mentioned later. The gray area represents the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the temperature change over time of the contents. It is a figure which shows the result of "the temperature maintenance test of a lymphocyte" (35 degreeC of a thermostat) in Example 2 mentioned later. The gray area represents the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the temperature change over time of the contents. It is a figure which shows the result of the test (3 degreeC of thermostat) which becomes control of the "temperature retention test of a lymphocyte" in Example 2 mentioned later. In this control test, the latent heat transfer rate adjusting layer was not used. The gray area represents the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the temperature change over time of the contents. It is a figure which shows the result of the test (35 degreeC of thermostat) which becomes control of the "temperature retention test of a lymphocyte" in Example 2 mentioned later. In this control test, the latent heat transfer rate adjusting layer was not used. The gray area indicates the target temperature zone (2 to 10 ° C.). The temperature rises to about 35 ° C. immediately after the start of the test, and thereafter, the line that maintains approximately 35 ° C. indicates the temperature over time in the thermostat. The smooth curve represents the temperature change of the contents over time.

<How to adjust the temperature of the contents>
The “method of adjusting the temperature of contents” of the present invention (hereinafter, also simply referred to as “the adjusting method of the present invention”) includes a heat insulating layer, a regenerator, a latent heat transfer speed adjusting layer, and a temperature regulator from the outside. There is no particular limitation as long as the method includes wrapping the contents in a packaging material and controlling the temperature. Such contents are not particularly limited as long as the contents are to be adjusted, and examples thereof include biological samples for medical use, medicines, and foods. Examples of the above-mentioned biological sample for medical use include cells such as lymphocytes and iPS cells; blood; organs; and the like, among which cells such as lymphocytes and iPS cells are preferable, and lymphocytes are more preferable. Can be

  The above-mentioned "heat insulating layer" is a layer containing a heat insulating material, and is preferably a layer made of a heat insulating material. The material of the heat insulating material is not particularly limited, and is a resin foam heat insulating material such as expanded polystyrene, polyurethane foam, or polyethylene; a fiber heat insulating material such as glass wool or rock wool; cork, cellulose, cellulose fiber, wool, or the like. Natural material-based heat insulating materials; vacuum heat insulating panels; As these heat insulating materials, commercially available materials can be used.

  The material of the heat insulating layer may be one type, or two or more types may be used in combination. Preferred examples of the heat-insulating layer include a paper powder (weight ratio of 51% or more), a foam made of starch and polypropylene (M's cushion: Mutohunipack Co., Ltd.). In addition, the heat insulating layer may be composed of a single layer, or may be composed of two or more layers. The thickness of the heat insulating layer is not particularly limited, but may be in the range of 1 to 10 cm.

  In the present invention, “cooling agent” means a substance having a melting point of 0 ° C. or less. As such a regenerator, ice (melting point: 0 ° C.) is preferred because of its large latent heat and low cost. Since ice has a large latent heat, the temperature of the content can be adjusted more compactly, for example, as compared to a case where the same content is temperature-controlled for the same time by using only a thermostat. Further, the layer of the regenerator may be composed of a single layer, or may be composed of two or more layers of the regenerator. Preferable cold storage agents include, for example, Chillmates (Ice Japan Co., Ltd.).

  The regenerator at the start of the adjusting method of the present invention may be a regenerator in a solid-liquid co-phase, but from the viewpoint of adjusting the temperature of the contents within a predetermined temperature range for a longer time, , And more preferably a solid-phase regenerator adjusted to a temperature 5 to 25 ° C. lower than the melting point of the regenerator.

  In the adjusting method of the present invention, as the regenerator, those stored in a container such as a bag made of resin or the like can be suitably used. For example, the aforementioned chill mates (Ice Japan Co., Ltd.) are housed in a rayon-based nonwoven bag. As the regenerator stored in the container, a commercially available regenerator can be used.

  The “latent heat transfer speed adjusting layer” is a layer containing a heat insulating material, and is preferably a layer made of a heat insulating material. The “latent heat transfer speed adjusting layer” is disposed between the regenerator and the temperature regulator. Examples of the material of the heat insulating material include the same materials as those of the heat insulating material of the heat insulating layer. Further, in order to prevent the surface of the container of the cold storage agent from becoming wet due to dew condensation and the like, thereby preventing the contamination of the product and the deterioration of the heat insulation performance, the container of the cold storage agent is wrapped with a resin buffer material (preferably a bubble buffer material). Can be suitably used. Since such a resin buffer material (preferably a bubble buffer material) has a heat insulating effect, it can also be used as a latent heat transfer speed adjusting layer. The degree to which the cold storage agent is wrapped by the buffer material is not particularly limited, and the cold storage agent may be wrapped in a single layer or in two or more layers by the buffer material, and may be appropriately adjusted according to an appropriate latent heat transfer speed. it can. Preferable bubble cushioning materials include, for example, bubble wrap (Kawakami Sangyo Co., Ltd.). As the above-mentioned heat insulating material and cushioning material, commercially available materials can be used.

  The material of the latent heat transfer speed adjusting layer may be one type, or two or more types may be used in combination. Further, the latent heat transfer speed adjusting layer may be composed of a single layer, or may be composed of two or more layers. The material of the heat insulating material of the heat insulating layer and the material of the heat insulating material of the latent heat transfer speed adjusting layer may be the same or different.

  Further, from the viewpoint of adjusting the temperature of the contents within a predetermined temperature range for a longer time, the thickness and the material of the latent heat transfer speed adjustment layer are adjusted so that the latent heat exchange speed between the regenerator and the temperature regulator becomes appropriate. Adjustment is preferred. If the above-mentioned latent heat exchange rate is too high, the latent heat of the regenerator will rapidly shift to the temperature control agent, and may fall below the temperature lowering limit temperature of the contents. This is because the latent heat for transferring from the agent to the temperature control agent may be insufficient, and may exceed the temperature control upper limit temperature of the contents. The adjustment of the latent heat exchange rate can be performed by, for example, measuring the temperature of the content over time.

  In the present invention, “temperature controlling agent” means a substance having a melting point exceeding 0 ° C., and such a temperature controlling agent can suitably include a substance having a melting point exceeding 0 ° C. and 25 ° C. or less. . Specific examples of the temperature regulator include polyethylene glycol 400 (melting point 4 to 8 ° C), polyethylene glycol 600 (melting point 15 to 25 ° C), n-pentadecane (melting point 6 ° C), and n-tetradecane (melting point 10 ° C). N-hexadecane (melting point 17 ° C), n-heptadecane (melting point 22 ° C), n-octadecane (melting point 28 ° C), caprylic alcohol (melting point 7 ° C), lauryl alcohol (melting point 24 ° C), octanoic acid (melting point 17 ° C) ), Methyl laurate (melting point 5 ° C.), methyl myristate (melting point 19 ° C.), and the like. The temperature controlling agent may be a single kind of substance or a mixture of plural kinds of substances. In the case of a mixture of plural kinds of substances, a temperature range can be given to the melting point. Further, the layer of the temperature controlling agent may be constituted by a single layer, or may be constituted by two or more layers of the temperature controlling agent.

  In the adjusting method of the present invention, as the temperature adjusting agent, those contained in a container such as a bag made of resin or the like can be suitably used. For example, PEG-400 (Sanyo Chemical Industries, Ltd.) is housed in a plastic bag.

  In addition, in order to prevent the surface of the container of the temperature control agent from becoming wet due to dew condensation and the like, and thereby to prevent the contamination of the product and the deterioration of the heat insulation performance, the container of the temperature control agent is made of a resin buffer material (preferably a bubble buffer material). Wrapped ones can be suitably used. The resin buffer material (preferably a bubble buffer material) that wraps the temperature control agent has a heat insulating effect, and therefore can be used also as a latent heat transfer speed control layer between the regenerator and the temperature control agent. . There is no particular limitation on the degree of wrapping the temperature control agent in the buffer material, and the temperature control agent may be wrapped in one layer or in two or more layers with the buffer material, and adjusted appropriately according to an appropriate latent heat transfer speed. be able to. Preferable bubble cushioning materials include, for example, bubble wrap (Kawakami Sangyo Co., Ltd.). As the above-mentioned temperature regulator and buffer material, commercially available materials can be used.

  In the adjusting method of the present invention, it is preferable to use a temperature control agent having a melting point near the temperature of the content. By using such a temperature control agent, the content can be adjusted to a temperature close to the temperature control temperature for a longer time. The temperature control agent having a melting point near the temperature of the content can be appropriately selected based on the above description of the temperature control agent and its melting point and known information.

  In the adjustment method of the present invention, a preferable temperature control temperature is determined by the properties of the content. However, the adjustment method of the present invention adjusts the temperature of the content to any temperature range included in the range of 2 to 25 ° C, for example. It can be suitably used for adjustment. Examples of such a temperature zone include a temperature zone within plus or minus 3 ° C. with respect to the temperature of the melting point of the temperature controlling agent used.

  The temperature control agent at the time of starting the adjustment method of the present invention may be any of a solid phase, a solid-liquid co-phase, and a liquid phase, but a viewpoint of adjusting the temperature of the content within a predetermined temperature range for a longer time. From the above, it is preferable that the temperature regulator is in a solid-liquid co-phase or liquid phase, and it is more preferable that the temperature regulator is in a liquid phase. It is further preferable that the thermostat is in a liquid phase, which is controlled to a temperature higher by ° C.

  The preferable ratio of the amount of the regenerator to the amount of the regenerator used in the adjustment method of the present invention depends on the kind of the regenerator used and how much the temperature to be adjusted is set, etc. Although it is not possible, the weight ratio between the regenerator and the temperature regulator used is 2: 1 to 1: 8, 1.5: 1 to 1: 6, or 1: 1 to 1: 4. Further, the preferable ratio of the amount of the regenerator to the amount of the regenerator used in the adjusting method of the present invention is such that when the regenerator becomes a liquid phase after a lapse of time, the reconstituter becomes solid-liquid co-phase. Alternatively, it is preferable that the ratio is such that the contents are in a solid phase from the viewpoint of adjusting the temperature of the contents within a predetermined temperature range for a longer time. Such a preferable ratio can be appropriately determined.

  The packaging material used in the adjusting method of the present invention includes a heat insulating layer, a regenerator, a latent heat transfer speed adjusting layer, and a temperature regulator from the outside, but as another configuration, one or more layers of another regenerator, It may further include one or more layers selected from the group consisting of one or more layers of another temperature control agent and one or more heat insulating layers. As another configuration, for example, a latent heat transfer speed adjusting layer may be further provided inside the temperature adjusting agent. As a packaging material having another configuration, for example, a heat insulating layer 1, a heat insulating layer 2, a regenerator 1, a latent heat transfer speed adjusting layer 1, a latent heat transfer speed adjusting layer 2, a temperature control agent, and a latent heat transfer speed adjusting layer 3 are provided from the outside. Wrapping material, or a packaging material provided with a heat insulating layer 1, a regenerator 1, a heat insulating layer 2, a regenerator 2, a latent heat transfer speed adjusting layer, and a temperature regulator from the outside, or a heat insulating layer 1, a regenerator 1, and a heat insulating material from the outside. Layer 2, cold storage agent 2, latent heat transfer speed adjustment layer 1, temperature control agent 1, latent heat transfer speed adjustment layer 2, packaging material provided with temperature control agent 2, and the like.

  In the adjusting method of the present invention, the content is temperature-controlled by packaging the content with the above-mentioned packaging material. The method and mode of packaging the content with such a packaging material are not particularly limited. For example, as shown in FIG. 1, the content may be spherically packaged with the packaging material such that the content is substantially at the center. As shown in FIG. 2, the contents may be packaged in layers with a packaging material so as to sandwich the contents.

  The contents packaged with the packaging material need not be stored in the container, but are preferably stored in the container from the viewpoint of easy transportation. Such a container may be, for example, a container that is not made of a heat insulating material like a normal cardboard box, or may be a container that is made of a heat insulating material. When using a container made of a heat insulating material, the heat insulating material on the outer wall of the container may be used as the heat insulating layer of the packaging material in the present invention.

  In the adjusting method of the present invention, the content is kept within a predetermined temperature range, for example, 15 hours or more, preferably 18 hours or more, more preferably 20 hours or more, further preferably 22 hours or more, more preferably 24 hours or more, It can be maintained for preferably at least 28 hours, more preferably at least 32 hours. Couriers in Japan can deliver packages to their destinations in about 24 hours, and can travel a significant distance in about 24 hours using other means of transportation. Therefore, the technical significance of maintaining the contents within a predetermined temperature range for about 24 hours is significant.

  As a more specific embodiment of the adjusting method of the present invention, a preferred embodiment in the case where the content is adjusted to 2 to 10 ° C is described below. The temperature range of 2 to 10 ° C. is a temperature range suitable for maintaining the viability of lymphocytes as high as possible. In this embodiment, ice (melting point: 0 ° C.) (preferably chillmates (Ice Japan Co., Ltd.)) at −25 ° C. to −15 ° C. (preferably −20 ° C.) is used as a regenerator, and 7 ° C. PEG-400 having a melting point of 4 to 8 ° C (preferably 10 ° C) (preferably PEG-400 Sanyo Chemical Industries, Ltd. stored in a plastic bag) is preferably exemplified. Moreover, as a preferable weight ratio of the regenerator and the temperature regulator used in such an embodiment, a weight ratio of preferably 1: 1.5 to 1: 2.5, and 1: 1.8 to 1: 2.2 is preferable. More preferably. Further, in such an embodiment, the above-described regenerator and the above-mentioned temperature regulator are each a bubble cushioning material (preferably bubble wrap (Kawakami Sangyo Co., Ltd.)), and the regenerator is double-wrapped and the temperature regulator is single-wrapped. Are preferred. In this embodiment, the heat-insulating layer may be a foam having a thickness of 1 to 10 cm, preferably a foam made of paper powder (weight ratio of 51% or more), starch and polypropylene (M's cushion: K.K. Mutoh Unipack) (4 cm thick). In these embodiments, at the time when all the regenerator becomes liquid phase after a lapse of time, the temperature control agent has a ratio of a solid-liquid co-phase or a solid phase. It is preferable from the viewpoint of adjusting for a longer time within the range.

<Content temperature control container>
The “temperature-controlling container for contents” of the present invention (hereinafter, also simply referred to as “temperature-controlling container of the present invention”) includes a heat insulating layer, a regenerator, a latent heat transfer speed adjusting layer, and a temperature-controlling agent from the outside. The container is not particularly limited as long as it is a container.

  Although the shape of the container is not particularly limited, a substantially rectangular parallelepiped shape is preferable. The material of the container is not particularly limited, and examples thereof include metal, cardboard, plastic, and wooden. The container is not made of a heat insulating material and may not be provided with a heat insulating material, but is preferably made of a heat insulating material or provided with a heat insulating material. Further, as the above-mentioned container, containers of the same or different materials may be used in double or triple or more. For example, a container made of cardboard may be further used in a plastic container.

<Other inventions>
Aspects of the present invention also include a "temperature control material for contents." The temperature control material for the contents is not particularly limited as long as it is a packaging material provided with a heat insulating layer, a regenerator, a latent heat transfer speed control layer, and a temperature control agent from the outside. If the content is wrapped with such a packaging material, the temperature of the content can be adjusted, and if the packaging material is provided in a container, a temperature-controlled container for the content can be manufactured.

[Reference example]
[Method for preparing lymphocytes]
Lymphocytes used in Examples described later were prepared by the following method.

"Preparation of peripheral blood lymphocyte cells"
Peripheral blood (100 mL) of one healthy subject with consent was collected by intravenous addition of heparin. The collected blood sample was transferred to a 250 mL centrifuge tube (BD Falcon; 352075), 100 mL of a washing solution (Nikken Biomedical Research Institute) was added, and the mixture was gently mixed and diluted twice. The following operations including this operation were performed aseptically. Two-fold diluted blood was layered on five 50 mL centrifuge tubes (BD Falcon; 352070) into which 10 mL of Ficoll (manufactured by GE Healthcare Bioscience) was dispensed, and the centrifuge tubes were placed at 1800 rpm for 20 minutes at room temperature. Centrifugation was performed without applying a brake (Centrifuge: H-700 manufactured by Kokusan Co., Ltd.). The mononuclear cell layer of the intermediate layer was collected in two 50 mL centrifuge tubes, and an equal amount of a washing solution was added to each. The centrifuge tube was closed with the lid closed, mixed by inverting 2-3 times, and centrifuged at 1800 rpm for 15 minutes at room temperature. After centrifugation, the supernatant was removed, and 40 mL of RPMI1640 + 9Sin (manufactured by Lymphotech) (10% human serum (mannheim research institute)) and 700 U / mL rIL-2 (proleukin: Novartis) were added to the obtained pellet. ) To give a cell suspension.
40 μL of Turk's solution (manufactured by Mutoh Chemical Co., Ltd.) was dispensed into a 1.5 mL Eppendorf tube (manufactured by Eppendorf), and 10 μL of the cell suspension was added thereto and mixed, and 10 μL of the mixed solution was used in a Neubauer hemocytometer. (Elmer; 9731), and the number of cells was calculated under a microscope (Olympus Optical Co., Ltd .; 211320).
Further, 20 μL of trypan blue staining solution (manufactured by SIGMA; 93595) was dispensed into a 1.5 mL Eppendorf tube, 10 μL of the cell suspension was mixed with the solution, and 10 μL of the mixed solution was poured into a Neubauer hemocytometer. Unstained living cells were counted under a microscope, and the survival rate was calculated.

"Preparation of OKT3 solid-phase flask"
A solution of OKT3 (importer: Janssen Kyowa Co., Ltd., manufacturer: Osofa Chemical: OKT3) was prepared with physiological saline to a concentration of 5 μg / mL, and 10 mL of the prepared solution was cultured in a 225 cm ² bottom culture flask (Sumitomo Bakelite Co., Ltd.). (Manufactured by the company), and the entire bottom surface was covered with an OKT3 solution. After leaving still for 3 hours or more, the OKT3 solution in the flask was sucked with a suction device, about 100 mL of physiological saline solution (manufacturer: Otsuka Pharmaceutical Co., Ltd.) was poured, the lid of the flask was closed, and after vigorously shaking, the liquid in the flask was discarded. Again, about 100 mL of a physiological saline solution (manufacturer: Otsuka Pharmaceutical Co., Ltd.) was poured into the flask, and allowed to stand at room temperature for 15 minutes with the solid-phased surface facing down. Thereafter, the lid was closed and the mixture was shaken vigorously for 1 minute, and the liquid contained therein was discarded. The liquid remaining in the flask and on the lid was carefully sucked with a suction machine to obtain an OKT3 solid-phased flask. When not used on the day of preparation, it was stored at 4 ° C. until use, leaving a small amount of physiological saline.

(Culture of peripheral blood lymphocyte cells)
The cell suspension prepared in the above “Preparation of Peripheral Blood Lymphocyte Cells” was dispensed by 10 mL each into four 50 mL centrifuge tubes, and RPMI1640 + 9Sin (manufactured by Lymphotech) (10% human serum (Mannheim research) (Incl. 700 U / mL rIL-2 (proleukin: Novartis)). The entire amount of each was seeded on a 225 cm ² flask with an OKT3 solid phase prepared in the above (Preparation of an OKT3 solid phase flask), and the temperature was 37.0 ± 0.5 in a Teher type CO ₂ incubator (manufactured by Hirasawa). Cultivation was carried out at a temperature of 95.0 ± 5.0% and a CO ₂ concentration of 5.0 ± 0.2%. During the culture period, the above-described medium was added as appropriate, and the culture was continued for 14 days. At the end of the culture, the culture flask is tapped to collect lymphocyte cells attached to the bottom, and the total lymphocyte cell count and viability are measured in the same manner as in the above "Preparation of peripheral blood lymphocyte cells".・ Calculated.

  Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited to these examples.

[Examination of temperature range suitable for maintaining lymphocyte viability]
The following “lymphocyte survival test” was performed to examine a temperature range suitable for maintaining the lymphocyte viability as high as possible when transporting lymphocytes.

(Method of lymphocyte survival test)
1 × 10 ⁹ lymphocytes obtained by culturing according to the preparation method of Reference Example were suspended in 210 mL of physiological saline to which albumin was added, and the entire amount of the lymphocyte suspension was placed in a separation bag (manufactured by Terumo). Moved to Three such separation bags containing the lymphocyte suspension were prepared for each temperature zone. After each of the thermostats (manufactured by Sanyo Electric Co., Ltd .; MIR-153) was set to a predetermined temperature, the above-mentioned separation bag containing the lymphocyte suspension was transferred to each thermostat. At 24 hours, 36 hours, 48 hours, and 72 hours after transfer to the thermostat, the cell numbers were measured.

(Result 1 of survival test of lymphocyte)
FIG. 3 shows the results of the above-described lymphocyte survival tests, with the temperature of the thermostat set at 4 ° C., 25 ° C., and 35 ° C., respectively. As can be seen from FIG. 3, when the lymphocytes were kept at 4 ° C., about 75% of the survival rate could be maintained after 24 hours, whereas when the lymphocytes were kept at 25 ° C., the survival rate after 24 hours. Was reduced to about 40%, indicating that the survival rate after 24 hours was reduced to about 25% when the lymphocytes were kept at 35 ° C. From these results, it was shown that 4 ° C. was suitable for maintaining lymphocytes, but 25 ° C. and 35 ° C. were not suitable for maintaining lymphocytes.

(Results of lymphocyte survival test 2)
FIG. 4 shows the results of the above-described lymphocyte survival tests, with the temperature of the thermostat set at 4 ° C., 10 ° C., 15 ° C., and 20 ° C., respectively. As can be seen from FIG. 4, when the lymphocytes were kept at 4 ° C. or 10 ° C., a survival rate of about 70% could be maintained after 24 hours, and when the lymphocytes were kept at 15 ° C., about 60% of the survival rate was maintained after 24 hours. While the survival rate was maintained, it was shown that the survival rate after 24 hours was reduced to less than about 50% when the lymphocytes were kept at 20 ° C. These results indicate that 4 to 15 ° C is preferable and 4 to 10 ° C is more preferable for maintaining lymphocytes.

(Results of lymphocyte survival test 3)
FIG. 5 shows the results of the above-described lymphocyte survival tests, with the temperature of the thermostat set at 2 ° C., 4 ° C., 6 ° C., 8 ° C., and 10 ° C., respectively. As can be seen from FIG. 5, a survival rate of about 80% can be maintained after 24 hours, and a survival rate of about 65% to about 80% can be maintained even after 36 hours, regardless of the temperature maintained. It has been shown. These results indicate that 2 to 10 ° C is preferable and 4 to 10 ° C is more preferable for maintaining lymphocytes.

[Examination of transport mode for keeping lymphocytes at 2 to 10 ° C for a long time]
The experiment of Example 1 showed that it is preferable to keep the lymphocytes at 2 to 10 ° C. in order to maintain the viability of the lymphocytes. Therefore, the following “lymphocyte temperature retention test” was performed in order to examine a transport mode for keeping lymphocytes at 2 to 10 ° C. for a long time.

(Method of temperature retention test for lymphocytes)
One separation bag containing a lymphocyte suspension similar to that used in Example 1 was prepared for each test group. In order to be able to measure the temperature of the separation bag over time, the one placed in a plastic bag with the sensor part of the thermometer kept in contact with the outer surface of the separation bag was defined as the "content" used in this test. The content was adjusted to 2 ° C. As the heat insulating layer, a foam (M's cushion: Mutohunipack Co., Ltd.) made of paper powder (weight ratio of 51% or more), starch and polypropylene and having a thickness of 4 cm was used. The ice (0.5 kg or 0.75 kg) contained in the bag was adjusted to -10 ° C or -20 ° C (Chillmates: Ice Japan Co., Ltd.). The PEG-400 (0.5 kg or 1.0 kg) (Sanyo Chemical Industry Co., Ltd.) containing the PEG-400 and adjusted to 5 ° C., 10 ° C. or 15 ° C. was used. As the latent heat transfer speed adjusting layer, a bubble cushioning material (bubble wrap: Kawakami Sangyo Co., Ltd.) made of polyethylene is used. A warming agent was used for each. Further, as a container, a rectangular parallelepiped cardboard box is prepared, and a heat insulating layer, a cold storage agent, a latent heat transfer speed adjusting layer, and a temperature control agent are arranged in the above-described cardboard box so as to have a sectional view as shown in FIG. Then, a temperature control container of the present invention was prepared, and the contents were arranged between temperature control agents. After placing the temperature-regulating vessel containing the contents in a thermostat, the temperature of the contents was measured over time.

(Results of lymphocyte temperature retention test 1)
The above-mentioned temperature maintenance test of lymphocytes was performed, and the time during which the contents could be maintained within a temperature range of 2 to 10 ° C. was compared. As a result, 0.5 kg of a cooling agent adjusted to −20 ° C. was used. It turned out that it is preferable to use 1 kg of a warming agent adjusted to 10 ° C. Therefore, in the following “lymphocyte temperature retention test”, the method of this preferred embodiment was adopted.

(Result 2 of lymphocyte temperature retention test)
After changing the set temperature of the thermostat in various ways, the result of performing the “lymphocyte temperature retention test” is shown in FIGS. 6 (2 ° C thermostat), FIG. 7 (5 ° C thermostat), and FIG. 10 (15 ° C.), FIG. 10 (25 ° C.), and FIG. 11 (35 ° C.). The gray areas in FIGS. 6 to 11 represent the target temperature zone (2 to 10 ° C.), the zigzag line represents the temperature change over time in the thermostat, and the smooth curve represents the content over time. Indicates temperature change. 6 to 11 show the temperature change from the start of the temperature holding test to the elapse of 24 hours. The temperature at the right end of the graphs in FIGS. 6 to 11 indicates the temperature of the contents after 24 hours.

  As can be seen from FIGS. 6 to 11, when the temperature control container of the present invention is used, regardless of the temperature of the outside air of the temperature control container (that is, the temperature of the thermostat), the temperature range is set to the target temperature zone (2 to 10 ° C.). It was shown that the temperature of the contents could be maintained for a long time. When the temperature of the thermostat was 5 ° C., 10 ° C., 15 ° C., and 25 ° C., the temperature of the contents could be maintained in the range of 2 ° C. to 10 ° C. even after 24 hours. When the temperature of the thermostat is 2 ° C., the temperature of the contents after 24 hours is slightly lower than 2 ° C., but the amount of the regenerator is slightly reduced and / or the amount of the thermoregulator is reduced. For example, if the temperature of the contents is slightly increased, the temperature of the contents after 24 hours has elapsed can be maintained in the range of 2 to 10 ° C. When the temperature of the thermostat is 35 ° C., the temperature of the contents after 24 hours is slightly higher than 10 ° C. However, if the amount of the regenerator is slightly increased, for example, at the time after 24 hours, Can be maintained in the range of 2 to 10 ° C.

  In addition, as a control, except that the latent heat transfer speed adjustment layer was not provided, and that the thermostat was set at 3 ° C. or 35 ° C., the results of the test performed in the same manner as the “lymphocyte temperature retention test” described above. Are shown in FIG. 12 (constant temperature bath 3 ° C.) and FIG. 13 (constant temperature bath 35 ° C.). A comparison between FIG. 12 (control, constant temperature bath 3 ° C.) and FIG. 6 (constant temperature bath 2 ° C.) and FIG. 7 (constant temperature bath 5 ° C.) shows that the control without the latent heat transfer rate adjusting layer was 4 hours after the start of the test. The temperature of the contents became 0 ° C. or less at about the same time, and the target temperature zone (2 to 10 ° C.) could be maintained for only about 3 hours. In addition, comparing FIG. 13 (control, constant temperature bath 35 ° C.) with FIG. 11 (constant temperature bath 35 ° C.), in the control without using the latent heat transfer rate adjusting layer, after a certain period of time has elapsed from the start of the test, It has been shown that the temperature rises rapidly and deviates rapidly from the target temperature zone. From these results, it was found that the latent heat transfer rate adjusting layer plays an important role.

  As described above, it has been shown that the temperature control container of the present invention can maintain the temperature of the contents in the target temperature zone for a long time without being affected by the temperature of the outside air of the temperature control container.

  INDUSTRIAL APPLICABILITY The present invention can be suitably used in the field of adjusting the temperature of contents within a predetermined temperature range for a longer time, more simply, and more compactly using a regenerative agent or a temperature controlling agent.

Claims (18)

Thermal insulation layer from the outside, the refrigerant 13A, latent migration rate adjusting layer, the temperature adjustment agent, the temperature adjusting method of the contents, characterized in that packaged tempering the content at packaging material having a latent heat migration speed adjustment layer And
The adjustment method, wherein the temperature control agent at the time of starting the adjustment method is a solid-liquid co-phase or a liquid phase . The method according to claim 1, wherein the heat insulating layer is a foam made of paper powder, starch, and polypropylene. The method according to claim 1 or 2, wherein the regenerator is ice at -10 to -20C. The method for adjusting the temperature of contents according to any one of claims 1 to 3, wherein the latent heat transfer speed adjusting layer is a foam cushioning material made of polyethylene. The method for adjusting the temperature of contents according to any one of claims 1 to 4, wherein the temperature adjusting agent is PEG-400. The method for adjusting the temperature of contents according to any one of claims 1 to 5, wherein the contents are lymphocytes. The method for adjusting the temperature of contents according to any one of claims 1 to 6, wherein the temperature control is performed within a predetermined temperature range. The method according to claim 7, wherein the temperature is adjusted within a temperature range of 2 to 10C. The method for adjusting the temperature of contents according to any one of claims 1 to 8, wherein the temperature is adjusted without being affected by the outside air temperature. From the outside, a heat insulating container, a regenerator, a latent heat transfer speed adjustment layer, a temperature control agent , a temperature control container for the contents characterized by comprising a latent heat transfer speed adjustment layer ,
The content temperature control container, wherein the temperature control agent at the time of starting temperature control of the content in the content temperature control container is a solid-liquid co-phase or a liquid phase . 11. The content temperature control container according to claim 10, wherein the heat insulating layer is a foam made of paper powder, starch, and polypropylene. The temperature control container for contents according to claim 10 or 11, wherein the regenerator is ice at -10 to -20 ° C. The temperature control container for contents according to any one of claims 10 to 12, wherein the latent heat transfer speed adjusting layer is a foam cushioning material made of polyethylene. The temperature control container for contents according to any one of claims 10 to 13, wherein the temperature control agent is PEG-400. The content-controlling container according to any one of claims 10 to 14, wherein the content is a lymphocyte. The temperature control container for contents according to any one of claims 10 to 15, wherein the temperature of the contents is controlled within a predetermined temperature range. 17. The content temperature control container according to claim 16, wherein the temperature of the content is controlled within a temperature range of 2 to 10C. The temperature control container for contents according to any one of claims 10 to 17, wherein the temperature of the contents is controlled without being affected by the outside air temperature.