Patent Application
20100021994
The present invention relates to an apparatus for determining the termination of fat digestion and a fat tissue digestion apparatus for yielding fat-derived stem cells through digestion of a fat tissue.
Conventionally, there has been known a technique for separating fat-derived stem cells contained in fat tissue by collecting a human fat tissue and stirring it with a digestive enzyme and a physiological saline (for example, see Patent Document 1).
In this technique, the digestion goes on with stirring, and the fat tissue is decomposed to thereby separate the fat-derived stem cells. The termination of this digestion is determined by counting the time required for digestion.
Patent Document 1:
PCT International Publication No. WO 2005/012480 Pamphlet
However, the fat tissue digestion largely varies depending on the individual difference, which leads to a problem in that a group of fat-derived stem cells of constant quality can not be obtained if the termination of fat tissue digestion is determined according to the elapsed time. In addition, in order to obtain a group of sufficient fat-derived stem cells regardless of the individual difference in fat tissue, the digestion treatment needs to be performed over a long period of time to meet the time for treating a fat tissue which is slow to be digested. Thus, the treating time can not be inconveniently shortened.
The present invention takes the above situation into consideration with an object of providing an apparatus for determining the termination of fat digestion and a fat tissue digestion apparatus which are capable of yielding a group of fat-derived stem cells of constant quality regardless of the individual difference in fat tissue.
In order to achieve the above object, the present invention provides the following solutions.
A first aspect of the present invention is an apparatus for determining the termination of fat digestion for use in a fat tissue digestion apparatus which digests a fat tissue by placing and stirring the fat tissue with an enzyme solution, and any one of a physiological saline, a lactate Ringer's solution, and a buffer solution in a container, wherein the apparatus for determining the termination of fat digestion comprises: a boundary-detecting unit which detects the boundary between a fat tissue layer formed in the container by being left still after stirring and a cell suspension located below the fat tissue layer, from outside the container; a layer thickness-measuring unit which measures the thickness of the fat tissue layer based on the position of the boundary detected by the boundary-detecting unit; a fat volume-calculating unit which calculates the volume of the fat tissue layer by multiplying the thus measured thickness of the fat tissue layer by the cross-sectional area of the container; and a termination-determining unit which determines the termination of digestion based on the thus calculated volume of the fat tissue layer.
According to the first aspect, the boundary between the fat tissue and the cell suspension in the container is detected through the operation of the boundary-detecting unit, the thickness of the fat tissue layer is measured based on the position of the boundary through the operation of the layer thickness-measuring unit, and the volume of the fat tissue layer is calculated based on the thickness of the fat tissue layer and the cross-sectional area of the container through the operation of the fat volume-calculating unit. The fat tissue ends up being a cell suspension as the digestion goes on, because the tissue-connecting collagen is decomposed and thereby microstructures such as cells and extracellular tissue are lysed into a physiological saline. For this reason, the thickness of the fat tissue layer becomes thinner as the digestion goes on. Accordingly, the degree of fat tissue digestion can be accurately determined by calculating the volume of the fat tissue layer, and the termination of digestion can be determined by the operation of the termination-determining unit so that the digestion treatment can be terminated at the time when a predetermined amount of the fat tissue has been digested regardless of the individual difference in fat tissue, to yield a group of fat-derived stem cells of constant quality in a short time.
In the first aspect, the structure may be such that the container is transparent, and the boundary-detecting unit comprises a camera which is arranged to face a side wall of the container for capturing an image of the fat tissue layer inside the container through the side wall of the container.
By so doing, the boundary between the fat tissue layer and the cell suspension can be readily detected by processing the image of the fat tissue layer fat tissue layer inside the container that has been captured by the camera, and the termination of fat tissue digestion can be readily determined.
In addition, in the first aspect, the structure may also be such that the container is transparent and the boundary-detecting unit comprises a light emitting element and a light receiving element which are opposingly arranged to interpose the container.
By so doing, the boundary between the fat tissue layer and the cell suspension can be readily detected based on the amount of light received by the light receiving element out of the amount of light emitted from the light emitting element, utilizing the phenomenon that the optical absorbance differs between the fat tissue layer and the cell suspension because of their difference in the physical property.
In addition, the above structure may also be such that a plurality of the light emitting elements and the light receiving elements are arranged in a vertical direction of the container.
By so doing, even though the position of the boundary between the fat tissue layer and the cell suspension varies at the time when the termination of fat tissue digestion is determined, due to the difference in the amount of fat tissue, any one of the physiological saline, the lactate Ringer's solution, and the buffer solution placed in the container, the boundary position can be readily detected by any of the plurality of the vertically arranged light emitting elements and light receiving elements.
In addition, the above structure may also comprise a movement mechanism which moves the light emitting element and the light receiving element in the vertical direction of the container.
By so doing, even though the position of the boundary between the fat tissue layer and the cell suspension varies at the time when the termination of fat tissue digestion is determined, due to the difference in the amount of fat tissue, any one of the physiological saline, the lactate Ringer's solution, and the buffer solution placed in the container, the boundary position can be readily detected by vertically moving the light emitting element and the light receiving element through the operation of the movement mechanism.
A second aspect of the present invention is a fat tissue digestion apparatus comprising the fat digestion termination-determination apparatus of any structure mentioned above.
According to the second aspect, the amount of undigested fat tissue remaining in the container can be accurately detected through the operation of the fat digestion termination-determination apparatus, and a group of fat-derived stem cells of constant quality can be obtained.
The present invention demonstrates an effect in which a group of fat-derived stem cells of constant quality can be yielded regardless of the individual difference in fat tissue.
Hereunder is a description of a fat digestion termination-determination apparatus 1 and a fat tissue digestion apparatus 2 according to a first embodiment of the present invention, with reference to
As shown in
A fat tissue collected from a human or another mammal, a protease such as trypsin, and a physiological saline (alternatively a lactate Ringer's solution or a buffer solution) can be placed in the transparent container 3. The placed fat tissue starts float upward in the physiological saline due to the relation of their specific gravities, and forms a fat tissue layer by being left still. The volume of the fat tissue is measured in advance before the fat tissue is placed in the container, and this data is stored in a memory unit that will be described later.
As shown in
The swing mechanism 4 comprises a base 5 which swingably supports the transparent container 3, a motor 6, and a transmission mechanism 7 which is rotatably driven by the motor 6 to swing the transparent container 3 supported by the base 5. The transmission mechanism 7 comprises, for example, a pulley 7a and a belt 7b.
As shown in
The image processing unit 9 detects the position of the boundary D between the fat tissue layer A/B and the cell suspension layer C and the position of the top surface E of the fat tissue layer A/B, for example, by binarizing the captured image based on the brightness value of each pixel. Based on the position of the top surface E of the fat tissue layer A/B and the position of the boundary D detected by the image processing unit 9, the layer thickness-measuring unit 10 calculates the thickness dimension of the fat tissue layer A/B located between these positions D and E.
Then, the fat volume-calculating unit 12 calculates the volume of the fat tissue layer A/B by multiplying the cross-sectional area of the transparent container 3 that has been previously stored in the memory unit 11 by the thickness dimension of the fat tissue layer calculated by the layer thickness-measuring unit.
The digestion termination-determining unit 13 determines the termination of digestion when the thus calculated volume of the fat tissue layer A/B decreases to a predetermined ratio with respect to the stored volume of the fat tissue at the time of digestion initiation.
In order to digest the fat tissue with use of the thus configured fat tissue digestion apparatus 2 according to this embodiment, a fat tissue collected from a human or another mammal, a protease such as trypsin, and a physiological saline are placed in the transparent container 3, and the swing mechanism 4 is operated.
Through the operation of the swing mechanism 4, the transparent container 3 is swung relative to the base 5, by which the fat tissue is stirred in the protease-containing physiological saline and is digested by the enzyme. As a result, with the passage of time, collagen which connects the fat tissue contained in the free lipid layer A is decomposed and thereby microstructures such as cells and extracellular tissue are lysed into the physiological saline, by which the layer A alters into the tissue layer B, and then into the cell suspension layer C as the digestion makes further progress.
The free lipid layer A and the tissue layer B get thinner as the stirring operation makes progress through the operation of the swing mechanism 4. In this process, the determination of digestion is performed through the operation of the fat digestion termination-determination apparatus 1. Specifically, the swinging operation of the transparent container 3 with the swing mechanism 4 is halted to leave the container still for a predetermined time, for example, 5 minutes. By so doing, the fat tissue layer A/B and the cell suspension layer C are separated.
Then, an image is captured by the camera 8 arranged to face the outer surface of the side wall of the transparent container 3, and the image processing unit 9 processes the captured image to thereby detect the boundary D between the fat tissue layer A/B and the cell suspension layer C. Thereafter, the thickness of the fat tissue layer A/B is measured based on the position of the boundary D through the operation of the layer thickness-measuring unit 10, and the volume of the fat tissue layer A/B is calculated through the operation of the fat volume-calculating unit 12. Then, when the volume of the fat tissue layer A/B decreases to a predetermined ratio with respect to the volume of the fat tissue layer at the time of digestion initiation, the termination of digestion is determined through the operation of the digestion termination-determining unit 13.
In this manner, according to the fat tissue digestion apparatus 2 of this embodiment, the decreased volume of the fat tissue can be accurately measured through the operation of the fat digestion termination-determination apparatus 1. That is to say, an advantage is given in which a group of fat-derived stem cells of constant quality can be obtained regardless of the individual difference in fat tissue, as compared to conventional methods in which the termination of fat tissue digestion is determined by counting the time.
In addition, since the termination of fat tissue digestion is determined based on the image captured by the camera 8 arranged to face the outer surface of the side wall of the transparent container 3, there is no need of taking out the fat tissue from the transparent container 3 and of measuring the weight and/or the volume thereof. Thus, the volume of the fat tissue can be readily calculated in a non-contact manner. As a result, another advantage is also given in which the termination of fat tissue digestion can be automatically determined.
In this embodiment, the termination of fat tissue digestion is determined by processing the image captured by the camera 8 arranged to face the outer surface of the side wall of the transparent container 3; however, instead of this, infrared photoelectronic sensors 20 as shown in
With such a configuration, the position of the boundary D between the fat tissue layer A/B and the cell suspension C can be readily detected by utilizing the difference in the infrared absorbances of the fat tissue and the cell suspension.
In addition, in this case, as shown in
Moreover, instead of providing the plurality of the infrared photoelectronic sensors 20 in the vertical direction, the arrangement may also be such that, as shown in
Furthermore, instead of the infrared photoelectronic sensor 20, there may also be arranged, as shown in
The arrangement may also be such that, as shown in
In addition, in this embodiment, the boundary D between the fat tissue layer A/B and the cell suspension layer C is obtained, and the change in the volume of the fat tissue layer A/B is monitored based on the boundary D to determine the termination of digestion. However, instead of this, the termination of digestion may also be determined at the time when the thickness dimension of the tissue layer B decreases to a predetermined dimension or lower value by utilizing the phenomenon that the tissue layer B is reduced with the passage of time.
In the abovementioned example, the volume of the fat tissue is measured and this data is stored in the memory unit in advance before the fat tissue is placed in the container. However, rather than measuring in advance, the fat volume may also be obtained by calculation based on the measured dimension of the fat tissue layer that floats upward in the physiological saline due to the relation of their specific gravities with use of the abovementioned configuration and apparatus before the addition of the enzyme. Furthermore, the amount of the enzyme necessary for digestion of this fat volume may be calculated based on this measured result by using the calculating function of the fat volume-calculating unit, and the necessary amount of the enzyme may be added based on this calculation result.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2007-036574 | Feb 2007 | JP | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/JP2008/051721 | 2/4/2008 | WO | 00 | 8/13/2009 |
