JP2009095565A - Body tissue filling material and its manufacturing method - Google Patents

Body tissue filling material and its manufacturing method Download PDF

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JP2009095565A
JP2009095565A JP2007271575A JP2007271575A JP2009095565A JP 2009095565 A JP2009095565 A JP 2009095565A JP 2007271575 A JP2007271575 A JP 2007271575A JP 2007271575 A JP2007271575 A JP 2007271575A JP 2009095565 A JP2009095565 A JP 2009095565A
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tissue
supernatant
biological tissue
filling material
step
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JP5094327B2 (en
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Hironori Tsuchida
裕基 土田
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Olympus Corp
オリンパス株式会社
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Abstract

An object of the present invention is to have a high fixability to a living tissue defect or the like and to be transplanted easily without using a syringe or the like.
A cell suspension obtained by digesting a living tissue is centrifuged (step S2), the separated erythrocyte component is removed (step S3), and the cell suspension after removal of the erythrocyte component is again treated. By centrifuging (step S4), removing a part of the separated supernatant (step S5), and adding calcium chloride to the liquid containing the tissue-derived cells and platelet components after removing the supernatant (step S6) Provided is a gel-like biological tissue filling material containing the tissue-derived cells produced.
[Selection] Figure 1

Description

  The present invention relates to a biological tissue filling material and a method for producing the same.

  Conventionally, human adipose tissue is collected, digested by stirring with digestive enzymes and physiological saline, and adipose-derived cells are extracted by centrifugal concentration of the resulting cell suspension. (For example, refer to Patent Document 1).

International Publication No. 2005/012480 Pamphlet

  However, the adipose tissue-derived cells obtained in this way have high fluidity, and as such, even if transplanted into a living tissue defect part or the like in a living body, the cells will flow without being settled. There is an inconvenience that the biological tissue forming action cannot be promoted. In addition, in order to transplant fat tissue-derived cells with high fluidity, it is premised that a syringe, a catheter, or the like is used.

  The present invention has been made in view of the circumstances described above, and provides a biological tissue filling material that is highly fixable to a biological tissue defect or the like and can be easily transplanted without using a syringe or the like, and a method for manufacturing the same. It is intended to provide.

In order to achieve the above object, the present invention provides the following means.
The present invention involves centrifuging a cell suspension obtained by digesting a biological tissue, removing the separated erythrocyte component, centrifuging the cell suspension after removing the erythrocyte component again, Provided is a gel-like biological tissue filling material containing tissue-derived cells produced by removing a part of the supernatant and adding calcium chloride to a liquid containing tissue-derived cells and platelet components after removal of the supernatant.

  According to the present invention, the cell suspension contains tissue-derived cells and blood components other than red blood cells by centrifuging and removing red blood cell components from the cell suspension obtained by digesting the biological tissue. It comes to be. Subsequently, the cell suspension obtained in this manner is centrifuged again, whereby a liquid containing tissue-derived cells and a platelet component is allowed to settle, and other blood components can be separated as a supernatant.

  And, by adding calcium chloride to a liquid containing tissue-derived cells and platelet components obtained by removing a part of the supernatant, the thrombin and calcium chloride contained in the supernatant react, It becomes a gel-like biological tissue filling material. The biological tissue filling material thus obtained is in a gel form with low fluidity, and therefore, when transplanted into a biological tissue defect or the like, it is fixed in place without flowing. Accordingly, it is possible to fix the tissue-derived cells contained in the biological tissue defect and grow it in situ to promote the repair of the biological tissue.

Moreover, the growth of living body-derived cells is promoted by the contained platelet component, and early repair of living tissue can be achieved.
Furthermore, since it is a gel with low fluidity, it is not limited to syringes and catheters, and can be transplanted using other instruments and methods.

In the above invention, the living tissue may be a fat tissue.
If it is an adipose tissue, it can be abundantly collected from a patient, and its manufacture is easy.

  The present invention also provides a first centrifugation step for centrifuging erythrocyte components from a cell suspension obtained by digesting living tissue, and removing the erythrocyte components separated by the first centrifugation step. An erythrocyte removal step, a second centrifugation step for concentrating tissue-derived cells and platelet components from the cell suspension after removal of the erythrocyte component by the erythrocyte removal step, and centrifugation after the second centrifugation step Production of a biological tissue filling material comprising: a supernatant removal step for removing a portion of the supernatant; and an addition step for adding calcium chloride to a liquid containing tissue-derived cells and platelet components after removal of the supernatant in the supernatant removal step Provide a method.

According to the present invention, in the process of digesting a living tissue, a gel-like living tissue filling material is obtained by leaving a portion containing a platelet component and thrombin without removing all blood components and adding calcium chloride. Can be easily manufactured.
In the above invention, the living tissue may be a fat tissue.

According to the biological tissue filling material according to the present invention, the fixability to a biological tissue defect portion or the like is high, and there is an effect that it can be easily transplanted without using a syringe or the like.
Moreover, according to the manufacturing method of the biological tissue filling material which concerns on this invention, in the extraction process of the tissue origin cell from a biological tissue, there exists an effect that a gel-like biological tissue filling material can be manufactured easily.

A biological tissue filling material and a manufacturing method thereof according to an embodiment of the present invention will be described below with reference to FIGS.
The biological tissue filling material according to the present embodiment is, for example, a gel-like biological tissue filling material containing adipose tissue-derived cells, and is produced by the following production method.

  As shown in FIG. 1, the method for manufacturing a biological tissue filling material according to this embodiment includes a digestion step S1 for digesting a biological tissue such as adipose tissue collected from a patient, and cells obtained by digesting the biological tissue. First centrifugation step S2 for centrifuging red blood cell components from the suspension, red blood cell removal step S3 for removing the separated red blood cell components, and tissue-derived cells and platelet components from the cell suspension from which the red blood cell components have been removed A second centrifugation step S4 for concentrating the supernatant, a supernatant removal step S5 for removing a portion of the centrifuged supernatant, and adding calcium chloride to the tissue-derived cells and platelet components after removing the supernatant And an adding step S6.

  In the digestion step S1, a living tissue collected from a patient is immersed in a digestive enzyme solution as it is without removing blood components and stirred for a predetermined time. Thereby, a biological tissue mass is decomposed | disassembled with a digestive enzyme, a tissue origin cell is isolated, and the cell suspension which floats in a digestive enzyme liquid is produced | generated. In addition, blood components contained in the living tissue also flow out into the digestive enzyme solution and are included in the cell suspension.

  In the first centrifugation step S2, the cell suspension is rotated at a relatively low centrifugal speed (for example, 900 rpm). By this first centrifugation step S2, the cell suspension is separated into the erythrocyte component 2 that settles in the centrifuge container 1 and the other component 3 that is disposed in the upper layer as shown in FIG. Is done. The upper layer contains blood components other than tissue-derived cells and red blood cells.

  In this state, the red blood cell component 2 is removed (step S3). In the removal step S3 of the erythrocyte component 2, only the upper layer may be sucked and transferred to another centrifuge container 1, or the syringe needle of the syringe is inserted near the bottom surface of the centrifuge container 1 to directly remove the erythrocyte component 2 by suction. You may decide.

  The second centrifugation step S4 rotates the cell suspension after the red blood cell component 2 is removed at a relatively high centrifugal speed (for example, 3600 rpm). By this second centrifugation step S4, as shown in FIG. 3, the cell suspension becomes a concentrated solution 4 of tissue-derived cells and platelet components that settles in the centrifuge container 1, and that disposed on the upper layer thereof. It is separated into other blood components 5.

  In this state, a part of the upper blood component 5 is removed to the position indicated by the broken line in FIG. 3 (step S5). In the supernatant removal step S5, only a part of the blood component 5 in the supernatant may be aspirated, or the syringe needle is inserted near the bottom of the centrifuge container 1 to derive the tissue concentrated in the lower layer. A part of the concentrated component 4 of the cells and platelet components and the blood component 5 of the supernatant (liquid in a range indicated by reference numeral 6 in FIG. 3) may be collected by suction. The blood component 5 of the supernatant contains thrombin.

  Then, in addition step S6, calcium chloride is added to the liquid 6 containing a part of the tissue-derived cell and platelet component concentrate 4 and the blood component 5 of the supernatant, whereby calcium chloride acts on thrombin in the liquid 6. It can be made to gel. Thereby, the biological tissue filling material which concerns on this embodiment can be manufactured.

  According to the manufacturing method according to the present embodiment, in the process of extracting tissue-derived cells from a biological tissue, a gel-like biological tissue filling material can be easily manufactured simply by adding calcium chloride using thrombin obtained at the same time. be able to. Conventionally, when tissue-derived cells are separated and extracted, thrombin has been discarded, so there is an advantage that it can be used without waste.

  In addition, since the tissue filling material manufactured by the above manufacturing method is formed in a gel shape, the fluidity is reduced, and it is easy to flow without filling from the filling position when the tissue filling portion is filled. To settle. As a result, the tissue-derived cells grow over time, so that the living tissue can be efficiently repaired.

  Moreover, since the biological tissue filling material produced | generated in the gel form has low fluidity | liquidity, there also exists an advantage that the handling is easy. In other words, in the case of a liquid with high fluidity, instruments for handling such as syringes and catheters are limited, but if it is a gel-like living tissue filling material, there is no such inconvenience and handling is easy. It is.

  Furthermore, since the gelatinous tissue replacement material produced in this manner contains abundant platelet components that promote the growth of tissue-derived cells, it promotes rapid growth of tissue-derived cells, There is an advantage that the organization can be repaired at an early stage.

In the present embodiment, the fatty tissue is exemplified as the living tissue. However, instead of this, any other living tissue may be used.
Moreover, when thrombin contained in the blood component is small, sufficient gelation is not performed by adding calcium chloride. In such a case, thrombin may be added.

It is a flowchart which shows the manufacturing method of the biological tissue filling material which concerns on one Embodiment of this invention. It is a figure which shows the cell suspension isolate | separated by the 1st centrifugation step in the manufacturing method of the biological tissue filling material of FIG. It is a figure which shows the cell suspension isolate | separated by the 2nd centrifugation step in the manufacturing method of the biological tissue filling material of FIG.

Explanation of symbols

2 Red blood cell component 5 Blood component (supernatant)
6 Liquid S2 First centrifugation step S3 Red blood cell removal step S4 Second centrifugation step S5 Supernatant removal step S6 Addition step

Claims (4)

  1.   The cell suspension obtained by digesting the biological tissue is centrifuged, the separated erythrocyte component is removed, the cell suspension after removal of the erythrocyte component is centrifuged again, and a part of the separated supernatant A gel-like biological tissue filling material containing tissue-derived cells produced by removing calcium and adding calcium chloride to a liquid containing tissue-derived cells and platelet components after removal of the supernatant.
  2.   The biological tissue filling material according to claim 1, wherein the biological tissue is an adipose tissue.
  3. A first centrifugation step of centrifuging red blood cell components from a cell suspension obtained by digesting biological tissue;
    An erythrocyte removal step for removing erythrocyte components separated by the first centrifugation step;
    A second centrifugation step of concentrating tissue-derived cells and platelet components from the cell suspension after removal of the red blood cell components by the red blood cell removal step;
    A supernatant removal step of removing a portion of the supernatant centrifuged by the second centrifugation step;
    A method for producing a biological tissue filling material, comprising: an addition step of adding calcium chloride to a liquid containing tissue-derived cells and platelet components after removal of the supernatant in the supernatant removal step.
  4.   The method of manufacturing a biological tissue filling material according to claim 3, wherein the biological tissue is an adipose tissue.
JP2007271575A 2007-10-18 2007-10-18 Biological tissue filling material and manufacturing method thereof Expired - Fee Related JP5094327B2 (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003320012A (en) * 2002-04-30 2003-11-11 Olympus Optical Co Ltd Method and device of manufacturing living tissue filling body and humor collecting container
JP2005160669A (en) * 2003-12-02 2005-06-23 Olympus Corp Manufacturing method of biological tissue prosthesis

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003320012A (en) * 2002-04-30 2003-11-11 Olympus Optical Co Ltd Method and device of manufacturing living tissue filling body and humor collecting container
JP2005160669A (en) * 2003-12-02 2005-06-23 Olympus Corp Manufacturing method of biological tissue prosthesis

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