WO1999012589A1 - Organe artificiel - Google Patents

Organe artificiel Download PDF

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Publication number
WO1999012589A1
WO1999012589A1 PCT/JP1998/004030 JP9804030W WO9912589A1 WO 1999012589 A1 WO1999012589 A1 WO 1999012589A1 JP 9804030 W JP9804030 W JP 9804030W WO 9912589 A1 WO9912589 A1 WO 9912589A1
Authority
WO
WIPO (PCT)
Prior art keywords
matrix
living cells
artificial
artificial organ
laminin
Prior art date
Application number
PCT/JP1998/004030
Other languages
English (en)
Japanese (ja)
Inventor
Masato Nagaki
Keizaburo Miki
Original Assignee
Foryou Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Foryou Corporation filed Critical Foryou Corporation
Publication of WO1999012589A1 publication Critical patent/WO1999012589A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3621—Extra-corporeal blood circuits
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3621—Extra-corporeal blood circuits
    • A61M1/3623—Means for actively controlling temperature of blood

Definitions

  • the present invention relates to an artificial organ using living cells such as hepatocytes, Teng cells, and kidney cells.
  • Known artificial organs in which living cells such as hepatocytes are dispersed and held in a matrix have been known.
  • Known artificial organs of this type contain living cells dispersed in a matrix inside a hollow fiber membrane, circulate a body fluid, and make the living cells perform their original physiological action, By returning to the patient again, it assists or substitutes for the organs of the patient whose function has been reduced or lost.
  • Known matrices include intercellular substances such as collagen and fibronectin, and resins such as poly-N-p-vinylbenzyl-D-lactamide (PVLA).
  • an object of the present invention is to provide an artificial organ using living cells that can be used for a long period of time while maintaining the function of the living cells at a high level.
  • the present inventors have conducted intensive studies and have found that by using laminin as a matrix, living cells are three-dimensionally retained in a natural form. As a result, the function of living cells is maintained at a high level for a long period of time. The present inventors have found that living cells can be kept alive throughout the present invention and completed the present invention.
  • the present invention provides a matrix containing 50% by weight or more of laminin, living cells dispersed in the matrix, and a storage body that stores the matrix and the living cells, and that allows liquid to flow into and out of the outside.
  • an artificial organ comprising:
  • the living cells are three-dimensionally retained in a natural form, and as a result, the function of the living cells is enhanced. Live cells can be kept alive for a long period of time while maintaining them.
  • FIG. 1 is a schematic view showing one example of a preferred embodiment of the artificial organ of the present invention.
  • FIG. 2 is a graph showing the relationship between the number of days after the start of blood treatment and the ratio of albumin mRNAZS-actin mRNA in hepatocytes in the artificial livers of the present invention and the comparative example.
  • FIG. 3 is a graph showing the relationship between the number of days after the start of blood treatment and the amount of albumin secreted by hepatocytes in the artificial livers of the present invention and the comparative example.
  • the living cells used in the artificial organ of the present invention are cells that have some physiological action on body fluids, and preferred examples include hepatocytes, mucous cells, and kidney cells, but are not limited thereto. Absent.
  • the origin of the living cells is not particularly limited, and may be any animal that exhibits the same performance as the internal organs of humans or humans. Preferred animals include pigeons, baboons, monkeys, hidges, dogs, dogs, and the like. Can be mentioned. These animals also include those that have been genetically engineered.
  • a matrix for retaining living cells As a matrix for retaining living cells, a matrix containing laminin at 50% by weight or more is used.
  • the content of laminin in the matrix is 50% by weight or more, and the total amount may be laminin.
  • a mixture of laminin and collagen IV is also preferred.
  • the weight ratio of laminin to collagen IV is preferably about 6: 4 to 8: 2.
  • the total amount of laminin and collagen IV is preferably at least 70% by weight, more preferably at least 90% by weight, particularly preferably at least 95% by weight of the whole matrix.
  • the matrix may contain substances other than laminin and collagen IV as long as the effects of the present invention are not significantly impaired.
  • Such substances include resins such as entactin, proteoglycan and PVLA which have been conventionally used as a matrix, and impurities which may accompany laminin / collagen IV.
  • the amount of these substances other than laminin and collagen IV does not significantly impair the effects of the present invention, and is usually 30% by weight or less, preferably 10% by weight or less, and more preferably 5% by weight or less.
  • Living cells are maintained in a dispersed state in the matrix. By using the above matrix, living cells are three-dimensionally maintained in a natural state close to the living body. Straightforward The vesicles can be kept alive. Viable cell density in the matrix and living cells in a mixture of is not particularly limited, but is usually 1 0 5 m I ⁇ 1 0 9 or Zm I about, preferably, 1 0 7 I ⁇ 1 0 8 pieces It is about Zm I. Also, the total number of viable cells, the type and the viable cells may be appropriately selected depending on the performance of the artificial organ to determine, for example, in the case of hepatocytes, 1 0 1 0 to 4 X 1 0 1 about 0 appropriate It is.
  • a hollow fiber membrane can be exemplified as a preferred form of the housing. That is, the inside of the hollow fiber membrane (that is, the hollow portion inside the fiber) or the outside is filled with a mixture of viable cells and a matrix.
  • the molecular weight cut off of the hollow fiber membrane is such that cells do not flow out and that the flow of bodily fluids is performed smoothly, that is, about tens of thousands to 100,000 daltons are preferable Particularly preferably, it is about 100,000 daltons.
  • the inner diameter of the hollow fiber membrane is not particularly limited, but is preferably about 250 to 350 m, and the length is also not particularly limited, but is usually about 100 to 350 mm.
  • the material of the hollow fiber membrane may be any material as long as it does not have a harmful effect on bodily fluids, and examples thereof include polysulfone and polyester, but are not limited thereto. Not something. It is preferable to bundle hundreds to thousands of such hollow fiber membranes filled with the live cell Z matrix mixture and store them in a cartridge having a liquid inlet and a liquid outlet. A bundle of hundreds or thousands of hollow fiber membranes having the above-described physical properties and contained in a cartridge having a liquid inlet and a liquid outlet is commercially available as an artificial dialysis device. In the present invention, such a commercially available artificial dialysis device can be preferably used as a container for an artificial organ.
  • the container is not limited to the hollow fiber membrane as described above, and may be, for example, a bag-shaped flat membrane or a form like an ordinary container. Anything that has a structure in which it can be housed, brought into contact with bodily fluids, and withdrawn bodily fluids after contact is included in the scope of the present invention.
  • the artificial organ of the present invention is obtained by mixing a solution of a substance constituting the matrix (hereinafter, sometimes referred to as a “matrix substance”) with a suspension of living cells, and filling the mixture into the container.
  • a solution of a substance constituting the matrix hereinafter, sometimes referred to as a “matrix substance”
  • a suspension of living cells It can be manufactured by the following.
  • Dissolution of matrix material As a solvent for the solution, any solvent can be used as long as it can dissolve the matrix substance and does not adversely affect the living cells. Preferred examples thereof include a culture solution, physiological saline, and a buffer solution. Can be mentioned.
  • the concentration of the matrix substance in the matrix substance solution is not particularly limited, but is preferably about 0.1 to 1.5 w / v%, particularly preferably about 1 w / V ⁇ 1 ⁇ 2.
  • the solvent for the suspension of the living cells those commonly used for suspending living cells, such as a physiological saline buffer, can be used.
  • Cell density of viable cells in suspension is particularly preferably but 1 0 5-1 0 about nine Zm l are not limited, and more preferably from 1 0 7 to 1 0 about eight Roh m I.
  • the mixing ratio between the matrix material solution and the viable cell suspension is selected to achieve the preferred viable cell density described above.
  • the container is a hollow fiber membrane
  • the mixture of the matrix substance solution and the live cell suspension is poured into the hollow part of the hollow fiber and left at 37 ° C to gel the matrix substance. Live cells dispersed in a gel matrix can be obtained.
  • Blood is drawn from the patient's femoral artery by the pump 10 and led to the plasma separator 12, where it is separated into blood cell components and plasma.
  • the liquid sending speed by the pump 10 is preferably 60 to 150 mI.
  • the plasma component is led to the heat exchanger 16 by the pump 14.
  • the pumping speed of the pump 14 is preferably about 30 mIZ.
  • the heat exchanger 16 is used to maintain the plasma temperature at the human body temperature (37 ° C).
  • the plasma is then led to an oxigenator 18 located adjacent to the heat exchanger 16 where it is supplied with oxygen. Oxygen supply is beneficial, but not essential, for the survival of living cells.
  • the plasma that has passed through the oxygenator 18 is guided to the artificial organ 20 of the present invention, and comes into contact with the living cells held in the matrix.
  • the artificial organ 20 shown in the figure is obtained by filling a hollow portion of a large number of hollow fiber membranes with a matrix matrix living cell mixture in a cartridge having a liquid inlet 22 and a liquid outlet 24.
  • the plasma is guided to the hollow portion inside the hollow fiber membrane to ensure sufficient contact with the living cells, and the side of the hollow fiber membrane is It is preferable that the processed plasma that has flowed out from the large number of holes in the liquid outlet is sent out from the liquid outlet 24.
  • the processed plasma sent out from the liquid outlet 24 is It merges with the blood cell component separated by the plasma separator 12 and is returned to the patient's body.
  • the mode of use of the artificial organ is not limited to the above, and for example, a type that is implanted in the body is also possible.
  • the artificial organ of the present invention can be used for assisting or substituting for the organ in a patient whose function has been reduced or lost.
  • the living cells are hepatocytes
  • the life of patients with severe liver diseases such as fulminant hepatitis and postoperative liver failure can be maintained by using the artificial organ of the present invention.
  • This commercially available dialysis machine converts 250 polysulfone hollow fiber membranes with a molecular weight cut off of 100,000 daltons, an inner diameter of 260 /, a wall thickness of 75 / im, and a length of 18 Omm into a cartridge with a diameter of 5 cm and a length of 20 cm. It is stored.
  • plastic polyurethane
  • collagen I collagen I
  • PVLA polyurethane
  • a 25 kg pig male, 27 weeks old was laparotomized, the hepatic artery was ligated, and the portal vein and the inferior vena cava were sutured. This was used as a liver failure model.
  • the femoral artery of this pig was connected to the extracorporeal blood circulation device shown in Fig. 1 (which was equipped with the artificial liver of Example 1 or Comparative Example 1, 2 or 3 as an artificial organ), and was described earlier with reference to Fig. 1. By the way, Blood processing was performed.
  • actin mRNA has the highest value in plate culture, and it can be used as an index of liver cell vitality by using this as the denominator to determine the ratio of albumin mRNA amount to the molecule. .
  • the hepatocytes maintained a high vitality even at the 9th word after the start of the blood treatment, whereas those defined by the present invention.
  • the vitality of hepatocytes was already low from 1 day after the start of blood treatment, and hardly functioned after 9 days.
  • Liver cells are derived from rat, except that the amount of the mixture of the matrix material solution and the hepatocyte suspension is a 25 m I (therefore, the total number of hepatocytes 2. 25 X 1 0 9 pieces), performed
  • An artificial liver was prepared in the same manner as in Example 1 (Example 2).
  • PVLA Comparative Example 4
  • collagen gel sandwich hepatocytes sandwiched between collagen gels
  • collagen gel Comparative Example 6
  • collagen An artificial liver was prepared in the same manner using a plastic coated with a commercially available plastic (polyurethane) (Comparative Example 7).
  • the prepared artificial liver was incorporated into the same extracorporeal blood circulation system as in Example 1, and blood treatment was performed using the same hepatic failure model pig as in Example 1.
  • blood treatment was performed using the same hepatic failure model pig as in Example 1.
  • the amount of albumin in the plasma after the treatment ie, the amount of albumin secreted by hepatocytes
  • Albumin secretion is an important function of hepatocytes.
  • the artificial liver of the present invention While the amount of albumin secreted by the vesicles was kept high after the start of blood treatment, the present invention also showed that the artificial liver of the comparative example had the highest level of comparative example 5 after the start of blood treatment on day 16 after the present invention. Less than half. Therefore, according to the present invention, it can be seen that hepatocytes can be maintained over a long period of time while maintaining the ability of hepatocytes at a high level.

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  • Health & Medical Sciences (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Vascular Medicine (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Anesthesiology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Cardiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • External Artificial Organs (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne un organe artificiel préparé à partir de cellules vivantes et pouvant être utilisé longtemps tout en conservant à un niveau élevé les fonctions des cellules vivantes. Cet organe artificiel comprend une matrice contenant au moins 50 % en poids de laminine, des cellules vivantes dispersées dans la matrice, ainsi qu'une enveloppe logeant la matrice et les cellules vivantes et permettant le transfert de liquide de ou vers l'extérieur.
PCT/JP1998/004030 1997-09-09 1998-09-09 Organe artificiel WO1999012589A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP9261045A JPH1176400A (ja) 1997-09-09 1997-09-09 人工臓器
JP9/261045 1997-09-09

Publications (1)

Publication Number Publication Date
WO1999012589A1 true WO1999012589A1 (fr) 1999-03-18

Family

ID=17356299

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1998/004030 WO1999012589A1 (fr) 1997-09-09 1998-09-09 Organe artificiel

Country Status (2)

Country Link
JP (1) JPH1176400A (fr)
WO (1) WO1999012589A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102210889A (zh) * 2010-04-02 2011-10-12 瑞德肝脏疾病研究(上海)有限公司 一种用于人工肝透析的体外生物材料及其制备方法

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2375505A1 (fr) * 1999-06-21 2000-12-28 The General Hospital Corporation Systemes de culture cellulaire et procedes pour dispositifs d'assistance aux organes
US8304238B2 (en) * 2003-03-24 2012-11-06 Nat'l Institute for Environmental Studies Cell culture medium and immobilized preparation of cell adhesion protein or peptide
MX350338B (es) 2005-08-26 2017-09-04 Univ Minnesota Descelularizacion y recelularizacion de organos y tejidos.
US20120183944A1 (en) * 2009-03-31 2012-07-19 Regents Of The Univeristy Of Minnesota Decellularization and recellularization of organs and tissues
SG188369A1 (en) 2010-09-01 2013-04-30 Univ Minnesota Methods of recellularizing a tissue or organ for improved transplantability
WO2014151739A1 (fr) 2013-03-15 2014-09-25 Miromatrix Medical Inc. Utilisation de foie décellularisé par perfusion pour la recellularisation de cellules d'îlot de langerhans
EP3509651B1 (fr) 2016-09-06 2022-12-28 Miromatrix Medical Inc. Utilisation de sérum de foie réséqué pour ingénierie du foie entier

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6417653A (en) * 1987-07-13 1989-01-20 Otsuka Pharma Co Ltd Laminate type hybrid artificial liver apparatus
JPH04322657A (ja) * 1991-02-15 1992-11-12 Tokyo Univ 生体細胞の成長並びに機能分化の促進・制御方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6417653A (en) * 1987-07-13 1989-01-20 Otsuka Pharma Co Ltd Laminate type hybrid artificial liver apparatus
JPH04322657A (ja) * 1991-02-15 1992-11-12 Tokyo Univ 生体細胞の成長並びに機能分化の促進・制御方法

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102210889A (zh) * 2010-04-02 2011-10-12 瑞德肝脏疾病研究(上海)有限公司 一种用于人工肝透析的体外生物材料及其制备方法

Also Published As

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