Cultured Cell Sheet, Production Method and Tissue Repair Method Using Thereof

a production method and cell sheet technology, applied in the field of cultured cell sheet, a production method, and an applied tissue repair method thereof, can solve the problems of biological toxicity, limited site to be applied, toxicity to living organisms, etc., and achieve excellent flexibility, suppress air leakage, blood leakage or bodily fluid leakage, and high adhesiveness.

Inactive Publication Date: 2008-06-05
CELLSEED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0012]The highly adhesive cultured cell sheet obtained by the present invention has extremely high adhesiveness to a leaking site of the surface of an organ and an excellent flexibility. Therefore, the use of the cell sheet of the present invention enables to suppress air leakage, blood leakage or bodily fluid leakage from the surface of an organ. Consequently, the present invention is extremely useful in biological and medical fields, etc., such as cell engineering and medical engineering.

Problems solved by technology

However, cyanoacrylate adhesive agents are synthetic agents which are not originally endogenous, and prevent healing by producing formaldehyde by hydrolyzation of a cured monomer, which causes toxicity to living organisms.
Therefore, the problems exist that the site to be applied was limited, and the adhesive agents should not be applied to a site near a central nerve or blood vessel.
Although the gelatin-aldehyde adhesive agents also have sufficiently high adhesive intensity, since toxic aldehyde compounds are employed as a cross-linking agent, they are also biologically toxic.
Although this type of adhesive agent is less toxic than the above described synthetic adhesive agents, the adhesiveness is low, and large amounts of fibrin glue should be used, since the fibrin glue itself is metabolized in vivo.
Further, recently, some problems are being indicated relating to the topical inflammation at the site where the adhesive agent has been applied, since the fibrin glue is prepared and purified heterogeneously or the mechanisms of the adhesiveness of the fibrin glue are the same as the blood coagulation reaction.
However, in cases where the cells are harvested by the above-mentioned chemical agent treatment, some disadvantages have been pointed out: the treatment method is cumbersome and complicated; the potential for contamination by impurities is increasing; and examples of defects, in which cells are caused to degenerate or are damaged by the chemical treatment, and lose their original function.

Method used

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  • Cultured Cell Sheet, Production Method and Tissue Repair Method Using Thereof
  • Cultured Cell Sheet, Production Method and Tissue Repair Method Using Thereof
  • Cultured Cell Sheet, Production Method and Tissue Repair Method Using Thereof

Examples

Experimental program
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examples

[0055]Hereinafter, the present invention will be explained in further detail based on the following Examples, which are not intended to limit the scope of the present invention in any way.

examples 1 and 2

[0056]To a commercial culture dish with a diameter of 3.5 cm (Falcon 3001, manufactured by Becton Dickinson Labware), 0.07 ml of the solution of N-isopropyl acrylamide monomer dissolved in isopropyl alcohol at a concentration of 53% (Example 1) or 54% (Example 2) was applied. The culture dish was exposed to electron beams at an intensity of 0.25 MGy, and the N-isopropyl acrylamide polymer (PIPAAm) was immobilized on a surface of the culture dish. After irradiation, the culture dish was washed with ion-exchanged water to remove a residual monomer and the PIPAAm that did not bind to the culture dish, was then dried inside a clean bench, and sterilized by ethylene oxide gas, to obtain a cell culture support material coated with a temperature responsive polymer.

[0057]The amount of the temperature responsive polymer on the support surface was measured. As a result, it was found that the supports' surface was coated with the temperature responsive polymer in an amount of 1.7 μg / cm2 (Examp...

example 3

[0061]The study of this example is conducted in a similar manner to that of Example 2, except that 250 μM β-aminopropylnitrile was added to the culture medium, when cell passage was started on the cell culture support material, 3 days after the start of culture in Example 2. The cultured cell sheet obtained in the presence of β-aminopropylnitrile was mechanically flexible. The air leakage site was closed, and contracture of the covered portion of the cultured cell sheet was not found. Accordingly, the usefulness of a cultured cell sheet having flexibility as a tissue repair material could be confirmed.

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Abstract

It is intended to provide a cultured cell sheet with excellent tissue adherence and flexibility. The above object can be achieved by culturing cells on a support for cell culture in which a surface of a substrate is coated with a temperature-responsive polymer whose lower or upper critical solution temperature against water is in the range of 0 and 80° C. along with a surfactant protein or a crosslinking inhibitor and producing a cultured cell sheet by detaching it by setting the temperature of the culture to the upper critical solution temperature or higher or to the lower critical solution temperature or lower.

Description

TECHNICAL FIELD[0001]The present invention relates to a cultured cell sheet, a production method, and an applied tissue repair method thereof, in medical and biological fields, etc.BACKGROUND ART[0002]Japan is becoming an aging society with the highest average life expectancy in the world. People are beginning to place more emphasis on living better, i.e., “the quality of life (QOL)”, rather than merely prolonging life. Under such circumstances, medical techniques are rapidly progressing, and techniques for reconstruction of organs damaged by trauma or disease are also advancing remarkably. Therefore, in recent years, a lot of attention has been focused on regenerative medical techniques in which organ tissue is reconstructed in the cell culture system employing cultured cells, and then transplanted to the target site.[0003]To carry out such treatments, it is necessary to use tissue adhesive agents for tissue adhesion. The tissue adhesive agents currently used in the clinical field ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/06A61F2/00A61L27/00A61L27/22A61L27/38A61L27/40A61L27/50A61L31/04A61L31/12A61L31/14
CPCA61L27/34A61L27/3804A61L27/3839C12N5/0688C12N2533/30A61L27/3895C12N5/0068A61P1/16A61P11/00A61P9/00
Inventor KANZAKI, MASATOYAMATO, MASAYUKIKOHNO, TINATSUSEKINE, HIDEKAZUKIKUCHI, AKIHIKOOKANO, TERUOONUKI, TAKAMASA
Owner CELLSEED
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