Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Solid support and method of culturing cells on said solid...
Reexamination Certificate
2001-03-02
2002-08-13
Lankford, Jr., Leon B. (Department: 1651)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Solid support and method of culturing cells on said solid...
C435S383000
Reexamination Certificate
active
06432713
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The invention relates to a culture technology of a cell or tissue employed by a tissue engineering that is applied to a cell or tissue engineering or genetic treatment, particularly to a method of and an apparatus for cultivating a cell or tissue (hereinafter referred to as culture method and culture apparatus) for use in an in vitro culture of a cell or tissue that is needed for restoring a damaged tissue of human body.
2. Description of the Related Art
There are following methods for restoring a damaged tissue or a pathogenic part of a living body. The first method is to substitute the damaged tissue or pathogenic part for materials other than a living body such as plastic, metal, ceramic as restoring means of the damaged tissue or the pathogenic part. As substitutable materials, there are ceramics and stainless steel for bones, a polyethylene resin for joints, and a vinyl resin for blood vessels. Second method is to substitute the damaged tissue or pathogenic part for parts of other animals or the different position of the living body. As the substitutable tissue in the second method, there are, for example, skins. The third method is to transplant of internal organs of other people.
In the first method, there is a drawback that the materials other than the living body such as plastic, metal, ceramic need to be substituted periodically by others when they are worn or consumed or materials separated from the materials other than the living body by the wear thereof affects adversely on the living body. Further, in a blood vessel made of a synthetic polymer, there is a report that an interior of the blood vessel is clogged when it is used for a long period of time. In the third method, if there is no donor for supplies his or her internal organs to be transplanted, it is impossible to carry out the third method. Even if the third method is carried out, there still remains a problem of immunological rejection between internal organs of two people.
Accordingly, a method of restoring a damaged tissue or a pathogenic part of a living body that is expected to be carried out is to substitute the damaged part of a cell or tissue by a cell or tissue that is obtained by cultivating a cell or tissue in vivo or in vitro. It is reported in current researches that there is a possibility in many tissues such as skins, cartilage, bones, blood vessels, livers, and pancreas. If a cell or tissue derived from a living body is cultivated inside or outside the living body of a patient, and the cell or tissue obtained by the culture is applied to the restoration of a damaged part, a tissue can be regenerated in the body, and further the tissue applied to the restoration would have gene of the patient per se, there does not occur immunological rejection, and further, a chemical substance such as synthetic polymer other than a living material does not adversely affect a living body, thereby realizing an ideal treatment.
There has been proposed and disclosed as a technology of this type in Japanese Patent Laid-Open Publication No. 9-313166 entitled “DEVICE FOR CULTURING CELL”. This technology needs to disassemble into each part every culture, to clean, to sterilize, and then reassemble the apparatus, resulting in a risk of contamination by bacteria after sterilization. Although each part of the apparatus can be assembled for preventing contamination by bacteria so as to perform a sterilization treatment by an autoclave (absolute pressure 2 atm. at 121° C.), this technology can not be employed in view of the contamination by bacteria because a pump and a pressure sensor include many electronic devices, a specific resin and oil. Accordingly, parts of the pump and pressure sensor are disassembled while only a passage through which a culture medium passes is taken out and is sterilized by chemicals, and other parts are sterilized by the autoclave, thereafter the pump and pressure sensor are assembled together with the apparatus, resulting in much labor and the increase of risk of contamination by various bacteria. Further, in the culture using an incubator (culture vessel), a pump or a controller is subjected to an adverse affection by a temperature or humidity, and also all the devices can not be accommodated in the incubator having a limited capacity. Accordingly, it is necessary to assemble the culture apparatus in a state where the incubator communicates with an open air for allowing piping, a power supply and a controlling electric wire to pass through a through hole of the incubator. Still further, since a pressure is applied to an entire circuit of a culture medium, the entire culture apparatus including parts of the pump and piping shall have a pressure resistant construction. As a result, it is very difficult to place the apparatus at high pressure e.g. not less than 1 MPa, and even if a high pressure is applied to the apparatus, the apparatus shall be high pressure resistant as a whole, resulting in a problem of high cost.
More still further, there is a research reported by Dr. Shuichi MIZUNO et al. in Harvard Medical School that a tissue of a living body is cultivated by applying a pressure to the living body as physical stimulation (see Materials Science and Engineering C6 (1998) 301-306). According to this research, a culture apparatus is formed as illustrated in FIG.
26
. Each constituent and function thereof in this culture apparatus is described now.
A pump
400
has a role to circulate a culture medium
402
therein and to pressurize the interior of a culture chamber
404
to supply a hydraulic pressure to a cell
406
or tissue, and it is formed of a pump for use in a liquid chromatograph, and further it has a control unit built therein for flowing a given amount of fluid.
A back pressure regulator
408
allows a pressure to escape through a valve
410
by opening the valve
410
when a pressure exceeds a pressure to be applied to the cell
406
or tissue exceeds so as to hold the pressure inside the culture chamber
404
constant. The back pressure regulator
408
is selectively provided in a circuit
426
, described later, depending on a pressure to be applied to the cell
406
.
The culture chamber
404
forms a space for cultivating the cell
406
or tissue, and a scaffold
412
formed of a sponge made of a collagen in which the cell
406
or tissue is transplanted is accommodated in the space. The cell
406
or tissue grows on the scaffold
412
formed of a sponge made of a collagen. A pressure sensor
414
detects a pressure inside the culture chamber
404
while a pressure monitor
416
indicates the pressure detected by the pressure sensor
414
. The pump
400
is controlled by the pressure detected by the pressure sensor
414
and it stops its operation when the detected pressure increases to a large extent.
A culture medium tank
418
stores therein the culture medium
402
adapted for the cell
406
or tissue to be cultivated and the culture medium
402
comprises e.g., amino acids, saccharides, salts, and so forth. The culture medium tank
418
communicates with an open air through a vent tube
422
that penetrates a closed stopper
420
, and a vent filter
424
prevents the culture medium
402
from being contaminated by an open air.
The culture apparatus is accommodated in an incubator forming a hermetically sealed space. The incubator is a space for forming a pleasant cultivating atmosphere and it is maintained under the optimum temperature, humidity and gas concentration (oxygen and carbon dioxide) that is optimized for the cell or tissue. The culture medium
402
is filled in the circuit
426
by the pump
400
and circulated therein. The oxygen and carbon dioxide are soluble in the culture medium
402
after they pass through the vent filter
424
, and the culture medium
402
is kept under appropriate oxygen concentration and carbon dioxide concentration. When the pump
400
is operated, a pressure inside the culture chamber
404
gradually increases. When the pressure exceeds a given value set by the back pressure
Glowacki Julie
Kinouchi Ibuki
Mizuno Shuichi
Takagi Takao
Takai Hidetada
Connolly Bove & Lodge & Hutz LLP
Lankford , Jr. Leon B.
Takagi Industrial Co., Ltd.
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