Title: Capsules containing
vital cells or tissues
United States Patent: 6,982,095
Issued: January 3, 2006
Inventors: Asada; Masanori (Osaka, JP);
Hatano; Yumi (Osaka, JP); Kamaguchi; Ryosei (Osaka, JP); Sunohara; Hideki
Assignee: Morishita Jintan Co., Ltd.
Appl. No.: 480720
Filed: June 28, 2001
PCT Filed: June 28, 2001
PCT NO: PCT/JP01/05606
371 Date: December 12, 2003
102(e) Date: December 12, 2003
PCT PUB.NO.: WO03/001927
PCT PUB. Date: January 9, 2003
Training Courses -- Pharm/Biotech/etc.
Capsules which contain vital cells or
tissues as the contents and in which these cells or tissues can be grown.
Since these cells or tissues can be grown in the capsules, an extremely
high cell density can be achieved. By using these capsules, foods having
an excellent effect of ameliorating intestinal disorders, etc. can be
provided. Moreover, it is possible to provide artificial seeds showing an
excellent storage stability and a high germination ratio which comprise
seamless soft capsules consisting of an innermost layer containing
indefinite embryo, indefinite shoots, multiple shoots, shoot apexes,
growing points, protocorm-like bodies, indefinite roots, hairy roots,
etc., an inner coating layer comprising a hardened oil and an outer
coating layer made of a biodegradable film comprising gelatin,
Description of the Invention
This application is the National Stage if
International Application No. PCT/JP01/05606, filed Jun. 28, 2001.
The present invention relates to capsules
including living (vital) cells or tissues and applications thereof. More
specifically, the present invention relates to capsules that include
living cells or tissues derived from microorganisms, plants or animals as
the inclusion and in which these cells or tissues can be grown. The
present invention also relates to foods containing such capsules and
artificial seed capsules having excellent storage stability and containing
redifferentiable plant cell tissues that germinate rapidly when being sown
Fermented foods obtained by allowing
microorganisms to act on foods have been eaten since old times. Yogurt,
which is a typical fermented food, is obtained by allowing lactic
bacteria, bifidobacteria or the like (hereinafter, unless otherwise
indicated, they are collectively abbreviated as "lactic bacteria") to act
on milk. When yogurt is ingested, lactic bacteria are delivered to the
intestines, act actively in the intestines and actively improve intestinal
disorders. However, since most of lactic bacteria die in the stomach due
to their strong acidity, only a few living lactic bacteria can reach the
In order to solve this problem, it has been attempted to include lactic
bacteria in an enteric capsule to deliver the lactic bacteria to the
intestines (for example, Japanese Laid-Open Patent Publication No.
8-242763). However, the lactic bacteria included in the capsule are
freeze-dried and it takes a long time until the freeze-dried lactic
bacteria absorb water and exhibit their activities again.
If living cells can be delivered to the intestines, the living cell can
exhibit their activities immediately and sufficiently. Therefore, there is
a demand for a technique for delivering living cells to the intestines.
On the other hand, the remarkable advance of biotechnology has affected
various fields such as development of pharmaceuticals or improvement of
plants. In the field of plants, there have been attempts to produce
artificial seeds from redifferentiable plant cell tissues (hereinafter,
referred to simply as "cell tissues"). In general, to produce artificial
seeds, at first, cell tissues are dispersed or suspended in a solution of
polysaccharides, low molecular weight of polymeric substance or
crosslinkable polymer. Then this dispersion or suspension is gelled so
that the cell tissues are enclosed in gels, followed by being molded into
a form of a bead, a plate, a bar or a fiber.
However, the artificial seeds in which cell tissues are enclosed in gels
as described above have no tolerance with respect to dryness when left at
a room temperature. Therefore, such artificial seeds have to be sown
within four days after they are prepared. In addition, these artificial
seeds cannot be stored even for one month, unless they are not stored in a
refrigerator or a liquid in order to prevent them from being dried. Thus
it is difficult to store them in a dry place, such as a barn of a farmer
or a storehouse, without a special refrigerating facility.
In order to improve this dryness problem, it has been attempted to cover
the surface of the gels with paraffin, wax or the like. It is possible to
improve the storage stability by covering the surface of the gels.
However, since paraffin or wax is stable and is hardly degraded, the
artificial seeds covered with paraffin or wax cannot germinate when they
are sown in soil. Therefore, it is necessary to make an opening for each
artificial seed when sowing, which is inconvenient. Thus such artificial
seeds cannot be used practically.
As described above, there is a demand for a technique to enclose living
cells and grow the living cells. Such technique can be applied not only in
a field of food chemistry, but also to a wide range of fields, for
example, pharmaceutical or agricultural field. In particular, in the field
of the artificial seeds, there is a demand for artificial seeds that can
be stored for at least three months in a dry place without a special
refrigerating facility, and can germinate in a few days when being sown in
soil. If such artificial seeds can be obtained, this technique can
actually be applied to a wide range of fields, for example, agriculture,
forestry, horticulture and floriculture, in combination with a technique
for cultivating plant cell tissues, a cloning technique or a virus-free
The inventors of the present invention
conducted in-depth research in order to solve the above problem.
Consequently the present inventors found that a capsule that includes
living cells or tissues suspended in a liquid can be produced, and that
this can be applied to the fields of foods and agriculture and thus
achieved the present invention.
The present invention provides a capsule including a liquid in which
living cell or tissue is suspended, wherein the cell or tissue can grow in
In a preferable embodiment, the capsule is a seamless soft capsule.
In one preferable embodiment, the cell or the tissue is a cell or a plant
tissue used for food.
In one preferable embodiment, the cell or the tissue is one or two or more
cells or tissues selected from the group consisting of lactic acid
bacteria (including bifidobacteria), Bacillus natto, baker's
yeasts, brewer's yeasts, filamentous fungus for brewing, single cell
algae, multicellular algae, edible plants, edible plant tissues and their
freeze-dried bacteria or tissues.
The present invention also is directed to food containing the
above-described capsule, and preferably the food containing the capsule is
fruit juice beverages, vegetable juice, health drinks, processed milk,
soybean milk, jelly, yogurt, lactic acid bacteria beverages, fermented
milk, carbonated drinks, near water, and pudding.
Furthermore, the present invention is directed to an artificial seed
including a redifferentiable plant cell tissue suspended in a liquid,
wherein the tissue is preserved in the liquid.
Furthermore, the present invention is directed to an artificial seed
formed of a seamless soft capsule having a structure of three or more
layers comprising an innermost layer, an inner layer covering the
innermost layer, and an outer layer covering the inner layer, wherein the
innermost layer includes a redifferentiable plant cell tissue, wherein the
inner layer is formed by an inner membrane comprising hardened oil as a
main component, and wherein the outer layer is formed of a biodegradable
In one preferable embodiment, the outer membrane is dried.
In one preferable embodiment, the redifferentiable plant cell tissue is
selected from the group consisting of adventitious embryos, adventitious
buds, multiple shoots, shoot apices, growing points, protocorm-like
bodies, adventitious roots, and capillary roots.
In one preferable embodiment, the inner membrane is microbiologically
degradable hardened oil that is solid at room temperature.
In another preferable embodiment, the outer membrane is a
microbiologically degradable outer membrane selected from the group
consisting of protein, polysaccharides, and biodegradable plastics.
BEST MODE FOR
CARRYING OUT THE INVENTION
Since it was difficult to capsulate
aqueous substances, only dry cells were capsulated. It was thought to be
impossible to grow the cells or the tissues that is enclosed in a capsule,
and therefore, no attempts were made to include and grow the cells or
tissues in a capsule. However, the inventors of the present invention
succeeded in capsulating aqueous substances for the first time, and found
that the cells or the tissues do not die and can grow even if they are
enclosed in the aqueous substance in the capsule, and thus achieved the
The present invention can provide the following effect, for example.
Conventionally, only freeze-dried lactic bacteria, bifidobacteria or the
like could be delivered, so that only a moderate effect of improving
intestinal disorders was obtained. On the contrary, by the present
invention, an immediate and potent effect of improving intestinal
disorders can be brought about by delivering living lactic bacteria,
bifidobacteria or the like. Furthermore, since cells or tissues can grow
in the capsules, capsules having a higher bacterium density (e.g., about
1010 to 1011/g of the capsule) can be obtained.
Therefore, the present invention can be used in a wider range of
applications such as bioreactors, pharmaceuticals, medicine, and
artificial seeds, in addition to food application as described above.
In a preferable embodiment, the ratio of the liquid enclosed in the
capsule (inner liquid) is about 30% by weight with respect to the total
weight of the capsule. In this case, the bacterial cell density of the
inner liquid becomes as high as 3.3×1011/ml. Unless the
bacterial cells are not included in the capsule, it is not possible to
grow the bacterial cells to such a high density. Furthermore, when the
bacterial cells are cultured and concentrated to such a high density, the
viscosity of the bacterial cell suspension becomes very high. Therefore,
it is very difficult to include such a high density of bacterial cell
suspension into the capsule from the outside with current techniques,
which proves the utility of the present invention more clear.
"Capsule" in the present specification refers to a structure that has an
outer membrane and is able to enclose or include a liquid, a suspension or
a culture solution containing cells or tissues within the outer membrane.
The capsule generally has a spherical shape, but is not limited to the
As the outer membrane of the capsule, any membrane can be used, as long as
it is conventionally used for capsules. As a membrane, membrane made of
natural polymer or a synthetic polymer can be used. When it is used for
food applications, a membrane made of natural polymer is preferable.
For the shape of the capsule, a two-layered structure can be used, and a
three-layered structure or a structure having more layers can be used. A
three-layered structure is preferable.
In the case of a three-layered structure, an innermost layer is a liquid
containing living cells or tissues, an intermediate layer that covers the
liquid of the innermost layer is a lipophilic membrane, and an outermost
layer is an outer membrane. It is preferable that the outer membrane has
one or more properties of edibility, easy to decompose (e.g.,
biodegradable), enteric properties, water-insolubility, and
biocompatibility according to the application.
The innermost layer (inclusion) is a liquid such as a water, a
physiological saline solution, a buffer, a culture medium and contains
components necessary to grow or maintain the lives of the cells or
Hereinafter, at first, capsules used in food applications will be
described, and then artificial seeds will be described.
For use in food applications, various oils and fats, fatty acids or fatty
acid ester of saccharides that are edible can be used as a lipophilic
membrane of the intermediate layer. Preferably, animal oils and fats,
vegetable oils and fats, biologically or chemically treated oils and fats
thereof can be used. Examples of edible oils and fats include, but are not
limited to, various deep frying oils, salad oil, hardened oil having a
melting point of 35° C. or less, vitamin E, wheat germ oil, sesame oil,
cocoa butter, butter, margarine, shortening, and fatty acid ester of
sucrose. By using these lipophilic materials, capsules enclosing aqueous
substances can be obtained.
When the capsule of the present invention is used for food applications,
as the edible outer membrane, natural polymer membrane such as gelatin,
agar, pectin, alginic acid, carrageenan, curdlan, starch, gellant gum,
glucomannan, or mixtures thereof can be used. Further, if desired, an
edible polymer membrane obtained by adding a protein, a glycoprotein, a
mucopolysaccharide, a saccharide, a sugar alcohol, a polyalcohol or the
like to the above edible outer membrane is also preferable as the outer
membrane. Specific examples of natural polymer include, but are not
limited to, Arabian gum, pullulan, dextran, xanthan gum, Locust bean gum,
collagen, and casein.
Furthermore, outer membranes that are soluble in the mouth, the stomach,
the small intestine or the large intestine or outer membranes that are
insoluble and discharged can be selected, if desired. The outer membrane
that is soluble in the intestines, for example, can be obtained by
combining gelatin or agar and pectin. For the outer membrane, glycerin can
be added in view of the moldability.
In the case of a three-layered structure, examples of preferable
combinations are shown, in the order of the intermediate layer (inner
membrane)—the outermost layer, for example, vitamin E oil—agar, wheat germ
oil—sodium alginate, a fatty acid ester of sucrose—gelatin, medium fatty
acid ester of glycerin—carrageenan. Among these, vitamin E oil—2 to 4%
agar and wheat germ oil—2 to 4% sodium alginate are preferable in term of
the properties of the obtained capsule.
The constitution ratio of the inclusion (the innermost layer), the
intermediate layer and the outermost layer depends on the size of the
capsule, but is preferably 10 to 70: 10 to 50: 5 to 50 by weight, and more
preferably 30 to 50: 25 to 40: 25 to 40.
The capsule of the present invention is preferably a seamless soft
capsule. If the seamless capsule has a multi-layered structure, each of
the intermediate layer and the outermost layer can be made thin. Thus, it
seems that a substance that is necessary for growth of the cells or the
tissues can be more easily moved between the inside and the outside of the
capsule, thereby enabling the cells or the tissues in the capsule to grow.
Furthermore, the grown cells cannot be leaked to the outside of the
Furthermore, depending on the application, the outermost layer can be a
membrane having a large pore size, good permeability and a low mass
transfer resistance, or a membrane having a small pore size, high barrier
properties and a high mass transfer resistance. Moreover, a capsule having
various functions can be prepared, for example, by adding a charged
substance to the outer membrane so that the outer membrane have a
selectivity for permeating a charged material.
There is no limitation regarding the cells or the tissues to be included
in the capsule. Bacteria, yeasts, molds, algae, plant cells, plant or
animal tissues can be used, depending on the application.
Examples of cells or plant tissues used in food applications include
lactic bacteria (including bifidobacteria), Bacillus natto, baker's
yeasts, brewer's yeasts (e.g., wine yeasts, sake yeasts, soybean yeasts,
or soy sauce yeasts), filamentous fungus for brewing (e.g., Aspergillus
oryzae), single cell algae (e.g., Chlorella pyrenoidosa, Spirulina),
multicellular algae (e.g., Undaria pinnatifida, tangle), edible
plants and edible plant tissues (e.g., ginseng).
The capsules including these cells or tissues can be produced by a method
commonly used by those skilled in the art. When the capsule has a
three-layered structure, it is most preferable to make a seamless soft
capsule. A method for producing a seamless soft capsule is disclosed in,
for example, Japanese Laid-Open Patent Publication No. 5-31352, Food
Processing Technology vol. 15, pp. 28-33, 1995, or Bioscience and
Industry, vol. 58, No. 7, pp. 31-34 (2000). In particular, it is more
preferable to produce a seamless soft capsule with an oily substance
(e.g., hardened oil) as the intermediate layer (inner membrane) using a
dropping method with a triple tube nozzle.
The amount of the cells or tissues included in the capsule can range from
the minimum amount that allows survival or growth to a cell density that
can be achieved by culturing and collecting cells or tissues by filtration
or centrifugation. However, in the present invention, since it is possible
to increase the number of cells or tissues in the capsule to a high
density, which is a feature of the present invention, the concentration of
the cells or tissues to be inoculated in the capsule is sufficient as low
as that of sub-cultivation level.
The average particle size of the seamless soft capsule depends on the
application of the capsule. In general, the mean particle size is 0.1 mm
to 10 mm, more preferably 0.2 to 8 mm. When the capsule is used for foods,
the average particle size is preferably 4 mm or less, and more preferably
0.1 mm to 2 mm. Such a seamless soft capsule can be drunk smoothly as it
is, however, in order to eat more smoothly, the surface of the capsule can
be coated with a paste or a thickener such as starch, a hydrolyzed starch
The living cells or tissues included in the seamless soft capsule can grow
even in the capsule preferably by suspending the capsule in a suitable
cultivation medium. In particular, in the case where a membrane material
that requires multivalent metal ions for gel formation, such as alginate,
is used as an outer membrane, it is preferable to add multivalent metal
ions to the medium in a concentration necessary to maintain the gel
intensity. Therefore, in the case of alginic acid, calcium chloride,
strontium chloride, barium chloride, or aluminum chloride can be added in
0.01 to 5% by weight with respect to the weight of the medium, preferably
0.5 to 3% by weight.
For example, when a capsule including bifidobacteria is cultured, the
bifidobacteria can be cultured and grown in the capsule, so that the
number of the living bifidobacteria in the capsule is as large as several
tens billion/g or more of the capsule. In addition, since living cells are
cultured in the capsule, metabolites (e.g., bacteriocin, polysaccharides),
are kept in the capsule, while such metabolites may be lost when bacteria
is cultured in a tank and washed with water. Therefore, when
bifidobacteria that has grown in the capsule is ingested in the form of
the whole capsule, a more immediate and potent effect of improving
intestinal disorders can be exhibited compared with the ingestion of a
powder of freeze-dried bacteria. Furthermore, an effect of making the skin
and the articulation smooth also can be provided.
In the capsule including living lactic bacteria, the lactic bacteria can
be cultured and grown in the capsule. Therefore, when the capsule is used
as a bioreactor of lactic acid fermentation, lactic acid can be produced
with high efficiency, because there is not any contamination of lactic
bacteria, and is easy to recover the lactic acid.
In the present invention, the cells or the tissues included in the capsule
can grow in a desired medium. The capsule including the grown cells or
tissues can be used for foods or the like. For example, lactic acid
bacteria (bifidobacteria), ginseng tissues or the like are enclosed
in a capsule and are grown in the capsule. After then, the capsules are
added to fruit juice beverages, vegetable juice, health drinks, soybean
milk, jelly, processed milk, yogurt, lactic acid bacteria beverages,
fermented milk, carbonated drinks, near water, pudding, or the like, so as
to produce foods containing the capsules of the present invention. In the
present invention, "jelly" includes foods in a gel-type. The foods to
which the capsules can be added are not limited to the above foods.
There is no particular limitation regarding the amount of the capsules
added to foods, but 0.1 to 10 g with respect to 100 g of food is
preferable, and 0.5 to 3 g is more preferable.
Next, the artificial seeds of the present invention will be described. The
artificial seeds of the present invention can preferably be made into
seamless soft capsules having a three-layered structure having the
innermost layer, the inner layer covering the innermost layer and the
outer layer covering the inner layer or a multi-layered structure having
more than three layers. FIG. 1 shows a schematic cross-sectional view of
the artificial seeds of the present invention. FIG. 1 is a diagram for the
case of the three-layered structure. The innermost layer includes, for
example, an adventitious embryo, which is a cell tissue. This innermost
layer is filled with a liquid or gel that contains components necessary to
maintain the life of the adventitious embryo and growth factor thereof.
The inner layer is composed of an inner membrane (shown as "inner membrane
(hardened oil membrane)" in FIG. 1) having hardened oil as the main
component. This hardened oil membrane prevents water of the inclusion from
evaporating, and suppresses oxygen permeation. The outermost layer is
shown as "outer membrane (gelatin membrane)" in FIG. 1, and retains the
physical strength and suppresses oxygen permeation.
In the description of various academic articles or patent documents, a
single bead of calcium alginate gel may be referred to as "capsule".
However, such a conventional capsule is completely different from the
artificial seed of the present invention, as evident from the production
method described later. Furthermore, the surface of the artificial seed
capsules of the present invention is preferably dried, which shows that
the artificial seed capsules of the present invention are different from
the conventional capsule composed of wet alginic acid-gel.
As a cell tissue used in the present invention and included in the
innermost layer, adventitious embryos, adventitious buds, multiple shoots,
shoot apices, growing points, protocorm-like bodies, adventitious roots,
capillary roots or the like can preferably be used. Virus-free tissues can
be used as well.
In order to maintain the lives of cell tissues, it is preferable to
suspend the cell tissues in, for example, water, a physiological saline, a
buffer, a culture medium, or a liquid or a gel that contains necessary
components to maintain an activity of differentiation (germination) of the
cell tissues (hereinafter, collectively referred to as "medium"). A medium
that is suitable for a desired plant and used by those skilled in the art
are, for example, the basal medium of Murashige & Skoog (1962:
hereinafter, referred to as "MS medium") or a modified medium thereof,
although there is no limitation thereto. Furthermore, plant hormone,
coconut milk, casein hydrolysate, yeast extract and the like, which are
usually used in a culture medium, also can be added. In addition, in the
case where it takes a long time to decompose the inner membrane and to
germinate from the tissue after sowing, there is a possibility that a
damage to the tissue is caused by microorganisms. In such a case,
antibacterial agents can be added in order to suppress the growth of the
As the inner membrane, oil and fat containing hardened oil as the main
component can preferably be used. "Containing hardened oil as the main
component" means that the inner membrane is composed of only hardened oil
or a mixture of hardened oil and other oil and fats so as to adjust the
desired properties. A preferable inner membrane is a microbiologically
degradable hardened oil that is solid at room temperature. "Hardened oil
that is solid at room temperature" refers to hardened oil having a melting
point of about 20° C. or more. The melting point may be 30° C. or more,
40° C. or more, or 50° C. or more. Hardened oil having a melting point of
20 to 50° C. is preferably used. What kind of the hardened oil to be used
can be determined in view of the storage temperature, the sowing time or
As the hardened oil, triglycerides, diglycerides or the like that contains
fatty acids of medium carbon chain can be used. Specific examples thereof
include, but are not limited to, butter, margarine, shortening, and cocoa
It is necessary that this inner membrane is waterproof, not only to form a
capsule enclosing an aqueous substance, but also to prevent from
evaporating water of the innermost layer (aqueous phase) that contains
"Biodegradability" or "microbiologically degradable" means that the
substance can be degraded or assimilated by microorganisms or other
organisms when being sown in soil. When the hardened oil is degraded or
assimilated, the inner membrane composed of the hardened oil is
decomposed. Thus the barrier against a release of the tissue to outside of
the capsule is removed, and the permeability of oxygen is increased, which
activates the cell tissue to get out from the dormant state so that
differentiation and growth of the tissue may occur.
It is preferable that the outer membrane is swollen with water and is
biodegradable or microbiologically degradable, because a barrier against
germination can be removed so that the cell tissues can be coming out form
the capsule after the differentiation and growth of the cell tissue.
Examples of the outer membranes that are biodegradable or
microbiologically degradable include proteins, polysaccharides, and
biodegradable plastics. These substances can be used alone or in
combination of two or more.
Examples of the proteins include, but are not limited to, gelatin and
collagen. These substances can be used alone or in combination of two or
As the polysaccharides, gel-forming polysaccharides are preferably used.
Examples of such polysaccharides include, but are not limited to, agar,
carrageenan, Arabian gum, gellant gum, xanthan gum, pectin, and alginic
acid. These substances can be used alone or in combination of two or more.
Examples of biodegradable plastics include, but are not limited to,
polylactic acid, polyhydroxybutyric acid, and a mixture thereof. These
substances can be used alone or in combination of two or more.
Furthermore, if desired, properties of the outer membrane can be modified
by adding saccharides, sugar alcohol, polyhydric alcohol, pullulan,
chitosan or the like to the above-described biodegradable substances.
A particularly preferable outer membrane is gelatin, whose oxygen barrier
properties can be increased after it is dried.
The artificial seed capsules of the present invention can have a structure
of four or more layers, if desired. By selecting materials for the outer
membrane, various properties can be given to the outer membrane.
Furthermore, the surface of the artificial seed capsules may be coated
A general method for producing a three-layered seamless soft capsule has
already been described above food application. For artificial seeds, for
example, as the inclusion (the innermost layer) of the capsule, a suitable
size of adventitious embryo which is obtained from a cell tissue (e.g.,
adventitious embryo) that is grown in a liquid medium or a solid medium
and suspended in a medium can be used. As the inner membrane, hardened oil
that is solid at room temperature can be used. As the outer membrane, a
solution of biodegradable substance having an appropriate concentration
(e.g., 22% gelatin solution) can be used. Using those components, the
three-layered capsule can be produced by dropping a drop of capsule into a
liquid using an apparatus provided with a triple tube nozzle. In this
case, a pump is adjusted so that one adventitious embryo is put in the
inner liquid in each capsule. Shaping and solidification of the capsule
are performed in solidification oil, so that a capsule having a
three-layered structure is produced. The solidification oil can be removed
from the capsules formed, and then the capsules are subjected to drum
drying, so that artificial seeds of the surface-dried capsule including
adventitious embryos are obtained. The drying is performed at a low
temperature so that the cell tissues do not die.
The particle size of the artificial seed capsule is 1 mm to 12 mm,
depending on the size of the cell tissues. Preferably, the size is 3 to 10
mm. The number of the cell tissues to be enclosed in a capsule can range
from one to several, depending on the size of the tissue. The number of
the tissues can be determined so that the capsule has appropriate
properties as the artificial seeds in view of the redifferentiation rate
(germination rate). The number of cell tissues depends on the type of the
plant but is preferably 1 to 4.
The thus obtained artificial seed capsules of the present invention can be
stored for three months or more at room temperature in a dry barn or the
like while retaining the germination ability. Therefore, there is no need
of a special refrigerator or storage in low temperature water. If they are
stored in a refrigerator at 10° C. or less, long term storage of 6 months
or more can be achieved.
The capsulated artificial seed of the present invention can be a hollow
seamless soft capsule encompassing a redifferentiable plant cell tissue
(cell tissue). The capsule is composed of an inner membrane made of
hardened oil and an outer membrane made of biodegradable gel (protein,
polysaccharides) or biodegradable plastics, where the surface of the
capsule (outer membrane) is dried. Such a composition of the capsule
restricts oxygen supply to the inside of the capsule, which enables the
cell tissue to survive in a dormant state in the capsule. Further, water
is prevented from evaporating from the capsule and the capsule can be
stored for two months or more at room temperature. Once the capsule is
sown in soil, not only the outer membrane but also the hardened oil of the
inner membrane can be degraded by water and the activities of the
microorganisms in soil, thereby the permeability of oxygen can be
increased and the cell tissue can be awakened from the dormant state and
transfers to the germination state.
Claim 1 of 3 Claims
1. A seamless capsule in which
living cell or tissue is suspended in an aqueous liquid,
wherein the capsule is formed by a seamless capsule manufacturing apparatus
with a triple tube nozzle,
wherein the aqueous liquid comprising the living cell or tissue is supplied
in an innermost tube of the triple tube nozzle,
wherein a lipophilic material is supplied in a middle tube of the triple
wherein an outer membrane forming material is supplied in an outermost tube
of the triple tube nozzle,
wherein the lipophilic material is selected from the group consisting of
deep frying oils, salad oil, vitamin E, wheat germ oil, sesame oil, hardened
oil having a melting point of 35° C. or less, cocao butter, butter,
margarine, shortening, and fatty acid esters of sucrose,
wherein the outer membrane forming material is at least one natural polymer
selected from the group consisting of gelatin, agar, pectin, alginic acid,
alginate, carrageenan, curdlan, starch, gellan gum, glucomannan, and
mixtures thereof, and
wherein the cell or the tissue can grow in the aqueous liquid.
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