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, polysaccharides, etc.

This application is the National Stage if International Application No. PCT/JP01/05606, filed Jun. 28, 2001.

TECHNICAL FIELD

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 in soil.

BACKGROUND ART

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 intestine.

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 technique.

DISCLOSURE OF INVENTION

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 the liquid.

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 outer membrane.

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 present invention.

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 spherical shape.

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 tissues.

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 capsule.

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 or pectin.

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 microorganisms.

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 the like.

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 butter.

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 cell tissues.

"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 more.

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 with pharmaceuticals.

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 tube nozzle,

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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