JP2009136201A - Method for preserving biological component - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for preserving a biological component immobilized without freezing in a medium consisting essentially of water under mild conditions to a biological component of normal temperature and normal pressure. <P>SOLUTION: The method for preserving a biological component comprises mixing a water-soluble polymer solution into which a biological component is dispersed with a water-soluble compound solution to form a three-dimensional crosslinked product and preserving the biological component in the state of the three-dimensional crosslinked product. Preferably the water-soluble polymer is a polymer containing a phosphorylcholine group and a phenylboronic acid group and the water-soluble compound is a compound containing a polyfunctional hydroxy group. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a method for holding and storing biological components and a method for transporting.

Generally, immobilization of living components such as cells and tissues in an alive state is an important technique for the industrialization of biotechnology and regenerative medicine based on biotechnology and bioengineering. In general, animal cells, which are one of the biological components, are generally stored in a cryopreservation solution by dispersing the cells and tissues in a cryopreservation tube made of glass or plastic. Patent Document 1).
Specifically, cell culture solutions and serums prepared by the workers themselves and those containing 10% dimethyl sulfoxide and methyl cellulose added thereto, and preservation solutions with specially prepared formulations are commercially available.
In addition, a technique for storing cells while retaining them on a plate has been developed (see Patent Document 1).

In general, cells are dispersed in these storage solutions and sealed in liquid nitrogen and then stored in liquid nitrogen. However, when freezing, they should not be frozen quickly, but gradually frozen at lower temperatures. There is a need. For this purpose, a dedicated freezing apparatus is commercially available that gradually supplies liquid nitrogen and lowers the temperature by 1 ° C. from 1 ° C. per minute to 2 ° C.
Since such an apparatus is expensive, the temperature drop when isopropyl alcohol is put in an ultra-low temperature freezer below -80 ° C. is close to 1 ° C. for 1 minute. There is a method of freezing in a refrigerator (4 ° C.), then in a freezer at −20 ° C. for 1 hour, and in an ultra-low temperature freezer at −80 ° C. overnight.
Thus, even if the temperature is strictly controlled and stored in liquid nitrogen, there are quite a few cells with a very poor cell viability after thawing (less than several percent, almost no cells survive). In particular, there are many cells that have important functions such as primary cells, germ cells, fusion cells, and transgenic cells. Since these cells cannot be preserved, the necessary experiments cannot be performed when necessary.
In addition, it is known that the cell activity deteriorates over time unless cells after thawing are immediately seeded in a culture vessel and the culture is started.

Dimethyl sulfoxide itself is toxic to cells and is not preferable to be exposed for a long time, and some cells may be denatured by dimethyl sulfoxide added during freezing (human-derived acute myeloid leukemia) The cell line HL-60 cell is induced to differentiate into neutrophil-like cells by dimethyl sulfoxide), and is a reagent that should not be used originally. Therefore, various cell cryopreservation solutions have been researched, developed, and sold. Yes.
Many efforts have been made to preserve cells and tissues through such efforts, but there are many cells that cannot be cryopreserved. Stable culturing of these cells contributes to the development of biotechnology. .
In addition, in order to stably store biological components, it is necessary to prevent denaturation that frequently occurs during storage.

Normally, cells cannot be stored at 4 to 37 ° C., and even when the above cell storage solution is added, cell death occurs from the beginning of storage, and most cells die within 24 hours. At 37 ° C., cells can only be maintained by culturing, but culturing requires daily observation, medium exchange, etc., and cannot be said to be preserved. In addition, it is difficult to preserve the cells because the cells are altered during the culture, or there are cells that cannot be cultured for a long time.
The only way to maintain biological components such as cells and tissues is by cryopreservation at ultra-low temperatures, but there are many biological components that cannot be stored, and there is a craving for a method that can stably store and retain biological components. Has been. For example, in the cells, cell groups that have undergone functional differentiation such as liver parenchymal cells, pancreatic islet cells, microglia cells, stem cell groups such as hematopoietic stem cells, human ES cells, human iPS cells, monkey ES cells, eggs, EC cells, etc. It is difficult to save. These cells are very useful cells, and development of a stable storage method is desired.

“Cultured Cell Experiment Handbook”, Yodosha, 2004, P69-74 JP 2001-16626 A

  The object of the present invention is to use conditions that are mild to biological components such as normal temperature and normal pressure without using physical methods such as chemical reaction, heat, light, and radiation irradiation in a medium mainly composed of water. The object is to provide a method for preserving an immobilized biological component without freezing.

The present invention is as follows.
(1) A water-soluble polymer solution in which a biological component is dispersed and a water-soluble compound solution are mixed to form a three-dimensional crosslinked body, and stored in the state of the three-dimensional crosslinked body. Preservation method.
(2) The method for preserving a biological component according to (1), wherein the water-soluble polymer is a polymer containing a phosphorylcholine group and a phenylboronic acid group.
(3) The method for preserving a biological component according to (1) or (2), wherein the water-soluble compound is a compound having a polyvalent hydroxyl group.
(4) Any one of (1) to (3), wherein the biological component is at least one selected from a cell, a cell three-dimensional tissue constructed by the cell, a germ cell, a genetically modified cell, and a biological tissue. Method of preserving the biological components.
(5) A water-soluble polymer solution in which a biological component is dispersed and a water-soluble compound solution are mixed to form a three-dimensional crosslinked body, and the biological component is transported in the state of the three-dimensional crosslinked body Transport method.
(6) The method for transporting a biological component according to (5), wherein the water-soluble polymer is a polymer containing a phosphorylcholine group and a phenylboronic acid group.
(7) The method for transporting biological components according to (5) or (6), wherein the water-soluble compound is a compound having a polyvalent hydroxyl group.
(8) Any one of (5) to (7), wherein the biological component is at least one selected from a cell, a cell three-dimensional tissue constructed by the cell, a germ cell, a genetically modified cell, and a biological tissue. For transporting biological components.
(9) The biological component is recovered by releasing the crosslinked structure by adding a compound having a hydroxyl group to the three-dimensional crosslinked body stored by the biological component storage method according to any one of (1) to (4). A method for recovering biological components.
(10) The biological component is recovered by adding a compound having a hydroxyl group to the three-dimensional crosslinked body transported by the biological component transport method according to any one of (5) to (9) to release the crosslinked structure. A method for recovering biological components.

  By storing the biological component according to the present invention, unlike the conventional storage method by freezing, damage to the biological component due to freezing and thawing can be reduced, and the biological component that could not be cryopreserved can be stored stably. It becomes possible. As a result, the biological components immobilized at around normal temperature without freezing can maintain high activity and can be effectively used for storage and transportation of cells, tissues and the like in the bio industry and regenerative medicine.

  The present invention is a method for preserving and transporting a biological component that is formed by mixing a water-soluble polymer solution in which a biological component is dispersed and a water-soluble compound solution to form a three-dimensional crosslinked body and storing and transporting the three-dimensional crosslinked body in the state. It is a transportation method.

  The water-soluble polymer used in the present invention is preferably a polymer containing a phosphorylcholine group and a phenylboronic acid group.

  A polymer containing a phosphorylcholine group and a phenylboronic acid group at the same time is produced by mixing a monomer containing a phosphorylcholine group and a monomer having a phenylboronic acid group in a liquid state and performing a radical polymerization reaction in the presence of a radical generator. be able to. Further, for the purpose of imparting the polymer with binding properties to the container base material when gelled, it is possible to add other monomers if necessary to form a multi-component copolymer.

The monomer containing a phosphorylcholine group is preferably a compound having a carbon-carbon double bond such as a vinyl group or an allyl group as a polymerizable group and having a phosphorylcholine group in the same molecule.
For example, 2-methacryloyloxyethyl phosphorylcholine, (2-methacryloyloxylethyl-2 ′-(trimerityl ammonia) ethyl phosphate, 2- (meth) acryloyloxyethyl-2 ′-(trimethylammonio) ethyl phosphate, and the like can be given. However, 2-methacryloyloxyethyl phosphorylcholine (MPC) is particularly preferable.

The monomer having a phenylboronic acid group is preferably a compound having a carbon-carbon double bond such as a vinyl group or an allyl group as a polymerizable group and having a phophenylboronic acid group in the same molecule.
For example, p-vinylphenylboronic acid, m-vinylphenylboronic acid, p- (meth) acryloyloxylphenylboronic acid, m- (meth) acryloyloxyphenylboronic acid, p- (meth) acrylamidophenylboronic acid, p- Examples thereof include vinyloxyphenyl boronic acid, m-vinyloxyphenyl boronic acid, vinyl urethane phenyl boronic acid, and p-vinyl phenyl boronic acid or m-vinyl phenyl boronic acid is preferable.

  Examples of other monomers include (meth) acrylic acid, sodium (meth) acrylate, 2-hydroxyethyl (meth) acrylate, glycerol (meth) acrylate, N-vinylpyrrolidone, acrylonitrile, (meth) acrylamide, and vinylbenzene. Hydrophilic monomers such as sodium sulfonate, hydrophobic monomers such as methyl (meth) acrylate, ethyl (meth) acrylate, butyl (meth) acrylate, lauryl (meth) acrylate, dodecyl (meth) acrylate, styrene, vinyl acetate, ( 3-methacryloyloxypropyl) monomers having an alkyloxysilane group such as trimethoxysilane, monomers having a siloxane group, monomers having a glycidyl group such as glycidyl methacrylate, allylamine, aminoethyl (Meth) A monomer having an amino group such as acrylate, a carboxyl group, a hydroxyl group, an aldehyde group, a thiol group, a halogen group, a methoxy group, an epoxy group, succinimide group, and a monomer having a maleimide group.

  Among these, a particularly preferable monomer combination is a terpolymer of 2-methacryloyloxyethyl phosphorylcholine (hereinafter abbreviated as MPC), p-vinylphenylboronic acid, and butyl (meth) acrylate.

  As the radical generator, aliphatic azo compounds such as azobisisobutyronitrile, 4,4′-azobis (4-cyanopentanoic acid), and peroxides such as benzoyl peroxide and succinic acid peroxide are preferable.

  The composition mole fraction in the polymer can be controlled by the composition of the monomer mixed solution. The range of the molar fraction of the monomer unit having a phosphorylcholine group is 0.01 to 0.99, preferably 0.30 to 0.70, more preferably 0.45 to 0.60. is there. The range of the molar fraction of the monomer unit having a phenylboron group in the polymer is 0.01 to 0.99, preferably 0.03 to 0.50, and more preferably 0.30 to 0.00. A range of 40.

  The molecular weight of the polymer is measured by gel permeation chromatography, and polyethylene oxide is converted as a standard substance. The range is 1,000 to 10,000,000, preferably 10,000 to 1,000,000. 000, more preferably 20,000 to 300,000. When the molecular weight exceeds the upper limit value, the solubility in an aqueous medium may be reduced.

  The water-soluble compound used in the present invention is preferably a compound having a polyvalent hydroxyl group. The compound having a polyvalent hydroxyl group needs to be dissolved in an aqueous solvent to form a uniform solution. Examples include monosaccharides, disaccharides, polysaccharides, low molecular weight polyhydric alcohols, polyvinyl alcohol, etc., but selecting polysaccharides or polyvinyl alcohol (PVA) constitutes a three-dimensional cross-linked body with a stable structure. This is preferable.

A three-dimensional crosslinked product can be produced by mixing an aqueous solution containing a polymer containing a phosphorylcholine group and a phenylboronic acid group simultaneously with an aqueous solution containing a polyvalent hydroxyl compound.
In order to capture and stably store biological components in the three-dimensional crosslinked body, the solvent should be aqueous and the use of organic solvents should be avoided as much as possible. Moreover, it is desirable that the pH of the solvent that does not cause a physiological problem is in the range of 7.0 to 8.0.

  The polymer containing a phosphorylcholine group and a phenylboronic acid group and a polyhydric hydroxyl group compound can be used in the range of being soluble in an aqueous solvent, but can be used in the range of 0.5 to 20% by weight, preferably 1. A range of 5 to 10% by weight is preferred.

  The temperature for forming the three-dimensional crosslinked body is preferably 0 to 37 ° C. and more preferably 4 to 37 ° C. in order to preserve biological components. If the temperature is outside this range, the biological components may be damaged.

Biocomponents can be encapsulated in a three-dimensional crosslinked body by dispersing a biological component in a water-soluble polymer solution in advance and mixing an aqueous solution containing a polyvalent hydroxyl compound in the dispersion to form a three-dimensional crosslinked body. it can.
Conversely, a biological component can be dispersed in advance in an aqueous solution containing a polyvalent hydroxyl compound, and a water-soluble polymer can be mixed with the dispersion to form a three-dimensional crosslinked product. In addition, a three-dimensional crosslinked body can be constructed by mixing a water-soluble polymer solution and an aqueous solution containing a polyvalent hydroxyl compound on cells cultured in a culture vessel, and the cells can be embedded in the three-dimensional crosslinked body. .

  Examples of biological components used in the present invention include cells, cell three-dimensional tissues constructed by cells, germ cells, genetically modified cells, tissues derived from living bodies, and the like.

  In the present invention, the temperature for storing or transporting the biological component encapsulated in the three-dimensional crosslinked body is preferably 0 to 37 ° C, and more preferably 4 to 37 ° C.

  By adding a compound having a hydroxyl group to the three-dimensional crosslinked product stored and transported by the method for storing and transporting biological components of the present invention, it is possible to recover the biological components by releasing the crosslinked structure. Glucose etc. are mentioned as a compound which has a hydroxyl group.

(Synthesis of polymer containing phosphorylcholine group and phenylboronic acid group simultaneously)
In a flask, 5.3 g of 2-methacryloyloxyethyl phosphorylcholine (MPC) was weighed, charged with 30 g of ethanol, and the inside of the container was purged with argon gas while stirring. 0.44 g of p-vinylphenylboronic acid (VPBA), 1.3 g of n-butyl methacrylate (BMA) and 0.05 g of 2,2′-azobisisobutyronitrile were added and stirred uniformly. After sealing, the mixture was heated to 60 ° C. and reacted for 48 hours. The obtained reaction solution was added dropwise to a diethyl ether / chloroform (8/2) solution to obtain a solid polymer. As a result of NMR analysis of the obtained polymer, it was MPC / VPBA / BMA = 53/11/36 (molar ratio). The number average molecular weight was 27,000. This polymer is designated as PMBV.
(Polyhydric hydroxyl compound)
A commercially available polyvinyl alcohol (PVA) having an average polymerization degree of 1500 (manufactured by Wako Pure Chemicals, 160-03055) was used.

The following examples and comparative examples were carried out by aseptic work in a clean bench unless otherwise stated because of experiments for handling cells.
Example 1
PVA was dissolved in heated ultrapure water to prepare a 5 wt% solution. This was sterilized by filtration through a 0.2 μm sterilized filtration filter (Toyobo Co., Ltd., ADVANTEC3912212). PMBV was dissolved in Dulbecco's modified Eagle's medium (DMEM, manufactured by Invitrogen, 11885-084) containing 10% fetal bovine serum (Invitrogen, 10099-141) and sterilized by filtration.
The cells used were human liver cancer-derived cells, HepG2 (obtained from DS Pharma Biomedical, EC85011430). HepG2 cells were cultured in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum in a cell culture flask (Sumitomo Bakelite, MS-21250). When the cells became semi-confluent, the cell surface was washed with a sterilized phosphate buffer, D-PBS (Invitrogen, 14190-144), and 0.25% trypsin / EDTA 1 mL (Invitrogen, 25399-054). And reacted at 37 ° C. for 5 minutes. After collecting the cells, the number of viable cells was counted with a hemocytometer. Cells were dispersed in a PMBV solution to prepare 1 × 10 ⁵ cells / 1 mL.
0.25 mL of 5 wt% PVA solution was placed in each well of a 24-well plate for cell culture (manufactured by Sumitomo Bakelite, MS-80240), and the solution was made uniform on the bottom of the well. An equal amount of the PMBV solution in which the cells were dispersed was added from above, and the plate was immediately tilted to mix the PVA solution and the PMBV solution, which was then placed in a 37 ° C. 5% carbon dioxide incubator. Three-dimensional crosslinking (gelation) occurred immediately after mixing and was completely gelled by standing for 30 minutes. It was confirmed by observation with an optical microscope that HepG2 cells were retained in the three-dimensional crosslinked body (gel).
This gel was stored at 37 ° C. for 4 weeks.
The photograph after storage for 4 weeks is shown on the left of FIG. In the gel, the cells existed in a spherical shape, and when the gel was embedded, the cells were dispersed in a single cell, but a certain amount of colonies were formed. However, as shown in the right of FIG. 1, the HepG2 cells seeded on a 35 mm diameter petri dish on the same day and stored for 4 weeks were over-proliferated and already killed, indicating that they were stored in the gel. .

Example 2
The gel under storage in Example 1 was dissolved by the following method, and the cells were collected and adhesion culture was performed.
Commercially available 40% D-glucose (Sigma Aldrich, G8769) was prepared to 10% with D-MEM. The gel during storage was cut with an autoclave-sterilized surgical scissors. The previously prepared 10% D-glucose solution was added to the gel fragments to dissolve the gel.
The gel lysate was placed in a centrifuge tube and centrifuged at 1000 rpm for 1 minute, and the cells were collected as a pellet. D-MEM was added to the obtained cell pellet to prepare a new cell dispersion. After the number of cells was adjusted to 1 × 10 ⁵ cells / mL, the cells were seeded on a 35 mm diameter tissue culture dish and re-culture was started.
FIG. 2 shows the observation results of seeding and recovering the cells recovered by dissolving the gel after storage for 2 weeks (left is culture day 1 and right is culture day 3). It was confirmed not only that the cells were alive and adhered and extended, but also that cell proliferation occurred from the observation image on the third day, and that the cell function was preserved.

Example 3
The same PVA solution as in Example 1 was used. PMBV was dissolved in vascular endothelial cell growth medium, HuMedia-EG2 (manufactured by Kurabo Industries, KE-2150S) to a concentration of 5% by weight and sterilized by filtration.
Human vascular endothelial cells used were cultured by culturing commercially available human umbilical vein-derived vascular endothelium (Kurabo, KE-4109) in a cell culture flask (Sumitomo Bakelite, MS-21250) using HuMedia-EG2. Cells were collected in the same manner as in Example 1. After counting the number of cells, it was adjusted to 5 × 10 ⁴ cells / mL with a PMBV solution.
0.25 mL of a 5% PVA solution is placed in each well of a 24-well plate in advance, and further, 0.25 mL of the PMBV solution in which cells are dispersed is added on each well and mixed, and then in a 37 ° C. 5% CO 2 incubator. Gelled.
After gelation, it was stored at 37 ° C. in the same incubator. The medium was not changed.
Two weeks later, the gel was dissolved according to the method of Example 2, cells were collected, and the number of cells and the survival rate were measured. The collected cells were cultured by a conventional method, and the cell behavior was examined.
As shown on the left of FIG. 3, the human vascular endothelial cells after being stored for 2 weeks maintained a spherical shape. In addition, as shown on the right side of FIG. 3, it was confirmed that the cells recovered by dissolving the gel after storage for 2 weeks could be continuously cultured after the gel storage with almost no change from the normal form.

<< Comparative Example 1 >>
Gel-embedded culture using a collagen type I 0.3% solution (Koken, 1-PC) was performed.
Collagen was gelled by using the reconstitution function of collagen fibers in the neutral pH region, and the cells were dispersed to form a gel. Specifically, in a sterilized container, 8 parts of collagen type I solution was cooled with ice, and 1 part of F12 medium (Invitrogen, 11765-054) prepared to a 10-fold concentration, 1 part of sodium hydroxide HEPES solution Were collected under ice age, and the collected human vascular endothelial cells collected in Example 3 were dispersed therein and seeded in a 24-well plate. Subsequently, it was gelatinized by heating at 37 ° C. for 30 minutes. As it was, it was then cultured at 37 ° C. in an incubator. The medium was changed every other day.
Two weeks later, Hank's balanced salt solution (manufactured by Invitrogen, 24020-117) containing 0.5% collagenase (manufactured by Wako Pure Chemical Industries, Ltd., 031-17601) was added to the collagen gel and heated at 37 ° C. to dissolve the gel. After lysis, the lysate was transferred to a centrifuge tube and centrifuged at 1000 rpm for 1 minute to collect cells as pellets. The cell pellet was washed with DMEM containing 10% fetal bovine serum, dispersed again in DMEM, and the number of viable cells was counted.

  As shown in Table 1, the cell survival rate when retained in the three-dimensional crosslinked body was 89%, and a significant difference was observed with the cell survival rate of 55% when stored in a collagen gel. This indicates that cell damage was caused by using an enzyme in Comparative Example 1 when the gel was dissolved, and the superiority of storage by the three-dimensional crosslinked product according to the present invention was shown.

Example 4
First, HepG2 cells were cultured on a 90-mm diameter petri dish using D-MEM containing 10% fetal bovine serum to proliferate the cells. Next, proliferated HepG2 cells were collected using trypsin / EDTA. Count the number of cells with a hemocytometer and dispense 2 mL of the cell dispersion prepared to 10,000 mL / medium 1 mL into a Sumilon Celtite X (Sumitomo Bakelite, MS-9035X) 35 mm diameter petri dish, The cells were cultured in an incubator at 37 ° C. and 5% carbon dioxide for 3 days to form spheroids formed by cell aggregation.
The same PVA solution as in Example 1 was used. PMBV was dissolved in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum to 5% by weight and sterilized by filtration.
Spheroids were collected, centrifuged, and prepared into a spheroid dispersion with a PMBV solution.
0.25 mL of a 5% PVA solution is put in advance in each well of a 24-well plate, and further, 0.25 mL of a PMBV solution in which spheroids are dispersed is added and mixed, and then a 37 ° C., 5% carbon dioxide incubator. Gelled inside.
After gelation, it was stored at 37 ° C. in the same incubator. The gel was dissolved according to the method of Example 2 to collect spheroids, and the number of cells and the survival rate were measured.

<< Comparative Example 2 >>
Gel embedding culture using a collagen type I 0.3% solution (manufactured by Koken, 1-PC) was carried out as a comparative example.
A collagen gel was prepared using spheroids as in Comparative Example 1.
Two weeks later, Hank's buffer solution (manufactured by Invitrogen) containing 0.5% collagenase (manufactured by Wako Pure Chemical Industries, Ltd.) was added to the collagen gel and heated at 37 ° C. to dissolve the gel. After dissolution, the lysate was transferred to a centrifuge tube and centrifuged at 1000 rpm for 1 minute to collect spheroids. The spheroids were washed with DMEM containing 10% fetal bovine serum, dispersed again in DMEM, and the number of viable cells was counted.

  As shown in Table 2, the cell survival rate when retained in a three-dimensional crosslinked body was 92%, showing a significant difference from the cell survival rate of 65% when stored in a collagen gel. This indicates that in Comparative Example 2 cell damage was caused by using an enzyme when the gel was dissolved, and the superiority of spheroid storage by the three-dimensional crosslinked product according to the present invention was shown.

  According to the present invention, it is possible to immobilize biological components in a three-dimensional crosslinked body at around room temperature without freezing, the immobilized biological components can maintain high activity, cells in the bio industry and regenerative medicine, It can be used effectively for preservation of organizations.

Photographs of cell morphology of three-dimensional cross-linked bodies of HepG2 cells (preservation in gel) and normal culture (35 mm diameter petri dish culture) Photograph of re-cultured form of HepG cells recovered from 3D cross-linked body Photograph of re-cultured form of human vascular endothelial cells in 3D cross-linked body and cells recovered from 3D cross-linked body

Claims (10)

A method for preserving a biological component, comprising mixing a water-soluble polymer solution in which a biological component is dispersed and a water-soluble compound solution to form a three-dimensional crosslinked body, and storing the three-dimensional crosslinked body. The method for preserving a biological component according to claim 1, wherein the water-soluble polymer is a polymer containing a phosphorylcholine group and a phenylboronic acid group. The method for preserving a biological component according to claim 1 or 2, wherein the water-soluble compound is a compound having a polyvalent hydroxyl group. The biological component preservation according to any one of claims 1 to 3, wherein the biological component is at least one selected from a cell, a cell three-dimensional tissue constructed by the cell, a germ cell, a genetically modified cell, and a biological tissue. Method. A method for transporting a biological component, comprising mixing a water-soluble polymer solution in which a biological component is dispersed and a water-soluble compound solution to form a three-dimensional crosslinked body, and transporting the three-dimensional crosslinked body. The method for transporting biological components according to claim 5, wherein the water-soluble polymer is a polymer containing a phosphorylcholine group and a phenylboronic acid group. The method for transporting biological components according to claim 5 or 6, wherein the water-soluble compound is a compound having a polyvalent hydroxyl group. The transport of the biological component according to any one of claims 5 to 7, wherein the biological component is at least one selected from a cell, a cell three-dimensional tissue constructed by the cell, a germ cell, a genetically modified cell, and a tissue derived from a living body. Method. A biological component, wherein a biological component is recovered by releasing a crosslinked structure by adding a compound having a hydroxyl group to the three-dimensional crosslinked body stored by the biological component storage method according to any one of claims 1 to 4. Recovery method. A biological component, wherein a biological component is recovered by adding a compound having a hydroxyl group to the three-dimensional crosslinked body transported by the biological component transport method according to any one of claims 5 to 9 to release the crosslinked structure. Recovery method.

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