JPS61276858A - Biomedical material - Google Patents

Abstract

PURPOSE:A biomedical material, having amphoteric ion groups, consisting of a high polymer having a specific surface zeta potential, having biocompatibility and used for cultivating, separating and purifying cells as a cell adsorbent. CONSTITUTION:A biomedical material obtained by polymerizing an amphoteric ionic compound, e.g. lauryldimethylbetaine, with an ethylenically unsaturated monomer, e.g. methyl (meth)acrylate, at 1:99-50:50, or blending or chemically bonding the amphoteric ionic compound to a substrate high polymer at 1:99-50:50 ratio and consisting of a high polymer material having -75-+75mV, preferably -60-+60mV surface zeta potential. The above-mentioned high polymer material is used as particles having 0.01-1,000mu average particle diameter or film for cultivating, separating or purifying cells.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel biomedical material and a method for treating cells using the material as a cell adsorbent.

′ and − In recent years, techniques for genetic manipulation, cell fusion, and mass cell culture have been attracting attention for the purpose of improving biological functions and cultivars. However, these success stories are rare. This is because each technology has many problems. Current challenges in mass culture of animal and plant cells include the fact that animal and plant cells multiply much more slowly than microbial cells, especially bacteria, and are more susceptible to changes in the environment. It is also necessary to separate large quantities of cells obtained using techniques such as genetic manipulation and cell fusion.

Cells and blasts, which are cells with cell walls removed, are vulnerable to environmental changes (e.g. temperature, pH, pressure, etc.) in an isolated state, but are significantly stabilized when adsorbed to other objects. have.

Therefore, if treatments such as culturing, separation, and purification are performed in an adsorbed state, the treatment becomes significantly easier. However, no suitable adsorbent is known so far that allows cells to be treated in an adsorbed state.

The present invention provides a biocompatible biomedical material comprising a polymeric material having a zwitterionic group and having a surface zeta potential of -6° to +60 mV, preferably -6° to +60 mV. .

Preferably the form of the biomedical material has an average particle size of 0.01
The resin particles of ~1000 μm are also membranes (including membranes formed on the inner walls of containers such as flasks and petri dishes, and membranes covering chromatographic carriers such as silica gel).

The polymer material containing the zwitterionic group is obtained by polymerizing an ethylenically unsaturated monomer in the presence of a zwitterionic compound)
, or by chemically bonding or blending a zwitterionic compound to a substrate polymeric compound.

The present invention also provides a treatment method for cell culture, isolation, purification, etc., using the biomedical material as a cell adsorbent.

Cells and protoplasts, which are obtained by removing the cell wall, usually have a surface charge (expressed as zeta potential), and the surface is mainly composed of unevenly distributed proteins, phosphate groups derived from glycoproteins, It is thought that the structure has prominent carboxyl groups, amino groups, and sulfate groups. The relationship between adsorption between cells having such a surface structure and the surface properties of membranes or particles is still largely unknown. The present invention revealed for the first time that adsorption between membranes or particles and cells is controlled by zwitterionic groups and surface Zeta potential. The feature of the present invention is that the zwitterionic group and the −75 to +75
The object of the present invention is that cells can be adsorbed to a polymer compound having a surface zeta potential of mV to facilitate cell injection, cell culture, and cell separation.

■Biomedical materials in the present invention refer to living organisms and materials isolated from living organisms, such as biological cells, biological tissues, cell granules, enzymes, and substances that come into contact with physiologically active substances, and especially materials that come into contact with cells. This is particularly desirable when applying the present invention.

Depending on the degree of contact with cells, biomedical materials can be classified into cell culture materials, cell adsorption materials, cell separation materials, etc.

The cells may be microbial cells, animal cells, plant cells, or hybrid cells of these cells. It also includes protoplasts obtained by removing the cell walls of these cells. Protoplasts include fused protoplasts, which are formed by fusion of two or more protoplasts, and modified protoplasts, which are formed by injecting genetic materials into protoplasts.

It has a zwitterionic group and has a surface zeta potential of 175 to +
Polymeric materials that are 75 mV can be produced by polymerizing ethylenically unsaturated monomers in the presence of zwitterionic compounds. Alternatively, the zwitterionic compound can be prepared by chemically bonding or blending the zwitterionic compound to a matrix polymeric compound.

The zwitterionic compounds used in the present invention include zwitterionic surfactants that have both anionic and cationic groups in the molecule, polymerizable zwitterionic monomers, oligomers and polymers that have zwitterionic groups. .

Examples of zwitterionic surfactants include, for example, "
Zwitterionic surfactants belonging to carboxylate type, sulfate ester salt type, sulfonate ester salt type, and phosphate ester salt type, described in "Introduction to New Surfactants", Sanyo Kasei Kogyo ■, pp. 81-89. agents can be used.

Among these, lauryl dimethyl betaine, stearyl dimethyl betaine, lauryl hydroxyethyl betaine, and the like are known to be commonly used.

In addition, 1-(3-sulfol 2), a product of RASHIG,
-hydroxypropyl)pyridinium betaine, 1-(
3-sulfopropyl)pyridinium betaine, lauryldimethylammonium 3-sulfopropylbetaine, myristyldimethylammonium 3-sulfopropylbetaine, palmityldimethylammonium 3-sulfopropylbetaine, stearyldimethylammonium 3-sulfopropylbetaine.

Alkylamidopropyldimethylammonium 3-sulfoprobyl betaine and the like can also be used.

Furthermore, N-(2-hydroxydodecyl)-2-aminoethanesulfonic acid, which is a product of Nippon Paint ■.

N-(2-hydroxytriacontyl)-2-aminoethanesulfonic acid, N-methyl-N-(2-hydroxydodecyl)-2-aminoethanesulfonic acid, N-(2-
Hydroxy-3-(α-oxydodecyloxy)propyl)-2-aminoethanesulfonic acid, lauryl aminopropionic acid, etc. can also be used.

Examples of polymerizable zwitterionic finishing agents include compounds of the general formula type, such as N-(3-sulfopropyl)-N-methacryloxyethyl-N,N-dimethylammonium betaine, and compounds of the general formula type, For example 1-(3-sulfopropyl)
-2-vinylpyridinium betaine, etc., and the formula % formula % (wherein R1 is a group or ^, and R, R'R" and R'" are hydrogen or an alkyl group, R2 is H, Cl-211 alkyl group, and the alkyl group has 15o-, -coo-,
may contain -o-, R3 may be substituted with an alkyl group (J-12 is an alkylene group or a phenylene group, and A represents -COOH or -3OaH). Amino acid compounds, such as N-methyl-
N-(vinylbenzyl)taurine and the like can be used.

As an oligomer having a zwitterionic group, formula % (wherein R is a 01-6 alkylene or phenylene group which may have a substituent, A is -COOH or -5O3
Resin oligomers having amphoteric ionic groups represented by H group), oil-free polyesters containing amphoteric ionic groups disclosed in JP-A-57-21927 (Polymer Emulsifiers), JP-A-57-40522 ( Examples include modified epoxy resins disclosed in "Water Dispersed Modified Epoxy Resins" and acrylic oligomers obtained by polymerizing ethylenically unsaturated monomer mixtures containing polymerizable amino acid compounds.

As the polymer having an amphoteric ionic group, oil-free polyester resins of the same type as the oligomers, modified epoxy resins, acrylic resins, etc. can be used.

Particles of a polymeric material having a zwitterionic group can be obtained by copolymerizing an ethylenically unsaturated monomer and the zwitterionic monomer by a method such as emulsion polymerization, precipitation polymerization, suspension polymerization, or bulk polymerization, or It can be produced by polymerizing the ethylenically unsaturated monomer according to the polymerization method described above in the presence of the zwitterionic surfactant or zwitterionic oligomer or polymer.

Examples of ethylenically unsaturated monomers include methyl (meth)acrylate, ethyl (meth)acrylate, n-butyl (meth)acrylate, isobutyl (meth)acrylate, and 2-ethylhexyl (meth)acrylate. Alkyl esters of acrylic acid or methacrylic acid and other monomers having ethylenically unsaturated bonds that can be copolymerized with them, such as styrene, α-methylstyrene, vinyltoluene, t-butylstyrene, ethylene, propylene, acetic acid Examples include vinyl, vinyl propionate, acrylonitrile, methacrylonitrile, and dimethylaminoethyl (meth)acrylate. Two or more types of these monomers may be used.

The ethylenically unsaturated monomer may contain a crosslinkable comonomer as part of its components in order to internally crosslink the resin particles.

The crosslinkable comonomer, which is an optional component, is a monomer having two or more radically polymerizable ethylenically unsaturated bonds in the molecule and/or two types each carrying a group that can react with each other. Contains ethylenically unsaturated group-containing monomers.

Monomers having two or more radically polymerizable ethylenically unsaturated groups in the molecule include polymerizable unsaturated monocarboxylic acid esters of polyhydric alcohols, polymerizable unsaturated alcohol esters of polybasic acids, and There are aromatic compounds substituted with one or more vinyl groups, examples of which include the following compounds.

Ethylene glycol diacrylate, ethylene glycol dimethacrylate, triethylene glycol dimethacrylate, tetraethylene glycol dimethacrylate, 1,3-butylene glycol dimethacrylate, trimethylolpropane triacrylate, trimethylolpropane tritaacrylate, 1,4-butane Diol diacrylate, neopentyl glycol diacrylate, 1,6-hexanediol diacrylate, pentaerythritol diacrylate, pentaerythritol triacrylate, pentaerythritol tetraacrylate, pentaerythritol dimethacrylate, pentaerythritol trimethacrylate, pentaerythritol tetramethacrylate, glycerol dimethacrylate, glycerol diacrylate, glycerol allyloxy dimethacrylate, 1,1.1-1-lishydroxymethylethane diacrylate, 1,1.1-)
Lishydroxymethylethane triacrylate, 1.1
．． 1-trishydroxymethylethane dimethacrylate, 1.1.1-1-lishydroxymethylethane trimethacrylate, 1,1.1-1-lishydroxymethylpropane diacrylate, 1,1.1-trishydroxymethylpropane trimethacrylate Acrylate, 1.1.1-trishydroxymethylpropane dimethacrylate, 1.1
．． 1-1-Lishydroxymethylpropane tritacrylate, triallyl cyanurate, triallyl isocyanurate, triallyl trimellitate, diallyl terephthalate, diallyl phthalate and divinylbenzene.

The ratio of the zwitterionic compound in the form of a monomer, surfactant, oligomer or polymer to the ethylenically unsaturated monomer may vary from 1:99 to 50:50.

A film of the zwitterionic polymer material can be formed by forming a film from a resin solution containing the zwitterionic polymer particles produced by the method described above, or from a solution of the zwitterionic polymer itself, or by forming a film from a solution of the zwitterionic polymer itself, or by forming a film from a resin solution containing the zwitterionic polymer particles produced by the method described above, or by forming a film from a solution of the zwitterionic polymer itself. It can be produced by forming a film from a resin obtained by chemically bonding to a polymer compound. For example, by using a zwitterionic compound having a hydroxyl group as part of the polyol component during the synthesis of polyester resin,
Further, amphoteric ionic oligomers and polymers having a crosslinking group such as a hydroxyl group can be chemically bonded to a substrate resin by reaction with a crosslinking agent such as a melamine resin or a polyisocyanate. Furthermore, if the zwitterionic compound is in a form such as an oligomer or polymer that cannot be eluted with water, it may be blended with a suitable base resin.

The ratio of zwitterionic compound to substrate polymeric compound can again vary from 1:99 to 50:50.

A membrane or other form of zwitterionic polymeric material may be ground to an appropriate particle size and classified for use as zwitterionic resin particles.

The polymer compound used in the present invention can be formed into a film or particles depending on the purpose of use.

For example, to form particles, an emulsion polymerization method is used when the average particle size is from 0.01 to 2μ, a precipitation polymerization method is used when the average particle size is from 0.2 to 10μ, and a suspension polymerization method is used when the average particle size is from 5 to 1000μ. In addition, in order to form a membrane, it is better to use a cell culture substrate, such as a culture device such as a Petri dish or flask, or on the surface of a microcarrier, which is more suitable for cell culture, cell adsorption, and cell separation. It becomes the base material. In addition, if it is carried out on the surface of a chromatographic carrier such as silica gel particles, the substrate becomes more suitable for cell separation. Particles with a small particle size are mainly used for cell culture, particles with a large particle size are mainly used for cell purification and separation, and membranes formed on the inner wall of the container are used for culturing.
Membranes coated with chromatographic carriers are suitable for separation and purification.

The physical properties of the surface of a polymer compound, particularly the amount of electric charge, are essential design factors in controlling the adsorption ability with cells. The amount of surface charge can be designed by using condensable or polymerizable monomers, surfactants, polymerization initiators, or other additives that form the interface. Preferably, to provide a negative charge amount, an anionic functional group monomer and an anionic surfactant are used, while to provide a positive charge amount, a cationic functional group monomer, Cationic surfactants are used in combination with appropriate zwitterionic compounds.

Although the mechanism by which cell adsorption ability is controlled by the surface charge and zwitterionic groups of polymer compounds is basically unknown, This is thought to be related to the uneven distribution of The amount of charge on the surface of the polymer compound ranges from -75 to +7
If the voltage is outside the range of 5 mV, the attack on the cells on the surface of the polymer compound will be too strong, resulting in destruction of the cells, which is not preferable. Desirably from -60 to +60 m from the viewpoint of affinity for cells.
It is better to be within the range of V.

The amount of surface charge can be easily measured as the zeta potential.

In the case of particles, the Zeter potential is measured by electrophoresis, which is a phenomenon in which solid particles move through a fixed liquid, and in the case of membranes, it is obtained by using a pressurized liquid in which a solid is fixed. It can be determined by a method of measuring the flowing potential generated by passing a liquid through the solid, or by a method of applying a voltage to both ends of a fixed solid and measuring the mobility of a liquid generated by electroosmosis.

The particle surface zeta potential can be easily determined using Penchem's Laser Zea Model 501, which uses a rotating prism. Regarding the surface zeta potential of the membrane, G
With a device using Ortner's streaming potential measurement cell,
The most suitable method is to determine the zeta potential by measuring the streaming potential.

The cultivation of cells in the presence of the zwitterionic polymer compound according to the present invention can be carried out using the medium used in ordinary cell culture methods, such as liquid culture medium used in shaking culture methods, agitation culture methods, etc., and solids containing agar, etc. The polymer compound is dispersed in a medium in the form of particles, fibers, or films. Also,
It can also be carried out using a culture substrate coated with the polymer compound. Since the polymer compound has a zwitterionic group, it exhibits a pH13 neutralizing effect, and has a surface similar to the cell surface, so it has high affinity for cells. In addition, the cell periphery is stabilized by the cell adsorption effect derived from the surface charge amount and the zwitterionic group. Therefore, culturing cells in the presence of the polymer compound has the advantage that it can adapt to changes in the physical or chemical environment compared to culturing only cells.

Separation or purification of cells in the presence of the polymer compound obtained according to the present invention can be carried out by using a substrate coated with the polymer compound or a plurality of cell suspensions having different properties on the polymer compound. After adsorbing a turbid liquid or a cell suspension containing impurities, the pH is adjusted by taking advantage of the differences in the properties of cells, etc.
This is done by chemical methods such as changes or physical methods such as vibrations. Preferably, the adsorption ability of the polymer compound is controlled to create a difference in adsorption power between a plurality of cells, etc., and then the cells, etc. are separated. The polymer compound has a zwitterionic group, so it exhibits a pH buffering effect, and has a surface similar to the cell surface, so it has high affinity for cells.

Therefore, separation or purification of cells in the presence of the polymer compound has the advantage that cells stably exist.

Examples and comparative examples of the present invention are shown below. In Examples and Comparative Examples, "parts" and "%" are by weight.

Example 1 Bishydroxyethyl taurine 1 was added to a 21 Kolben equipped with an Aion A stirrer, a nitrogen introduction pipe, a temperature control device, a condenser, and a decanter.
34 parts, neopentyl glycol 130 parts, azelaic acid 236 parts, phthalic anhydride 186 parts and xylene 27 parts
Prepare the ingredients and raise the temperature. Water produced by the reaction is azeotroped with xylene and removed.

The temperature was raised to 190°C over about 2 hours from the start of refluxing, stirring and dehydration were continued until the acid value equivalent to carboxylic acid reached 145, and then the mixture was cooled to 140°C.

Then, the temperature was maintained at 140°C, and F Cardura E10
314 parts of persatic acid glycidyl ester (manufactured by Shell) were added dropwise over 30 minutes, and stirring was continued for 2 hours to complete the reaction. The obtained polyester resin having zwitterionic groups had an acid value of 59, a hydroxyl value of 90, and an M
It was n1054.

Example 2 25 parts of taurine, 8 parts of sodium hydroxide, 100 parts of deionized water, and 200 parts of ethylene glycol monoethyl ether were charged into 21 flasks equipped with a stirrer, a cooling tube, and a temperature control device. While stirring, raise the temperature to 100℃. After the contents became uniformly dissolved, Epikote 828 (manufactured by Shell Chemical Co., diglycidyl ether type epoxy resin of bisphenol A, epoxy equivalent: 1
90) A solution consisting of 190 parts and 200 parts of ethylene glycol monoethyl ether is added dropwise over 2 hours. After dripping 5
Stirring and heating are continued for a period of time to complete the reaction.

The reaction solution is acidified with hydrochloric acid, and the resulting precipitate is collected, purified by reprecipitation with ethylene glycol monoethyl ether and water, and dried under reduced pressure to obtain 205 parts of an epoxy resin having an amphoteric ionic group.

The acid value of this resin by KOH titration was 48.6, and the fluorescent
The sulfur content by line analysis was 3%.

Example 3 200 parts of deionized water was placed in a reaction vessel of 11 equipped with a stirrer, a cooler, and a temperature control device, and the temperature was maintained at 80° C. while stirring, and the mixture obtained in Example 1 was added. An aqueous solution consisting of 2 parts of a polyester resin having a zwitterionic group, 3 parts of azobiscyanovaleric acid, 3.1 parts of dimethylaminoethanolamine and 100 parts of deionized water, and 60 parts of methyl methacrylate.
parts, n-butyl acrylate 68 parts, styrene 56 parts,
A liquid mixture consisting of 4 parts of 2-hydroxyethyl acrylate and 12 parts of ethylene glycol dimethacrylate was added dropwise over a period of 120 minutes. After dropping, further heat at 80℃ for 60 minutes.
Stirring was continued for a minute to obtain a resin particle dispersion having a nonvolatile content of 40% and a particle size of 1.5 μm and having an amphoteric ionic group on the particle surface.

After repeating the method of separating the resin particle suspension into a supernatant liquid and resin particles by allowing natural sedimentation and dispersing only the resin particles in deionized water three times, mannitol 0.7 M and magnesium chloride 5 It was dispersed in a washing solution consisting of n+M, centrifuged at 11,000 rpm, dispersed again in the washing solution, and this operation was repeated three times for further washing.

The resin particle dispersion was heated in an autoclave at 120°C, latm.
, treated for 15 minutes to sterilize.

The zeta potential of the resin particles was +5 mV.

Example 4 800 parts of deionized water, 2 parts of polyvinyl alcohol with an average molecular weight of 1500, and 0.02 parts of Amhitol 24B (manufactured by Kao Soap ■, lauryl betaine) were placed in a 1 liter reaction vessel equipped with a stirrer, a cooler, and a temperature control device. Prepare, 40
Dissolve while stirring at 0 rpm and maintaining the temperature at 70°C. To this were added 38 parts of methyl methacrylate, 2 parts of ethylene glycol dimethacrylate, 0.2 parts of 2,2°-azobisisobutyronitrile and 2,2°-azobis-2,
A solution consisting of 0.4 part of 4-dimethylvaleronitrile,
A solution consisting of 5 parts of methanol and 1 part of N-(3-sulfopropyl)-N-methacryloxyethyl-N,N-dimethylammonium betaine was added dropwise over 30 minutes.
After holding for a period of time, the temperature of the system was raised to 80° C. and held for an additional 3 hours to obtain resin particles having amphoteric ionic groups.

The particle size of the resin particles was 75μ.

After repeating the method of separating the resin particle suspension into a supernatant liquid and resin particles by allowing natural sedimentation and dispersing only the resin particles in deionized water three times, mannitol 0.7 M and magnesium chloride 5 It was dispersed in a washing solution consisting of mM, centrifuged at 11,000 rpm, dispersed again in the washing solution, and this operation was repeated three times for further washing. The resin particle dispersion was heated to 120℃ in an autoclave at 1atn+.
, treated for 15 minutes to sterilize.

The zeta potential of the resin particles was -15 mV.

Example 5 A stainless steel beaker was charged with 100 parts of the epoxy resin having amphoteric ionic groups obtained in Example 2, 8 parts of dimethylethanolamine, and 150 parts of deionized water. Manufactured by Anamide, trade name: Cymel 303)
15 parts and 5 parts of ethylene glycol monobutyl ether were gradually added to obtain a resin composition having a zwitterionic group.

This composition was coated on the inside of a glass Petri dish with a dry film thickness of 10
A glass Petri dish coated with a resin composition having a zwitterionic group could be obtained by coating the resin composition so as to have a zwitterionic group and baking it at 170° C. for 20 minutes.

The zeta potential of the resin film was -20 mV.

Example 6 Benzyl adenine (1
0-8M), naphthyl acetic acid (10-5M), and a liquid medium (50d) containing resin particles synthesized according to Example 3 (20mM).
Culture was carried out with rotational shaking at 120 rpm. After 10 days, the cells were collected using a 62 μ nylon filter and washed with sterile water to obtain cultured cells (15 g).

Comparative Example 1 Benzyl adenine (1
Hanakirin cultured cells (2 g) were transferred to a liquid medium (50 d) containing 0-"M), naphthylacetic acid (10-5 M), and morpholine ethanesulfonic acid (20 mM), and
Culture was carried out with rotational shaking at 20 rpm.

After 10 days, the cells were collected using a 62μ nylon filter and washed with sterile water to obtain cultured cells (10g).

Example 7 Hanakirin cultured cells obtained in Example 6 (15 g)
is dispersed in an enzyme solution (100d) having the following composition,
The reaction was carried out at 25° C. with rotational shaking at 14 Orpm. As an enzyme solution, Cellulase Onozuka R5 (2% w
/v, manufactured by Yakult Chemical Industry), Macerozyme R-10 (
1%-/v, manufactured by Yakult Chemical Industry), mannitol (0
, 7M) and adjusted the pH to 5°6, then the pH was adjusted to 0.22
The resin particles synthesized in Example 3 (20mM) were mixed with the mixture that had been passed through a mμ membrane filter for sterilization and used. After culturing for 3 hours, the reaction solution was filtered through a 62μ nylon filter and centrifuged at 1000 rpm for 2 minutes to precipitate protoplasts. The supernatant liquid was removed, and the washing liquid (mannitol 0.7M, magnesium chloride 5M
After adding mM (consisting of resin particles 201 synthesized according to Example 3), centrifugation was performed under the same conditions to obtain Example 4.
Pour the obtained resin particles into a glass column packed with
Protoplasts etc. were adsorbed. When the washing solution was passed through, the cell debris came out first, followed by the protoplasts. The resulting prodoplast effluent was concentrated by centrifugation. In the end, Thoma's hemocytometer determined that 107 cells/
A total of 108 protoplasts were obtained. The zeta potential of this protoplast is measured to be -35
It was mV.

Comparative Example 2 Hanakirin cultured cells obtained in Example 6 (15 g)
was dispersed in an enzyme solution (100 pieces) having the following composition, and reacted at 25° C. with rotational shaking at 14 Orpm. As an enzyme solution, Cellulase Onozuka RS (2
%masu/v+manufactured by Yakult Chemical Industry), Macerozyme R-1
0<t%-/vt manufactured by Yakult Chemical Industry Co., Ltd.) and mannitol (0.7M), and after adjusting the pH to 5°6 with 1M,
．． The mixture was passed through a 22 mμ membrane filter for sterilization, and morpholine ethanesulfonic acid (20 mM) was mixed therein for use. After culturing for 3 hours, the reaction mixture was filtered through a 6-2μ nylon filter and centrifuged at 10,100 rpm to precipitate protoplasts. After removing the supernatant and adding a washing solution (consisting of 0.7M mannitol, 5111M magnesium chloride, and 20mM morpholine ethanesulfone M), the mixture was further centrifuged under the same conditions and washed repeatedly with the washing solution. Finally, 108 protoplasts were obtained using Thoma's hemocytometer (107 cells/total amount requested). However, this protoplast solution contained many cell residues and could not be used for subsequent experiments.

Example 8 In the 6 cm diameter glass petri dish obtained in Example 5,
Benzyl adenine (1
0-'! Hanakirin culture cells (
0.5 g) was transferred and cultured statically.

After 10 days, the cells were collected using a 62μ nylon filter and washed with sterile water to obtain cultured cells (4 g).

Comparative Example 3 Benzyl adenine (10-”M
) and naphthylacetic acid (10-5M) in a liquid medium (10d) containing cultured Hanakirin cells (0.5M).
g) was transferred and cultured statically. After 10 days, the cultured cells (1g
) was obtained.

Claims (12)

[Claims] (1) Has a zwitterionic group and has a surface Jeter potential of -7
A biocompatible biomedical material consisting of a polymeric material with a voltage of 5 to +75 mV. (2) The biomedical material according to item 1, which has a surface Jeter potential of -60 to +60 mV. (3) The biomedical material according to item 1 or 2, which is in the form of particles with an average particle size of 0.01 to 1000 μm. (4) The biomedical material according to item 1 or 2, which is in the form of a membrane. (5) The biomedical material according to item 1 or 2, wherein the polymeric material is obtained by polymerizing an ethylenically unsaturated monomer in the presence of a zwitterionic compound. (6) The biomedical material according to item 5, wherein the ratio of the zwitterionic compound to the ethylenically unsaturated monomer is 1:99 to 50:50. (7) The biomedical material according to item 1 or 2, wherein the polymer material is obtained by chemically bonding or blending a zwitterionic compound to a substrate polymer compound. (8) The biomedical material according to item 7, wherein the ratio of the zwitterionic compound to the matrix polymer compound is 1:99 to 50:50. (9) Has a zwitterionic group and has a surface Jeter potential of -7
A cell processing method characterized by using a biomedical material made of a polymeric material having a voltage of 5 to +75 mV as a cell adsorbent. (10) The method of item 9, wherein the treatment is cell culture. (11) The method according to item 9, wherein the treatment is cell separation. (12) The method of item 9, wherein the treatment is cell purification.