CELL CHIP

Abstract

A cell chip is provided that exhibits excellent resistance to cell desiccation, cell retention, and detection of chemical substances such as odorants. The cell chip comprises a compartment containing a cell and a gel that can be converted into a sol.

Description

TECHNICAL AREA

[0001] The present invention relates to a cell chip and the like. STATE OF THE ART

[0002] Groups of odorants have been identified that are associated with specific human diseases and mental states. Due to their high value as diagnostic markers, there is growing interest in developing various odor sensors that target these substances. Biological odorant receptors exhibit excellent properties, including diversity, sensitivity, and selectivity, which are not found in conventional odorant sensor elements such as semiconductors. Therefore, these odorant receptors offer a promising basis for the development of novel odor sensors using them as sensor elements.

[0003] PTL 1 discloses the use of cells expressing modified odor receptors or of lipid bilayer membranes containing modified odor receptors as odor sensors. CITATION LIST Patent Literature

[0004] PTL 1: WO2022 / 024902A SUMMARY OF THE INVENTION Technical Problem

[0005] Odor sensors based on the artificial fabrication of lipid bilayer membranes equipped with sensor proteins (e.g., olfactory receptors) are often inefficient to manufacture, and there is a need to further improve the manufacturing efficiency of odor sensors. The present inventors therefore focused on the use of cells expressing sensor proteins.

[0006] From a user-friendliness perspective, cells used as chemical sensors, for example as odor sensors, are preferably those that are pre-produced and stored in containers so they can be used as needed, rather than cells that have to be cultured and produced each time they are used. In the former case, it is necessary to protect the cells from drying out until they are needed, and it is also important that the cells remain stable, taking into account factors such as transport from the manufacturer to the point of use.

[0007] Furthermore, from the perspective of using the cells for disease diagnosis and other assessments, it is desirable to use several types of cells expressing different olfactory receptors simultaneously. From this perspective, using the cells in the form of a cell chip is preferable.

[0008] Accordingly, an objective of the present disclosure is to provide a cell chip that exhibits excellent cell desiccation resistance, cell retention and detection of chemical substances, such as odorants. Solution to the problem

[0009] In the course of their research, the present inventors focused on the fact that a cell chip containing compartments, each containing cells and a hydrogel, can prevent the cells from drying out and keep them stable. However, they found that the increase in the sensor protein's reactivity after adding a chemical substance in the presence of a hydrogel occurs gradually, which impairs rapid and stable measurement of the activity. Based on this finding, the inventors discovered that a cell chip containing compartments, each containing cells and a gel that can be converted into a sol, can achieve the above-described goal and solve the above-described problem. Specifically, the present disclosure includes the following embodiments. Point 1.

[0010] Cell chip, comprehensive a compartment containing a cell and a gel that can be converted into a sol. Point 2.

[0011] Cell chip according to point 1, wherein the gel is a gel which can be converted into a sol by temperature control, light irradiation or the addition of a chemical agent. Point 3.

[0012] Cell chip according to point 2, wherein the gel is a gel that can be converted into a sol by temperature control. Point 4.

[0013] Cell chip according to point 3, wherein the gel has a sol formation temperature of 40 °C or less. Point 5.

[0014] Cell chip according to point 3, wherein the gel has a sol formation temperature of 30 °C or less. Point 6.

[0015] Cell chip according to point 3, wherein the gel has a sol-gel transition temperature of 4 to 25 °C. Point 7.

[0016] Cell chip according to point 1, wherein the sol formed after solation of the gel has a viscosity of 20 mPa·s or less. Point 8.

[0017] Cell chip according to point 1, wherein the gel has a storage elasticity modulus of 0.01 to 20 kPa at 4 °C. Point 9.

[0018] Cell chip according to at least one of points 1 to 8 for use in a method for detecting a chemical test substance, wherein the method comprises bringing the cell into contact with the chemical test substance after the solvent has been dissolved. Point 10.

[0019] Cell chip according to at least one of points 1 to 8 for use in detecting light from the cell. Item 11.

[0020] Cell chip according to at least one of points 1 to 8, wherein the cell is an insect cell. Item 12.

[0021] Cell chip according to at least one of points 1 to 8, wherein the cell comprises an exogenous polynucleotide containing a coding sequence for a sensor protein. Point 13.

[0022] Cell chip according to point 12, wherein the sensor protein is an odor receptor protein. Item 14.

[0023] Kit for immobilizing a cell, comprising a component for forming a gel that can be converted into a sol. Point 15.

[0024] Kit according to point 14 for immobilizing a cell onto a cell chip. Item 16.

[0025] Method for producing a cell for transport, comprising immobilizing a cell with a gel that can be converted into a sol. Item 17.

[0026] Method for detecting light from a cell, comprising the conversion of a gel into a sol, wherein the gel can immobilize the cell and be converted into a sol. Item 18.

[0027] Gel that can be converted into a sol, where the gel can be converted into a sol by temperature control the gel has a sol formation temperature of 40 °C or less, the gel has a sol-gel transition temperature of 4 to 25 °C, the sol formed after solation of the gel has a viscosity of 20 mPa·s or less and The gel has a storage elasticity modulus of 0.01 to 20 kPa at 4 °C. Item 19.

[0028] Sol for the formation of the gel according to point 18. Item 20.

[0029] Kit as per point 14, comprising the gel as per point 18 and / or the sol as per point 19. Point 21.

[0030] Kit according to point 20 for immobilizing a cell onto a cell chip. Item 22.

[0031] Procedure according to point 16, wherein the gel is the gel according to point 18. Item 23.

[0032] Procedure according to point 17, wherein the gel is the gel according to point 18. Advantageous effects of the invention

[0033] The present disclosure relates to a cell chip exhibiting excellent resistance to cell desiccation, cell retention, and detection of chemical substances, such as odorants. Furthermore, the present disclosure also relates to a kit for immobilizing cells onto a cell chip, a method for producing cells for transport, a method for detecting light from cells, a gel suitable for use in the cell chip, a sol for forming such a gel, and so on. BRIEF DESCRIPTION OF THE DRAWINGS Fig.Figure 1 shows the measurement results of the activity of an olfactory receptor in test example 4. The vertical axis shows the fluorescence intensity, and the horizontal axis shows the time course. The arrow indicates the time at which compound a (a substance to which ORA cells respond) was added. Fig. Figure 2 shows the measurement results of the activity of an olfactory receptor in test example 5. The vertical axis shows the fluorescence intensity, and the horizontal axis shows the time course. The arrow indicates the time at which compound a (a substance to which ORA cells respond) was added. Fig.Figure 3 shows the measurement results of the activity of an olfactory receptor in test example 6. The vertical axis shows the value determined by subtracting the mean background value (the average of the background values ​​during the 20 seconds before the addition of the compound) from the maximum fluorescence intensity. On the horizontal axis, the percentage indicates the gelatin concentration, and "No Gel" indicates the case where a buffer solution was added instead of a gelatin solution. Fig.Figure 4 shows the measurement results of the activity of an olfactory receptor in test example 7. The vertical axis indicates the value determined by subtracting the mean background value (the average of the background values ​​during the 20 seconds before the addition of the compound) from the maximum fluorescence intensity. On the horizontal axis, "No Gel" denotes the case where a buffer solution was added instead of a gelatin solution, while the other cases indicate the presence or absence of sol after liquefaction of a hydrogel. Fig. Figure 5 shows the viscosity measurement results of a sol in test example 8. The vertical axis indicates the measured viscosity values. The horizontal axis indicates the fish gelatin concentration in a gelatin solution. Fig.Figure 6 shows the viscosity measurement results of a sol in test example 9. The vertical axis indicates the measured viscosity values. The horizontal axis indicates the fish gelatin concentration in a gelatin solution. Fig. Figure 7 shows the measurement results of the storage elasticity modulus and the sol-gel transition temperature in test example 10. DESCRIPTION OF PERFORMANCE FORMS

[0034] In the present description, the terms “comprehensive”, “containing” and “inclusive” encompass the concepts of encompassing, containing, essentially consisting of and comprising.

[0035] In one embodiment, the present disclosure relates to a cell chip comprising a compartment containing a cell and a gel that can be converted into a sol (hereinafter referred to as "the cell chip of the present disclosure"). The cell chip is explained below.

[0036] The cell is not particularly limited. From the perspective of suitability for chemical detection, animal cells, such as insect or mammalian cells, are preferable, with insect cells being particularly favored due to their ease of handling, as they do not require, for example, CO2 or temperature control.

[0037] Examples of usable insect cells include Sf cells, MG1 cells, High Five™ cells, and BmN cells. Usable Sf cells include, for example, Sf9 cells (ATCC CRL1711) and Sf21 cells. Insect cells that possess coding sequences for insect olfactory receptors in their genome can be used as chemical sensors. Of these, insect cells from insects of the family Arctiidae are particularly preferred.

[0038] The cells derived from insects of the family Arctiidae are not particularly restricted, as long as they are primarily cultured cells or established cell lines of biological components derived from insects of the family Arctiidae.

[0039] Examples of the family Arctiidae include subfamilies such as Arctiinae, Lithosiinae, and Syntominae, with the subfamily Arctiinae being preferred. Within the subfamily Arctiinae, for example, the genera Spilosoma, Spilarctia, or Rhagonis are preferred, with the genus Spilosoma being particularly favored. Although there is no specific restriction regarding the genus Spilosoma, Spilosoma imparilis is especially favored.

[0040] Cells derived from insects of the family Arctiidae can be obtained from known biological banks or can be collected and cultured from live insects of the family Arctiidae according to or with reference to known procedures; if necessary, they can be established as cell lines.

[0041] Examples of cells derived from Spilosoma imparilis are FFPRI-SpIm-2AM-SF cells (MAFF No.: 275052) and FFPRI-SpIm-2AM-IPL411 cells (MAFF No.: 275053) from the National Agriculture and Food Research Organization's gene bank project.

[0042] The cell preferably contains an exogenous polynucleotide that carries a coding sequence for a sensor protein. This enables the expression of any sensor protein and allows for increased expression levels of a target sensor protein, thereby improving the detection sensitivity for a target chemical.

[0043] The “exogenous polynucleotide” is not particularly restricted as long as it is a polynucleotide containing a base sequence that does not originate from the genomic DNA (especially the chromosomal genomic DNA) of an insect cell.

[0044] In this description, the polynucleotide includes not only typical polynucleotides such as DNA and RNA found in organisms, but also polynucleotides with known chemical modifications, artificial polynucleotides, and similar polynucleotides, as listed below. To prevent degradation by hydrolases such as nucleases, the phosphate group of each nucleotide can be replaced by a chemically modified phosphate group such as phosphorus thioate (PS), methylphosphonate, or phosphorus dithionate. The hydroxyl group at position 2 of the ribose of each ribonucleotide can also be substituted by -OR (R denoting, for example, CH3(2'-O-Me), CH2CH2OCH3(2'-O-MOE), CH2CH2NHC(NH)NH2, CH2CONHCH3, or CH2CH2CN).Additionally, the nucleobase unit (pyrimidine, purine) can be chemically modified, for example by introducing a methyl group or a cationic functional group at position 5 of the pyrimidine base or by substituting the carbonyl group at position 2 with a thiocarbonyl group. Furthermore, the polynucleotide of the present invention also includes, but is not limited to, those formed by modifying the phosphate unit or the hydroxyl unit, for example with biotin, an amino group, a lower alkylamino group, or an acetyl group. The polynucleotide to be used can, for example, also be BNA (LNA), which is produced by crosslinking the 2' oxygen and the 4' carbon in the ribose unit of a nucleotide to fix the ribose unit in the N-conformation.

[0045] The sensor protein can be selected from proteins capable of detecting the presence of a chemical substance, such as a receptor protein that uses a chemical substance as a ligand. The sensor protein is preferably an olfactory receptor protein.

[0046] The insect olfactory receptor protein is a membrane protein with a seven-transmembrane structure and functions as an odor sensor in organisms. The olfactory receptor protein is composed of the following components, linked sequentially from the amino terminus (hereinafter referred to as the "N-terminus") to the carboxyl terminus (hereinafter referred to as the "C-terminus"): N-terminal region (NT), first transmembrane domain (TM1), first extracellular loop (EC1), second transmembrane domain (TM2), first intracellular loop (IC1), third transmembrane domain (TM3), second extracellular loop (EC2), fourth transmembrane domain (TM4), second intracellular loop (IC2), fifth transmembrane domain (TM5), third extracellular loop (EC3), sixth transmembrane domain (TM6), third intracellular loop (IC3), seventh transmembrane domain (TM7), and C-terminal region (CT). In the present disclosure, each area is described by structure prediction using TMPred (K. Hofmann, W.Stoffel, TMbase - a database with segments of membrane-spanning proteins, Biol. Chem. Hoppe-Seyler, 374 (1993), p. 166, https: / / embnet.vital-it.ch / software / TMPRED_form.html) (with standard conditions).

[0047] The source insect for the insect olfactory receptor protein is preferably an insect from the order Diptera, such as from the families Culicidae and Drosophilidae; an insect from the order Lepidoptera, such as from the family Bombycidae; an insect from the order Hymenoptera, such as from the family Apidae; an insect from the order Orthoptera, such as from the family Acrididae; and an insect from the order Hemiptera, such as from the family Cimicidae. Examples of insects from the family Culicidae include Anopheles gambiae, Aedes aegypti, and Culex quinquefasciatus.Examples of insects from the family Drosophilidae include Drosophila melanogaster, Drosophila pseudoobscura, and Drosophila virilis. Examples of insects from the family Bombycidae include Bombyx mori, Bombyx mandarina, and Trilocha varians. Examples of insects from the family Apidae include Apis mellifera, Apis florea, Apis dorsata, and Bombus terrestris. Examples of insects from the family Acrididae include Locusta migratoria. Examples of insects from the family Cimicidae include Cimex lectularius.

[0048] Examples of wild-type insect olfactory receptor proteins include, in particular, the following: AaOR1, AaOR2, AaOR4, AaOR5, AaOR6, AaOR8, AaOR9, AaOR10a, AaOR15, AaOR22, AaOR24, AaOR25, AaOR26, AaOR27, AaOR28, AaOR30, AaOR34, AaOR36, AaOR38, AaOR41a, AaOR41b, AaOR42, AaOR43, AaOR44, AaOR47, AaOR49, AaOR50, AaOR52, AaOR54, AaOR58, AaOR59, AaOR60, AaOR61, AaOR64, AaOR65, AaOR66, AaOR67a AaOR69a, AaOR70, AaOR71, AaOR72a, AaOR73, AaOR74, AaOR75, AaOR77, AaOR78, AaOR79, AaOR81, AaOR83b, AaOR84, AaOR85, AaOR86, AaOR87, AaOR91, AaOR95, AaOR97, AaOR96, AaOR99, AaOR100, AaOR102, AaOR103, AaOR104a, AaOR105, AaOR107, AaOR108, AaOR109, AaOR110, AaOR112, AaOR114, AaOR116, AaOR117, AaOR118, AaOR122, AaOR125, AaOR128, AgOR1, AgOR2, AgOR3, AgOR4, AgOR5, AgOR6, AgOR8, AgOR9, AgOR10, AgOR11a, AgOR12a, AgOR12b, AgOR13, AgOR14, AgOR15, AgOR16a, AgOR17, AgOR18, AgOR20, AgOR21, AgOR23, AgOR25, AgOR26, AgOR27, AgOR28, AgOR30, AgOR34, AgOR36, AgOR37, AgOR38, AgOR39a, AgOR40, AgOR42, AgOR44, AgOR45, AgOR46, AgOR47,AgOR49, AgOR50, AgOR54, AgOR56a, AgOR57, AgOR60, AgOR61, AgOR62, AgOR63, AgOR64, AgOR65, AgOR69, AgOR70, AgOR71, AgOR72, AgOR74, AgOR75, AgOR76a, AmOR1, AmOR3, AmOR9, AmOR10, AmOR13, AmOR41, AmOR51, AmOR52, AmOR55, AmOR71, AmOR73, AmOR78, AmOR85, AmOR89, AmOR90, AmOR114, AmOR115, AmOR118, AmOR120, AmOR121, AmOR161, BmOR1, BmOR2, BmOR3, BmOR4, BmOR5, BmOR8, BmOR9, BmOR10, BmOR13, BmOR17, BmOR18, BmOR23, BmOR24, BmOR25, BmOR35, BmOR36, BmOR42, BmOR45, BmOR49, BmOR51, BmOR52, BmOR55, BmOR56, BmOR61, DmOR1a, DmOR9a, DmOR19a, DmOR22a, DmOR22b, DmOR22c, DmOR24a, DmOR30a, DmOR33a, DmOR33b, DmOR33c, DmOR35a, DmOR42b, DmOR43a, DmOR45a, DmOR45b, DmOR47a, DmOR49b, DmOR59b, DmOR65b, DmOR65c, DmOR67b, DmOR67c, DmOR69a, DmOR71a, DmOR74a, DmOR82a, DmOR83a, DmOR83c, DmOR85a, DmOR85c, DmOR85e, DmOR85f, DmOR88a, DmOR92a, DmOR94a, DmOR94b and DmOR98b.

[0049] In this description, GR denotes "olfactory receptor." Dm represents the Drosophila melanogaster derivative, Bm the Bombyx mori derivative, Ag the Anopheles gambiae derivative, and Aa the Aedes aegypti derivative. The amino acid sequences of various olfactory receptor proteins, including this one, and their coding sequences are either known or can be readily identified by sequence identity searches based on known sequences.

[0050] The sensor protein may contain one or more amino acid mutations in its wild-type amino acid sequence, provided that the chemical reactivity is not significantly reduced. The term "not significantly reduced" means, for example, that the chemical reactivity of a sensor protein containing amino acid mutations is, for example, at least 50%, preferably at least 60%, more preferably at least 70%, more preferably at least 80%, and more preferably at least 90% of the chemical reactivity of the wild-type sensor protein, which is assumed to be 100%.

[0051] An amino acid mutation is, for example, a substitution, insertion, addition or deletion of an amino acid, preferably a substitution and particularly preferably a conservative substitution.

[0052] In this description, "conservative substitution" refers to the substitution of one amino acid residue by another amino acid residue with a similar side chain. For example, substitution between amino acids with a basic side chain, such as lysine, arginine, or histidine, is considered a conservative substitution.The following substitutions between other amino acid residues are also considered conservative substitutions: the substitution between amino acid residues with an acidic side chain, such as aspartic acid and glutamic acid; the substitution between amino acid residues with an uncharged polar side chain, such as glycine, asparagine, glutamine, serine, threonine, tyrosine, or cysteine; the substitution between amino acid residues with a nonpolar side chain, such as alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, or tryptophan; the substitution between amino acid residues with a β-branched side chain, such as threonine, valine, or isoleucine; and the substitution between amino acid residues with an aromatic side chain, such as tyrosine, phenylalanine, tryptophan, or histidine.

[0053] The sensor protein can contain its wild-type amino acid sequence or an amino acid sequence which, for example, has at least 70%, preferably at least 80%, more preferably at least 90%, more preferably at least 95%, more preferably at least 98% and most preferably at least 99% identity with the wild-type amino acid sequence.

[0054] In this description, the "identity" of amino acid sequences refers to the degree to which two or more comparable amino acid sequences are identical. Thus, the higher the degree of similarity between two amino acid sequences, the higher the identity or similarity of these sequences. The degree of amino acid sequence identity can be determined, for example, using FASTA, a sequence analysis tool, with standard parameters. Alternatively, the degree of amino acid sequence identity can be determined using the BLAST algorithm by Karlin and Altschul (Karlin S., Altschul SF Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes, Proc. Natl. Acad. Sci. USA. 87: 2264-2268 (1990); Karlin S., Altschul SF Applications and statistics for multiple high-scoring segments in molecular sequences, Proc. Natl. Acad. Sci. USA. 90: 5873-7 (1993)).Based on this BLAST algorithm, a program called "BLASTX" was developed. The specific techniques of these analysis methods are known and can be found on the website of the National Center for Biotechnology Information (NCBI) (http: / / www.ncbi.nlm.nih.gov / ).

[0055] The sensor protein may contain other amino acid sequences, such as protein tags, fluorescent proteins, luminescent proteins, signal sequences, or other proteins or peptides bound to it, provided that the chemical reaction activity is not significantly impaired. Examples of protein tags include biotin, His tag, FLAG tag, Halo tag, MBP tag, HA tag, Myc tag, V5 tag, and PA tag.

[0056] In this description, "chemical reaction activity" refers to the ability of a sensor protein to recognize a chemical substance and, either alone or in conjunction with other proteins, to trigger signal transduction activity (e.g., ion channel activity). In the case of olfactory receptors, "chemical reaction activity" refers to the ability of an olfactory receptor to recognize a chemical substance and, together with an olfactory receptor co-receptor, to form an olfactory receptor complex, thereby exhibiting ion channel activity upon activation. The chemical reaction activity of a sensor protein can be measured by using the signal transduction activity of the sensor protein in contact with a chemical substance as an indicator (e.g., by quantifying and evaluating the amount of signaling molecules).In the case of olfactory receptors, their chemical reaction activity can be measured using the ion channel activity of an olfactory receptor complex, formed from an olfactory receptor in contact with a chemical substance and an olfactory receptor co-receptor, as an indicator. For example, a chemical substance is brought into contact with cells expressing a protein that becomes fluorescent or luminescent in response to the influx of ions (e.g., calcium ions) into the cells when (a) an olfactory receptor, (b) an olfactory receptor co-receptor, and (c) an olfactory receptor complex react, and then the amount of luminescence from the cells is measured. The greater the measured amount of luminescence, the higher the reaction activity of the olfactory receptor to the chemical substance is determined to be. In particular, the chemical reaction activity can be measured according to the procedure described in PTL 1.

[0057] The coding sequence for the sensor protein is not particularly restricted, as long as it is a base sequence that codes for the sensor protein. In one embodiment, the exogenous polynucleotide contains an expression cassette for a sensor protein. The expression cassette is not particularly restricted as long as it is a polynucleotide capable of expressing the sensor protein within a cell. A typical example of an expression cassette for a sensor protein is a polynucleotide containing a promoter and the coding sequence for the sensor protein, which is under the control of the promoter.

[0058] The promoter is not particularly restricted and can be selected accordingly. For example, the promoter to be used can be chosen from various types of Pol II promoters. Examples of Pol II promoters include, but are not limited to, the CMV promoter, the EF1 promoter, the SV40 promoter, the MSCV promoter, and promoters derived from insect genes.

[0059] If the sensor protein is an insect olfactory receptor, it is preferable that the exogenous polynucleotide contains a coding sequence for an insect olfactory receptor co-receptor. The insect olfactory receptor co-receptor is a membrane protein with a seven-transmembrane structure similar to the olfactory receptor and functions by forming a heterocomplex with an olfactory receptor. The olfactory receptor complex, which is a heterocomplex consisting of an olfactory receptor and an olfactory receptor co-receptor, exhibits ion channel activity that is activated by odorants. Upon activation, the olfactory receptor complex allows the influx of cations such as sodium ions (Na⁻). + ) and calcium ions (Ca 2+ ) into the cells.

[0060] It is preferred that the exogenous polynucleotide contains an encoding sequence for a protein that becomes fluorescent or luminescent in response to the influx of ions (e.g., calcium ions) into the cell when the sensor protein (in particular, the olfactory receptor protein) reacts. Examples of such proteins include aequorin, yellow chameleon (YC), and GCaMP. Alternatively, the cell of the present disclosure preferably contains an ion-dependent fluorescent dye, such as calcium ion-dependent fluorescent dyes (e.g., Fura-2, Fluo-3, and Fluo-4).

[0061] The exogenous polynucleotide preferably contains a coding sequence for a drug-resistant gene to enable drug screening of cells. The drug-resistant gene can be selected from a gene resistant to a drug suitable for cell drug screening. Examples of drug-resistant genes include chloramphenicol resistance, tetracycline resistance, neomycin resistance, erythromycin resistance, spectinomycin resistance, kanamycin resistance, hygromycin resistance, and puromycin resistance.

[0062] It is preferred that the exogenous polynucleotide contains coding sequences such as a coding sequence for an insect olfactory receptor co-receptor, a coding sequence for a chromogenic or luminescent protein, and a coding sequence for a drug resistance gene in the form of an expression cassette. The structure of the expression cassette is similar to that of the expression cassette for the sensor protein. The promoter of the expression cassette can be shared by several coding sequences.

[0063] The exogenous polynucleotide is preferably integrated into genomic DNA (especially chromosomal genomic DNA). This enables stable expression of the sensor protein, making it suitable for chemical detection. In this case, the polynucleotide can be a single contiguous region within the genomic DNA or a combination of two or more contiguous regions (e.g., a form in which the sensor protein coding sequence is contained in contiguous region A and the coding sequence for a drug-resistant gene is contained in contiguous region B, which is separated from contiguous region A, or a form in which the sensor protein coding sequence is contained in both contiguous region A and contiguous region B).

[0064] In another embodiment, the polynucleotide can be in a state where it is not integrated into the genomic DNA. In this case, the exogenous polynucleotide can, for example, be in the form of a vector. The polynucleotide can be a single polynucleotide molecule or two or more polynucleotide molecules (e.g., a form in which the sensor protein coding sequence is contained in polynucleotide molecule A, while the coding sequence for a drug resistance gene is contained in polynucleotide molecule B, which is a separate molecule from polynucleotide molecule A, or a form in which the sensor protein coding sequence is contained in both polynucleotide molecule A and polynucleotide molecule B).

[0065] The gel is not particularly limited as long as it can be converted into a sol. It appears that the solation ensures the high diffusivity of a target substance to be detected and enables fast and stable measurement. Furthermore, the solation simplifies the removal of the gel, allowing it to be replaced by a solution with better diffusivity of the target substance to be detected. Therefore, in one embodiment, the present disclosure relates to a kit for immobilizing a cell on a cell chip, which includes a component for forming a gel that can be converted into a sol.

[0066] Examples of gels include those that can be converted into a sol by temperature control, light irradiation, or the addition of a chemical agent. Of these, gels that can be converted into a sol by temperature control are preferred from the perspective of the ease of preparation of the gel components, the simplicity of the solation, and the reversibility of the sol-gel transition; gels that can be converted into a sol by raising the temperature are even more preferred.

[0067] From the perspective of establishing contact with the cell in the cell chip of the present disclosure in order to immobilize the cell more firmly and at the same time limit thermal damage to the cell during sol formation, the sol formation temperature of the gel, which can be converted into a sol by temperature control, is preferably 40 °C or less, more preferably 35 °C or less, more preferably 30 °C or less, and most preferably 28 °C or less. From the perspective of maintaining the gel stably at a temperature desirable for cell preservation, the lower limit of such a temperature is preferably 10 °C, 15 °C, 20 °C, or 25 °C.

[0068] The “sol formation temperature” can be defined as the temperature at which the loss factor exceeds 1 when the gel is heated at a rate of +1 °C / min in the dynamic viscoelasticity measurement in test example 10 described below.

[0069] The sol-gel transition temperature (i.e., the temperature at which a sol transitions to a gel prior to gel formation) of the gel that can be converted to a sol by temperature control is preferably 35 °C or less, preferably 30 °C or less, more preferably 25 °C or less, and particularly preferably 20 °C or less, in order to achieve contact with the cell in the cell chip of the present disclosure and to immobilize the cell more firmly while limiting thermal damage to the cell during sol production and gel formation.The lower limit of the temperature is not particularly restricted as long as it is 0 °C or higher; however, from the point of view of enabling stable gel formation in a temperature range in which the cells do not freeze, the lower limit of the temperature is preferably 4 °C or higher, and from the point of view of maintaining the gelled state regardless of the temperature increase during transport, the lower limit of the temperature is even more preferably 10 °C or higher.

[0070] The sol-gel transition temperature can be measured according to the procedure described below in test example 10.

[0071] The gel, which can be converted into a sol, preferably forms a sol with a viscosity of 20 mPa·s or less after solventization. To further improve the diffusivity of the target substance to be detected, contribute to faster and more stable measurements, and enhance processability during cell seeding, the viscosity is preferably 15 mPa·s or less, more preferably 10 mPa·s or less, more preferably 8 mPa·s or less, more preferably 6 mPa·s or less, and most preferably 5 mPa·s or less. The lower limit of the viscosity is not particularly restricted and can, for example, be 0 mPa·s, 0.1 mPa·s, 0.2 mPa·s, 0.5 mPa·s, or 1 mPa·s.

[0072] The viscosity can be measured according to or with reference to the procedure described in Test Example 8 or Test Example 9. The value measured by either method can be used as the viscosity. Preferably, the viscosity is a value measured according to or with reference to the procedure described in Test Example 9 (i.e., the viscosity after the sol has been left to stand for 2 days at a sol formation temperature). A relatively low viscosity indicates that the time-dependent increase in viscosity after sol formation is suppressed, demonstrating excellent processing properties.

[0073] The storage elasticity modulus at 4 °C of the gel, which can be converted into a sol, is not particularly limited and is, for example, 0.001 to 100 kPa. From the perspective of preserving cells while simultaneously limiting the external forces acting on the cells in contact with the gel (i.e., limiting damage to the cells), the gel preferably has a storage elasticity modulus at 4 °C of 0.01 to 20 kPa, more preferably of 0.02 to 10 kPa, more preferably of 0.05 to 5 kPa, and more preferably of 0.07 to 2 kPa.

[0074] The storage elasticity modulus can be measured according to the procedure described below in test example 10.

[0075] The component that forms a gel capable of being converted into a sol (gel-forming component: a component that is cross-linked to form a network) is not particularly limited; examples include gelatin, polysaccharides such as agar, carrageenan, starch, and xanthan gum, as well as water-soluble synthetic polymers such as PVA and PEG. Of these, gelatin is a particularly favored component from the standpoint of ease in developing desirable properties. Gelatin is obtained by pretreating crude collagen materials, such as bovine bone, bovine hide, porcine bone, porcine skin, and fish scales, with an acid or alkali, followed by washing with water, hot water extraction, purification, and concentration for drying. The sol and gel properties of gelatin vary depending on the type of crude collagen material, the pretreatment, the hot water extraction conditions, and other factors.In order to achieve the sol temperature, sol-gel transition temperature, sol viscosity and gel elasticity modulus suitable for the present invention, fish-derived gelatin (fish gelatin) is particularly preferred among the various types of gelatin; the sol and gel properties suitable for the present invention can be achieved by appropriately adjusting the pretreatment and the hot water extraction conditions.

[0076] The gel-forming component can be a single type or a combination of two or more types.

[0077] In one embodiment of the present disclosure, the gelatin content in the gel that can be converted into a sol is preferably 50% by mass or more, more preferably 70% by mass or more, more preferably 80% by mass or more, more preferably 85% by mass or more, particularly preferably 90% by mass or more and particularly preferably 95% by mass or more (in particular 100% by mass) based on 100% by mass of the gel-forming component.

[0078] In one embodiment of the present disclosure, the gelatin content in the gel that can be converted into a sol is preferably 50 wt% or more, more preferably 70 wt% or more, more preferably 80 wt% or more, more preferably 85 wt% or more, particularly preferably 90 wt% or more and particularly preferably 95 wt% or more (in particular 100 wt%) based on 100 wt% of the solid content forming the gel.

[0079] If the gel convertible to a sol contains gelatin, the concentration of gelatin in the gel is preferably 0.5 to 10 wt%, more preferably 0.7 to 8 wt%, more preferably 0.8 to 6 wt%, more preferably 0.9 to 4 wt% and particularly preferably 1 to 3.5 wt%, from the point of view of the ease of developing desirable properties.

[0080] The solvent for the gel, which can be converted into a sol, is not particularly restricted as long as it does not significantly impair cell viability. It is preferred that the solvent contains water. The water content of the solvent is preferably 50% by weight or more, more preferably 70% by weight or more, more preferably 80% by weight or more, more preferably 85% by weight or more, particularly preferably 90% by weight or more, and particularly preferably 95% by weight or more (especially 100% by weight).

[0081] In one embodiment, the present disclosure relates to a gel that can be converted into a sol and a sol for forming the gel. In particular, the gel can be converted into a sol by temperature control and fulfills one or more (preferably two, more preferably three, and particularly preferably five) requirements selected from the following group: (a) the gel has a sol formation temperature within a predetermined range, (b) the gel has a sol-gel transition temperature within a predetermined range, (c) the sol formed after solation of the gel has a viscosity within a predetermined range, (d) the gel has a storage elasticity modulus at 4 °C within a predetermined range, and (e) the sol formed after gelation at 4 °C and subsequent resolation by heating to 25 °C has a viscosity within a predetermined range.

[0082] The compartments of the cell chip of the present disclosure preferably each contain cells and a gel that can be converted into a sol.

[0083] The configuration of the compartments is not particularly restricted, as long as they can retain cells and a gel. From the perspectives of cell desiccation resistance, cell retention, manufacturing efficiency, or chemical detection, the compartments are preferably designed in the form of wells.

[0084] The material of the compartments is not particularly restricted, as long as it can retain cells and a gel. The material could be, for example, resin or metal.

[0085] The arrangement of the cells and the gel within the compartment is not particularly restricted, as long as cell desiccation resistance and cell retention are ensured. For example, the cells in each compartment are encapsulated in the gel. The embodiment of the encapsulation is not particularly restricted, as long as it is an embodiment in which at least some of the cells are shielded from the air outside the cell chip by the gel.

[0086] From a detection sensitivity perspective, each compartment typically contains multiple cells. The number of cells per unit area (cm²) 2 ) in a compartment, for example, is 5,000 to 2,000,000 cells / cm². 2 From the perspective of detection sensitivity and cell lifetime, the number of cells per unit area is preferably 10,000 to 1,500,000 cells / cm². 2 , preferably 10,000 to 1,000,000 cells / cm² 2, even more preferred are 20,000 to 1,000,000 cells / cm² 2 , even more preferred are 50,000 to 700,000 cells / cm² 2 and especially preferred are 100,000 to 500,000 cells / cm² 2 .

[0087] From the perspective of detection sensitivity or manufacturing efficiency, the floor area of ​​a single compartment is preferably 0.5 to 100 mm². 2 , even more preferably 1 to 30 mm 2 and most preferred 1.5 to 15 mm 2 In one embodiment of the present invention, the upper limit of the area can be set to 80 mm. 2 , 60 mm 2 or 40 mm 2 be determined.

[0088] From the point of view of detection sensitivity or manufacturing efficiency, the cell chip preferably comprises 10 to 2,000, more preferably 30 to 1,000 and most preferably 50 to 500 compartments.

[0089] It is preferred that the cell chip of the present disclosure comprises two or more (preferably three or more, more preferably four or more, more preferably five or more, ten or more, 15 or more or 20 or more) types of cells which differ from one another with respect to the type of sensor protein.

[0090] The cell chip of the present disclosure can be produced by gelling a sol to form the sol-forming gel in a compartment containing the sol and the cells. The sol can be obtained according to or by reference to a known method. If gelatin is used as the gel-forming component, it is advantageous from the point of view of the simple preparation of a gel / sol with the desirable properties described above to carry out the following procedure: Powdered gelatin is added to a solvent, the mixture is stirred for 10 to 60 minutes (preferably 20 to 40 minutes) at 12 to 30 °C (preferably 15 to 25 °C), then the mixture is heated to the dissolution temperature of the gelatin (e.g., 30 to 60 °C) and the solution is stirred until complete dissolution.

[0091] The cell chip described in this disclosure can be used to detect chemical substances (especially odorants). These chemical substances can be, for example, those present in a sample such as bodily fluids (e.g., urine, blood, and saliva), air (e.g., indoor air and air in packaging), and water (e.g., river water, seawater, tap water, clean water, and wastewater). In this case, the sample is placed in the compartments of the cell chip described in this disclosure, allowing the chemical substance in the sample to reach the cells and come into contact with the sensor protein within the cells. A chemical substance can be detected, for example, by detecting ions flowing into a cell (e.g., by detecting a protein that exhibits color change or luminescence in response to ions).

[0092] As described above, the cell chip of this disclosure enables rapid and stable measurement by converting a gel into a sol. Thus, the cell chip of this disclosure can be used in a method for detecting a chemical test substance, which, after solation of the gel, involves bringing the cells into contact with the chemical test substance. After solation, the chemical test substance can be brought into contact with the cells in the presence of the sol; alternatively, the sol can be removed and, optionally, another liquid added to the compartments, after which the chemical substance to be tested can be brought into contact with the cells. When detecting luminescence from the cells (luminescence due to the addition of a substrate such as coelenterazine), the problem of not being able to perform rapid and stable measurements is even more pronounced.Therefore, the technique of the present invention can be used more effectively. In this case, the problem can be solved more effectively by removing the sol after the solvent.

[0093] The cell immobilization kit of the present disclosure comprises a gel component that can be converted into a sol, a reagent, and / or an instrument (e.g., a cell culture vessel) used for immobilizing cells with a gel. The gel component that can be converted into a sol in the kit may be in powder, gel, or sol form. The cell culture vessel in the kit has compartments for immobilizing cells. The base area of ​​a single compartment is not particularly limited. From the perspective of detection sensitivity or manufacturing efficiency, the base area is preferably 0.5 to 100 mm². 2 , even more preferably 1 to 30 mm 2 and even more preferably 1.5 to 15 mm2 In one embodiment of the present invention, the upper limit of the area can be set to 80 mm. 2 , 60 mm 2 or 40 mm 2 be determined.

[0094] As described above, transportable cells that can be stably transported can be produced by including the step of immobilizing cells with a gel that can be converted into a sol. After transport, the gel immobilizing the transportable cells can be converted into a sol and removed upon use. This allows the use of cells without gel or sol after stable transport, thus enabling effective light detection from cells, which can be problematic, for example, with a highly turbid gel or sol. Therefore, in one embodiment, the present disclosure relates to a method for producing a transportable cell that includes immobilizing a cell with a gel that can be converted into a sol.Furthermore, in one embodiment, the present disclosure relates to a method for detecting light from a cell, comprising the conversion of a gel into a sol, wherein the gel immobilizes the cell and can be converted into the sol. In the light detection method, a treatment can optionally be carried out after the solation to cause the cells to emit light (e.g., addition of a substance or irradiation with light). EXAMPLES

[0095] The present invention is described in detail below with reference to examples. However, the present invention is not limited to these examples. Test example 1: Production of a stably expressing cell

[0096] Cells derived from Spilosoma imparilis (SpIm cells) were transfected with a transposon vector containing the coding sequence for an olfactory receptor protein (ORA), the coding sequence for an olfactory receptor co-receptor, the coding sequence for a calcium sensor fluorescence protein, and the coding sequence for a puromycin resistance gene, all under the control of a promoter sequence and positioned between the 5' inverted terminal repeat (ITW) and the 3' ITW. Selection was performed using puromycin, resulting in SpIm cells that stably express olfactory receptors in which the foreign DNA containing the aforementioned coding sequences and the promoter sequence was integrated into the chromosomal genomic DNA (hereinafter referred to as "ORA cells"). The olfactory receptor is an insect-derived olfactory receptor and a receptor for compound a. The ORA cells emit fluorescence in response to compound a. Test example 2. Preparation of a gelatin solution

[0097] Powdered fish gelatin (fish gelatin (Type A) manufactured by Nitta Gelatin Inc.) was added to a PBS solution, and the mixture was stirred at 17 °C for 30 minutes. The mixture was then heated to 35 °C and stirred until completely dissolved, yielding a gelatin solution with a predetermined fish gelatin concentration. The solution was sterilized using a 0.22 µm filter or by autoclaving before use in the following tests. Test example 3: Production of an acrylic resin solution

[0098] 2-[[2-(Methacryloyloxy)ethyl]dimethylammonio]acetate (monomer: MO3N6O) and N,N'-[Oxybis(2,1-ethanediyl-3,1-propanediyl)]bisacrylamide (crosslinking agent: 2AAmLN) were dissolved in an aqueous PBS solution (Nissui Pharmaceutical Co., Ltd. #05913) to prepare an aqueous MO3N6O / 2AAmLN solution. Lithium phenyl(2,4,6-trimethylbenzoyl)phosphinate (photopolymerization initiator: LPA) was dissolved in an aqueous PBS solution to prepare an aqueous LPA solution. The two aqueous solutions mentioned above were mixed and stirred so that the final concentration of MO3N6O was 0.8 wt%, the final concentration of 2AAmLN was 5.2 wt% and the final concentration of LPA was 0.02 wt%, resulting in an acrylic resin solution. Test example 4. Measurement of olfactory receptor activity 1

[0099] ORA cells were cultured in a Corning 3903 96-well plate with 5 × 10 4Cells / 100 µL / well were seeded and incubated at 27 °C (without CO2 injection). Sf-900 III SFM medium was used as the culture medium. The following procedures were performed 24 hours after seeding.

[0100] Test group 1: The culture fluid was removed from each well of the 96-well plate, and 40 µL of the acrylic resin solution was added to each well. The cells were irradiated with light at a wavelength of 365 nm at 110 mW / cm². 2 The sample was irradiated for 30 seconds using a UV irradiation device (LED light source, 365 nm, manufactured by CCS Inc.) to form a hydrogel. 210 µL of culture medium were added to the hydrogel, followed by two washes with culture medium.

[0101] Test group 2: Cultivation continued without gel formation.

[0102] Twenty-four hours after the above procedures, the medium was completely removed, and 40 µl of 0.1% BSA / 1x Hanks buffer / 20 mM HEPES buffer was added to each well of test group 1, while 80 µl of the same buffer was added to each well of test group 2. Compound a (a substance to which ORA cells respond) was added to each well to achieve a final concentration of 100 µM, and the changes in fluorescence intensity before and after the addition of compound a were quantified using a microplate reader (FlexStation 3, Molecular Devices).

[0103] Fig.Figure 1 shows the results. In the absence of a hydrogel (test group 2), the fluorescence intensity increased sharply after the addition of compound a, and the fluorescence intensity was measured quickly and stably. In the presence of a hydrogel (test group 1), however, the fluorescence intensity increased gradually after the addition of compound a, and a quick and stable measurement of the fluorescence intensity was not possible. Test example 5: Measurement of olfactory receptor activity 2

[0104] ORA cells were cultured in a Corning 3903 96-well plate with 5 × 10 4 Seeds were sown at 100 µL / well and incubated at 27 °C (without CO2 injection). Sf-900 III SFM medium was used as the culture medium. The following procedures were carried out 24 hours after sowing. Test group A: The culture fluid was removed from each well of the 96-well plate, and 40 µL of a gelatin solution (sol with a fish gelatin concentration of 5 wt%) was added to each well at 25 °C. Gel formation was achieved by standing for 3 hours at 4 °C. Test group B: Same as test group A. Test group C: Cultivation continued without gel formation.

[0105] Following the steps described above, test groups A and B were left to stand at 27 °C for 1 hour to convert the gels into sols. In test group A, 40 µl of 0.1% BSA / 1x Hanks buffer / 20 mM HEPES buffer was added to each well without removing the sol. In test groups B and C, the sol or culture medium was removed, and then 80 µl of 0.1% BSA / 1x Hanks buffer / 20 mM HEPES buffer was added to each well. Compound a (a substance to which ORA cells respond) was added to each well to achieve a final concentration of 100 µM, and the changes in fluorescence intensity before and after the addition of compound a were quantified using a microplate reader (FlexStation 3, Molecular Devices).

[0106] Fig.Figure 2 shows the results. In the case where the gel was converted into a sol (test group A), the fluorescence intensity increased sharply after the addition of compound a, enabling a rapid and stable measurement of the fluorescence intensity. A comparison of test group A with test groups B and C showed that immobilizing the cells with a gel had only a minor effect on the speed and stability of the chemical detection. Test example 6: Measurement of olfactory receptor activity 3

[0107] ORA cells were cultured in a Corning 3903 96-well plate with 5 × 10 4 Cells were seeded at 100 µL / well and incubated at 27 °C (without CO2 injection). Sf-900 III SFM medium was used as the culture medium.

[0108] Twenty-four hours after seeding, the culture fluid was removed from each well of the 96-well plate, and 40 µL of a gelatin solution at 25 °C (sol with a fish gelatin concentration of 2 to 8 wt%) was added to each well. Gel formation was allowed by incubating for at least 1 hour at 4 °C. 100 µL of culture medium was then added to the gel. An additional group was prepared in which cultivation continued with 100 µL of culture medium without replacing the medium with the gelatin solution. After gel formation, the samples were stored at 4 °C.

[0109] Seventy-two hours after gel formation, the gel was incubated at 27 °C for 1 hour to convert it to a sol. After sol removal, 80 µl of 0.1% BSA / 1x Hanks buffer / 20 mM HEPES buffer was added to each well. Subsequently, each well was treated with compound a (a substance to which ORA cells respond) to achieve final concentrations of 0, 0.1, 1, and 10 µM, and the changes in fluorescence intensity before and after the addition of compound a were quantified using a microplate reader (FlexStation 3, Molecular Devices).

[0110] Fig. Figure 3 shows the results. The chemical substance was detected in all cases where the gel was used in different concentrations. The chemical substance was also detected when the cells were immobilized with a hydrogel and stored at 4 °C. Test example 7: Measurement of olfactory receptor activity 4

[0111] ORA cells were cultured in a Corning 3903 96-well plate with 5 × 10 4 Cells were seeded at 100 µl / well and incubated at 27 °C (without CO2 injection). Sf-900 III SFM medium was used as the culture medium.

[0112] Twenty-four hours after sowing, the culture fluid was removed from each well of the 96-well plate, and 40 µL of a gelatin solution (sol with a fish gelatin concentration of 5 wt%) was added to each well at 25 °C. Gel formation was allowed by incubating for 3 hours at 4 °C. An additional group was prepared in which cultivation continued with 100 µL of culture medium without replacing the medium with the gelatin solution.

[0113] After gel formation, the gel was immediately incubated at 27 °C for 1 hour to convert it to a sol. After sol removal, 80 µl of 0.1% BSA / 1x Hanks buffer / 20 mM HEPES buffer was added to each well. Alternatively, after solation, 40 µl of 0.1% BSA / 1x Hanks buffer / 20 mM HEPES buffer was added without removing the sol. Compound a (a substance to which ORA cells respond) was added to each well to achieve final concentrations of 0, 0.1, 1, and 10 µM, and the changes in fluorescence intensity before and after the addition of compound a were quantified using a microplate reader (FlexStation 3, Molecular Devices).

[0114] Fig. Figure 4 shows the results. The chemical substance was detected regardless of whether the sol was removed or not. However, the detection sensitivity was higher when the sol was removed. Test example 8: Measurement of solvency 1

[0115] A gelatin solution (sol with a fish gelatin concentration of 2 to 5 wt%) was placed in a sample tube and incubated in a column oven at 40 °C for 1 hour to eliminate any thermal history. The sample tube was then transferred to an incubator at 25 °C and left to stand for 30 minutes. The sample tube was then refrigerated at 4 °C for 24 hours to allow gelation. Finally, the sample tube was transferred to an incubator at 25 °C and left to stand for 1 hour to allow solation. Viscosity was measured using a Kyoto Electronics Manufacturing Co., Ltd. EMS-1000 viscometer with a 2 mm aluminum probe at a measurement temperature of 25 °C.

[0116] Fig. Figure 5 shows the results. The gel formed from the gelatin solution (sol with a fish gelatin concentration of 2 to 5%) formed a sol with a relatively low viscosity after solation. Test example 9: Sol viscosity measurement 2

[0117] A gelatin solution (sol with a fish gelatin concentration of 2 to 5 wt%) was placed in a sample tube and incubated in a column oven at 40 °C for 1 hour to eliminate any thermal history. The sample tube was then placed in an incubator at 25 °C and left to stand for 30 minutes. The sample tube was then refrigerated at 4 °C for 2 days to allow gelation. Finally, the sample tube was transferred to an incubator at 25 °C and left to stand for 2 days to allow solation. Viscosity was measured using a Kyoto Electronics Manufacturing Co., Ltd. EMS-1000 viscometer with a 2 mm aluminum probe at a measurement temperature of 25 °C.

[0118] Fig. Figure 6 shows the results. A comparison between Fig.5, which shows the measurement results one hour after the separation, and Fig. Figure 6, which shows the measurement results two days after the solvent, indicates that the viscosity of the solvent increases over time at relatively high concentrations after solventing. Test example 10: Measurement of the storage circuit module of gel and the sol-gel transition temperature

[0119] The measurements were performed using an Anton Paar Japan KK MCR302 viscoelasticity meter, a CP50-1 plate with a sample volume of 1 ml, a frequency of 1 Hz, and a strain of 1%. Viscosity measurements were initially taken at 25 °C, followed by temperature measurements by cooling to 4 °C at a rate of -1 °C / min. The sol-gel transition point was defined as the point at which the loss factor falls below 1. Subsequently, the storage elastic modulus was measured 10 minutes after reaching 4 °C.

[0120] Fig.Figure 7 and Table 1 show the results. The storage elasticity modulus at 4 °C was 1.3 kPa for 5 wt.%, 1.1 kPa for 4 wt.%, 0.3 kPa for 3 wt.%, 0.1 kPa for 2 wt.% and 0.002 kPa for 1 wt.%. Table 1 Gelatin concentration (wt%) Sol-gel transition temperature (°C) 10 17 5 13 4 12 3 9 2 5 QUOTES INCLUDED IN THE DESCRIPTION

[0000] This list of documents cited by the applicant was automatically generated and is included solely for the reader's convenience. The list is not part of the German patent or utility model application. The DPMA accepts no liability for any errors or omissions. Cited patent literature

[0000] WO 2022 / 024902A

[0004] Cited non-patent literature

[0000] K. Hofmann, W. Stoffel, TMbase - eine Datenbank mit Segmenten von Membran-spannenden Proteinen, Biol. Chem. Hoppe-Seyler, 374 (1993), S. 166, https: / / embnet.vital-it.ch / software / TMPRED_form.html

[0046] Karlin S., Altschul S. F. Methods for assessing the statistical significance of molecular sequence features by using general scorings schemes, Proc. Natl. Acad. Sci. USA. 87: 2264-2268 (1990

[0054] Karlin S., Altschul S. F. Applications and statistics for multiple high-scoring segments in molecular sequences, Proc Natl Acad Sci USA. 90: 5873-7 (1993

[0054]

Claims

[1] Cell chip comprising a compartment containing a cell and a gel that can be converted into a sol. [2] Cell chip according to claim 1, wherein the gel is a gel which can be converted into a sol by temperature control, light irradiation or the addition of a chemical agent. [3] Cell chip according to claim 2, wherein the gel is a gel which can be converted into a sol by temperature control. [4] Cell chip according to claim 3, wherein the gel has a sol formation temperature of 40 °C or less. [5] Cell chip according to claim 3, wherein the gel has a sol formation temperature of 30 °C or less. [6] Cell chip according to claim 3, wherein the gel has a sol-gel transition temperature of 4 to 25 °C. [7] Cell chip according to claim 1, wherein the sol formed after solation of the gel has a viscosity of 20 mPa·s or less. [8] Cell chip according to claim 1, wherein the gel has a storage elasticity modulus of 0.01 to 20 kPa at 4 °C. [9] Cell chip according to at least one of claims 1 to 8 for use in a method for detecting a chemical test substance, wherein the method comprises bringing the cell into contact with the chemical test substance after the solventing of the gel. [10] Cell chip according to at least one of claims 1 to 8 for use in the detection of light from the cell. [11] Cell chip according to at least one of claims 1 to 8, wherein the cell is an insect cell. [12] Cell chip according to at least one of claims 1 to 8, wherein the cell comprises an exogenous polynucleotide containing a coding sequence for a sensor protein. [13] Cell chip according to claim 12, wherein the sensor protein is an odor receptor protein. [14] Kit for immobilizing a cell, comprising a component for forming a gel which can be converted into a sol. [15] Kit according to claim 14 for immobilizing a cell on a cell chip. [16] Method for producing a cell for transport, comprising immobilizing a cell with a gel that can be converted into a sol. [17] Method for detecting light from a cell, comprising converting a gel into a sol, wherein the gel can immobilize the cell and be converted into a sol. [18] Gel that can be converted into a sol, where the gel can be converted into a sol by temperature control the gel has a sol formation temperature of 40 °C or less, the gel has a sol-gel transition temperature of 4 to 25 °C, the sol formed after solation of the gel has a viscosity of 20 mPa·s or less and The gel has a storage elasticity modulus of 0.01 to 20 kPa at 4 °C. [19] Sol for forming the gel according to claim 18. [20] Kit according to claim 14, comprising the gel according to claim 18 and / or the sol according to claim 19. [21] Kit according to claim 20 for immobilizing a cell on a cell chip. [22] Method according to claim 16, wherein the gel is the gel according to claim 18. [23] Method according to claim 17, wherein the gel is the gel according to claim 18.

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