JP2020526767A - ヒト混合細胞集団スフェロイドの高スループット光学アッセイ - Google Patents
ヒト混合細胞集団スフェロイドの高スループット光学アッセイ Download PDFInfo
- Publication number
- JP2020526767A JP2020526767A JP2020501302A JP2020501302A JP2020526767A JP 2020526767 A JP2020526767 A JP 2020526767A JP 2020501302 A JP2020501302 A JP 2020501302A JP 2020501302 A JP2020501302 A JP 2020501302A JP 2020526767 A JP2020526767 A JP 2020526767A
- Authority
- JP
- Japan
- Prior art keywords
- cells
- spheroids
- well
- fluorescence
- plate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000003287 optical effect Effects 0.000 title claims description 13
- 238000003556 assay Methods 0.000 title description 20
- 238000000034 method Methods 0.000 claims abstract description 33
- 210000005260 human cell Anatomy 0.000 claims abstract description 14
- 210000004027 cell Anatomy 0.000 claims description 54
- 210000002569 neuron Anatomy 0.000 claims description 26
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 21
- 229910052791 calcium Inorganic materials 0.000 claims description 21
- 239000011575 calcium Substances 0.000 claims description 21
- 210000000130 stem cell Anatomy 0.000 claims description 16
- 210000002889 endothelial cell Anatomy 0.000 claims description 15
- 150000001875 compounds Chemical class 0.000 claims description 14
- 210000001130 astrocyte Anatomy 0.000 claims description 12
- 230000008859 change Effects 0.000 claims description 11
- 210000002919 epithelial cell Anatomy 0.000 claims description 11
- 210000004248 oligodendroglia Anatomy 0.000 claims description 9
- 238000012360 testing method Methods 0.000 claims description 9
- 210000002216 heart Anatomy 0.000 claims description 7
- 210000003734 kidney Anatomy 0.000 claims description 7
- 210000004185 liver Anatomy 0.000 claims description 7
- 230000002025 microglial effect Effects 0.000 claims description 7
- 210000000496 pancreas Anatomy 0.000 claims description 7
- 210000004072 lung Anatomy 0.000 claims description 6
- 210000005265 lung cell Anatomy 0.000 claims description 6
- 206010028980 Neoplasm Diseases 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 5
- 210000003038 endothelium Anatomy 0.000 claims description 5
- 210000002064 heart cell Anatomy 0.000 claims description 5
- 210000003292 kidney cell Anatomy 0.000 claims description 5
- 210000005229 liver cell Anatomy 0.000 claims description 5
- 201000011510 cancer Diseases 0.000 claims description 4
- 210000003668 pericyte Anatomy 0.000 claims description 4
- 210000000170 cell membrane Anatomy 0.000 claims description 3
- 238000000799 fluorescence microscopy Methods 0.000 abstract description 3
- 238000002825 functional assay Methods 0.000 abstract description 2
- 238000010586 diagram Methods 0.000 abstract 1
- 229940079593 drug Drugs 0.000 description 12
- 239000003814 drug Substances 0.000 description 12
- 230000001537 neural effect Effects 0.000 description 11
- 239000005557 antagonist Substances 0.000 description 6
- 230000010355 oscillation Effects 0.000 description 5
- 230000004044 response Effects 0.000 description 5
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- 239000000556 agonist Substances 0.000 description 4
- 230000003371 gabaergic effect Effects 0.000 description 4
- 230000000848 glutamatergic effect Effects 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- IYGYMKDQCDOMRE-QRWMCTBCSA-N Bicculine Chemical compound O([C@H]1C2C3=CC=4OCOC=4C=C3CCN2C)C(=O)C2=C1C=CC1=C2OCO1 IYGYMKDQCDOMRE-QRWMCTBCSA-N 0.000 description 2
- 206010010904 Convulsion Diseases 0.000 description 2
- 206010013710 Drug interaction Diseases 0.000 description 2
- 102000034615 Glial cell line-derived neurotrophic factor Human genes 0.000 description 2
- 108091010837 Glial cell line-derived neurotrophic factor Proteins 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- AACMFFIUYXGCOC-UHFFFAOYSA-N bicuculline Natural products CN1CCc2cc3OCOc3cc2C1C4OCc5c6OCOc6ccc45 AACMFFIUYXGCOC-UHFFFAOYSA-N 0.000 description 2
- 230000003185 calcium uptake Effects 0.000 description 2
- IYGYMKDQCDOMRE-UHFFFAOYSA-N d-Bicucullin Natural products CN1CCC2=CC=3OCOC=3C=C2C1C1OC(=O)C2=C1C=CC1=C2OCO1 IYGYMKDQCDOMRE-UHFFFAOYSA-N 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 125000001153 fluoro group Chemical group F* 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 210000000274 microglia Anatomy 0.000 description 2
- 230000003278 mimic effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 210000000225 synapse Anatomy 0.000 description 2
- 230000000946 synaptic effect Effects 0.000 description 2
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 description 1
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 238000000423 cell based assay Methods 0.000 description 1
- 239000013553 cell monolayer Substances 0.000 description 1
- 210000002236 cellular spheroid Anatomy 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- BFMYDTVEBKDAKJ-UHFFFAOYSA-L disodium;(2',7'-dibromo-3',6'-dioxido-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl)mercury;hydrate Chemical compound O.[Na+].[Na+].O1C(=O)C2=CC=CC=C2C21C1=CC(Br)=C([O-])C([Hg])=C1OC1=C2C=C(Br)C([O-])=C1 BFMYDTVEBKDAKJ-UHFFFAOYSA-L 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 208000028329 epileptic seizure Diseases 0.000 description 1
- 230000000763 evoking effect Effects 0.000 description 1
- 238000012921 fluorescence analysis Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000013537 high throughput screening Methods 0.000 description 1
- 238000012188 high-throughput screening assay Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/682—Signal amplification
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/067—Hepatocytes
- C12N5/0671—Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5058—Neurological cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
- G01N33/6851—Methods of protein analysis involving laser desorption ionisation mass spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/251—Colorimeters; Construction thereof
- G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Computational Biology (AREA)
- Organic Chemistry (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Genetics & Genomics (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Optics & Photonics (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Gastroenterology & Hepatology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
【選択図】図4
Description
この出願は、2017年7月14日に出願された米国出願第62/532,667号の出願日の利益を主張し、その開示は参照により本明細書に組み込まれる。
創薬のペースの加速が、生きたヒト細胞を使用した機能的なin vitroアッセイの必要性の増加を引き起こしている。しかしながら、ハイスループットシステム用にこれらのアッセイを自動化することは困難であることが判明している。ハイスループット設定で使用される最も一般的な生物学的アッセイは、ある種の蛍光測定に依存していることに留意されたい。
驚くべきことに、3Dスフェロイド上の薬物分子によるカルシウム振動の変調を監視することができ、結果のデータは非常に一貫性がある。たとえば、スフェロイド神経細胞シナプスは長期間にわたって予測可能かつ一貫して興奮する。一実施形態では、本開示は、3Dヒト細胞スフェロイド、例えば混合集団ヒト細胞ニューロンスフェロイドの光学アッセイ、例えば機能的FLIPRアッセイ又は高含有量高倍率光学顕微鏡法を提供する。一実施形態では、試験前に、スフェロイドを4〜16週間培養して、成熟したヒト様脳機能を模倣するために、堅牢な同期シナプスネットワークを誘導する。これらの混合集団のスフェロイドは、長期間にわたって予測可能かつ一貫して興奮する。一実施形態では、本開示は、FLIPR及びカルシウム取り込み蛍光振動(calcium uptake fluorescence oscillations)を利用する、ヒト細胞3Dスフェロイドの混合集団のハイスループット光学アッセイを提供する。振動は化学化合物で調整でき、振動性の興奮はアゴニスト又はアンタゴニストで変更可能である。
以下の議論は、本発明の様々な実施形態に向けられている。これらの実施形態の1つ以上が好ましい場合があるが、本発明は開示された実施形態に限定されない。加えて、当業者は、以下の説明が広範な用途を有し、任意の実施形態の議論はその実施形態の例示に過ぎず、開示又は特許請求の範囲をその実施形態に限定することを意図しないことを理解するであろう。
スフェロイド、例えば2つ以上の異なる細胞タイプから形成されたものは、任意に1又は複数の異なる成長因子を含む任意の適切な培地、及び任意の適切な条件を使用して調製され得る。例えば、神経細胞及びアストロサイトから形成されたスフェロイドは、一実施形態では、1又は複数の以下の培地及び/又は条件を使用して調製されてもよい:SM1 Neuronal Supplement(BrainPhys(商標) Neuronal Medium及びSM1 Kit(Cat.#05792; StemCell Technologies))、20ng/mL BDNF(cat.#78005; StemCell Technologies)、20ng/mL GDNF(cat.#78058; StemCell Technologies)及びペニシリン/ストレプトマイシン(cat.#SV30010; GE Healthcare Life Sciences)で1倍補足されたBrainPhys(商標)Neuronal Medium(StemCell Tech)。細胞は、5%CO2及び高湿度のインキュベーター内で37℃に維持される。
Claims (28)
- スフェロイドに対する1又は複数の化合物の効果を検出する光学的方法であって、
均一な直径のヒト細胞の1又は複数のスフェロイドと、1又は複数の試験化合物とを接触させ;そして
前記スフェロイドの1又は複数の振動の量又は変化を光学的に検出すること、
を含む、方法。 - 前記1又は複数のスフェロイドがマルチウェルプレートのウェル内にある、請求項1に記載の方法。
- 各ウェルが1つのスフェロイドを有する、請求項2に記載の方法。
- 前記ウェルが、カルシウムを検出するのに有用な蛍光分子とさらに接触され、経時的な蛍光の量又は変化が、1又は複数のウェルで検出される、請求項2又は3に記載の方法。
- 前記蛍光の量又は変化が、蛍光のピークの量、1又は複数のピークの振幅、1又は複数のピークの間のピーク間隔、1又は複数のピークの幅、あるいはそれらの任意の組み合わせを検出することである、請求項4に記載の方法。
- 前記1又は複数のスフェロイドが、神経細胞を含む、請求項1〜5のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、神経細胞及びアストロサイトを含む、請求項1〜5のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、心臓、肝臓、腎臓、膵臓、肺、内皮又は上皮細胞を含む、請求項1〜7のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、癌細胞又は不死化細胞を含む、請求項1〜8のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、ミクログリア細胞又はオリゴデンドロサイトを含む、請求項1〜9のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、周皮細胞及び内皮細胞を含む、請求項1〜10のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、内皮細胞、ミクログリア細胞、神経細胞、オリゴデンドロサイト細胞、又はそれらの任意の組み合わせを含む、請求項1〜11のいずれか一項に記載の方法。
- 前記細胞が、前駆細胞である、請求項1〜12のいずれか一項に記載の方法。
- 前記前駆細胞が、神経細胞、アストロサイト、心臓細胞、肝臓細胞、腎臓細胞、膵臓細胞、肺細胞、内皮細胞、又は上皮細胞の前駆細胞である、請求項13に記載の方法。
- 前記1又は複数のスフェロイドが、約500〜約600ミクロンの直径を有する、請求項1〜14のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、約450〜約500ミクロンの直径を有する、請求項1〜14のいずれか一項に記載の方法。
- 前記1又は複数のスフェロイドが、前記1又は複数の試験化合物と接触する前に少なくとも4〜6週間培養される、請求項1〜16のいずれか一項に記載の方法。
- 前記蛍光分子が、カルシウム3、カルシウム4、カルシウム5、カルシウム6、Fluo3、又はFluo4を含む、請求項3〜17のいずれか一項に記載の方法。
- 前記ウェルを、細胞膜非透過性クエンチャーと接触させることをさらに含む、請求項3〜18のいずれか一項に記載の方法。
- 前記蛍光の変化の量が、スフェロイド及び蛍光分子を含むが試験化合物を含まないウェル内の蛍光と比較される、請求項3〜19のいずれか一項に記載の方法。
- ウェルあたり1又は複数の混合ヒト細胞スフェロイドを含むマルチウェルプレート。
- 前記1又は複数のスフェロイドが、神経細胞及びアストロサイトを含む、請求項21に記載のプレート。
- 前記1又は複数のスフェロイドが、心臓、肝臓、腎臓、膵臓、肺、内皮又は上皮細胞を含む、請求項21又は22に記載のプレート。
- 前記1又は複数のスフェロイドが、ミクログリア細胞又はオリゴデンドロサイトを含む、請求項21〜23のいずれか一項に記載のプレート。
- 前記1又は複数のスフェロイドが、周皮細胞及び内皮細胞を含む、請求項21〜24のいずれか一項に記載のプレート。
- 前記1又は複数のスフェロイドが、内皮細胞、ミクログリア細胞、神経細胞、オリゴデンドロサイト細胞、又はそれらの任意の組み合わせを含む、請求項21〜25のいずれか一項に記載のプレート。
- 前記1又は複数のスフェロイドが、神経細胞、アストロサイト、心臓細胞、肝臓細胞、腎臓細胞、膵臓細胞、肺細胞、内皮細胞、又は上皮細胞の前駆細胞を含む、請求項21〜26のいずれか一項に記載のプレート。
- 前記ウェルが、異なる細胞から形成されたスフェロイドを含む、請求項21〜27のいずれか一項記載のプレート。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2022019584A JP2022065065A (ja) | 2017-07-14 | 2022-02-10 | ヒト混合細胞集団スフェロイドの高スループット光学アッセイ |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201762532667P | 2017-07-14 | 2017-07-14 | |
US62/532,667 | 2017-07-14 | ||
PCT/US2018/042105 WO2019014603A1 (en) | 2017-07-14 | 2018-07-13 | HIGH PERFORMANCE OPTICAL DETERMINATION OF POPULATION SPHEREOIDS OF HUMAN MIXED CELLS |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2022019584A Division JP2022065065A (ja) | 2017-07-14 | 2022-02-10 | ヒト混合細胞集団スフェロイドの高スループット光学アッセイ |
Publications (1)
Publication Number | Publication Date |
---|---|
JP2020526767A true JP2020526767A (ja) | 2020-08-31 |
Family
ID=63174393
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2020501302A Pending JP2020526767A (ja) | 2017-07-14 | 2018-07-13 | ヒト混合細胞集団スフェロイドの高スループット光学アッセイ |
JP2022019584A Pending JP2022065065A (ja) | 2017-07-14 | 2022-02-10 | ヒト混合細胞集団スフェロイドの高スループット光学アッセイ |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2022019584A Pending JP2022065065A (ja) | 2017-07-14 | 2022-02-10 | ヒト混合細胞集団スフェロイドの高スループット光学アッセイ |
Country Status (4)
Country | Link |
---|---|
US (1) | US11193159B2 (ja) |
EP (1) | EP3652536A1 (ja) |
JP (2) | JP2020526767A (ja) |
WO (1) | WO2019014603A1 (ja) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3918056A1 (en) * | 2019-02-02 | 2021-12-08 | Stemonix Inc. | Method of using human spheroids for drug discovery |
US20210371784A1 (en) | 2020-05-27 | 2021-12-02 | Ricoh Company, Ltd. | Cell-containing vessel and method for producing neural cell-containing spheroid |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014141528A1 (ja) * | 2013-03-15 | 2014-09-18 | 国立大学法人佐賀大学 | 心臓又は血管組織型スフェロイド |
WO2016115489A1 (en) * | 2015-01-16 | 2016-07-21 | Clemson University Research Foundation | Tissues containing semiconductor nanomaterials and methods of preparing and using the same |
US20170115275A1 (en) * | 2015-10-23 | 2017-04-27 | Arizona Board Of Regents On Behalf Of Arizona State University | Engineered substrates for high-throughput generation of 3d models of tumor dormancy, relapse and micrometastases for phenotype specific drug discovery and development |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6214563B1 (en) * | 1997-08-01 | 2001-04-10 | Aurora Biosciences Corporation | Photon reducing agents for reducing undesired light emission in assays |
US7615356B2 (en) * | 2000-07-10 | 2009-11-10 | Vertex Pharmaceuticals (San Diego) Llc | Ion channel assay methods |
CA2772945A1 (en) * | 2009-09-25 | 2011-03-31 | Xoma Technology Ltd. | Screening methods |
US8501476B2 (en) * | 2009-10-07 | 2013-08-06 | Brown University | Assays and methods for fusing cell aggregates to form proto-tissues |
US20110143960A1 (en) * | 2009-12-10 | 2011-06-16 | Labarbera Daniel V | 3d-models for high-throughput screening drug discovery and development |
US9334473B2 (en) * | 2012-08-17 | 2016-05-10 | Jelena Vukasinovic | Three dimensional cell culture compositions and methods of use |
-
2018
- 2018-07-13 US US16/035,039 patent/US11193159B2/en active Active
- 2018-07-13 EP EP18753497.9A patent/EP3652536A1/en active Pending
- 2018-07-13 WO PCT/US2018/042105 patent/WO2019014603A1/en unknown
- 2018-07-13 JP JP2020501302A patent/JP2020526767A/ja active Pending
-
2022
- 2022-02-10 JP JP2022019584A patent/JP2022065065A/ja active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014141528A1 (ja) * | 2013-03-15 | 2014-09-18 | 国立大学法人佐賀大学 | 心臓又は血管組織型スフェロイド |
WO2016115489A1 (en) * | 2015-01-16 | 2016-07-21 | Clemson University Research Foundation | Tissues containing semiconductor nanomaterials and methods of preparing and using the same |
US20170115275A1 (en) * | 2015-10-23 | 2017-04-27 | Arizona Board Of Regents On Behalf Of Arizona State University | Engineered substrates for high-throughput generation of 3d models of tumor dormancy, relapse and micrometastases for phenotype specific drug discovery and development |
Non-Patent Citations (2)
Title |
---|
STEPHANIE M. RAVENSCROFT ET AL: "Cardiac Non-myocyte Cells Show Enhanced Pharmacological Function Suggestive of Contractile Maturity", TOXICOLOGICAL SCIENCES, vol. 152, no. 1, JPN7021000642, 28 April 2016 (2016-04-28), pages 99 - 112, XP055504282, ISSN: 0004615182, DOI: 10.1093/toxsci/kfw069 * |
TERRASSO ANA PAULA ET AL: "Novel scalable 3D cell based model for in vitro neurotoxicity testing: Combining human differentiate", JOURNAL OF BIOTECHNOLOGY, vol. 205, JPN6021007164, 5 January 2015 (2015-01-05), pages 82 - 92, ISSN: 0004615183 * |
Also Published As
Publication number | Publication date |
---|---|
US20190017097A1 (en) | 2019-01-17 |
US11193159B2 (en) | 2021-12-07 |
EP3652536A1 (en) | 2020-05-20 |
JP2022065065A (ja) | 2022-04-26 |
WO2019014603A1 (en) | 2019-01-17 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Renner et al. | A fully automated high-throughput workflow for 3D-based chemical screening in human midbrain organoids | |
Di Rienzo et al. | Probing short-range protein Brownian motion in the cytoplasm of living cells | |
Robinette et al. | In vitro assessment of developmental neurotoxicity: use of microelectrode arrays to measure functional changes in neuronal network ontogeny | |
Vergara et al. | Three-dimensional automated reporter quantification (3D-ARQ) technology enables quantitative screening in retinal organoids | |
JP7034354B2 (ja) | 生存細胞の画像解析のための解析方法 | |
Jonsson et al. | Impedance-based detection of beating rhythm and proarrhythmic effects of compounds on stem cell-derived cardiomyocytes | |
JP2022065065A (ja) | ヒト混合細胞集団スフェロイドの高スループット光学アッセイ | |
EP2157166B1 (en) | Model cell chip, apparatus for evaluating drug effect using the model cell chip and method of evaluating drug effect | |
Charwat et al. | Potential and limitations of microscopy and Raman spectroscopy for live-cell analysis of 3D cell cultures | |
Hansen et al. | Optical method to quantify mechanical contraction and calcium transients of human pluripotent stem cell-derived cardiomyocytes | |
Pesl et al. | Phenotypic assays for analyses of pluripotent stem cell–derived cardiomyocytes | |
Robinson et al. | A novel toolkit for characterizing the mechanical and electrical properties of engineered neural tissues | |
Son et al. | Electrophysiological monitoring of neurochemical-based neural signal transmission in a human brain–spinal cord assembloid | |
WO2019031545A1 (ja) | 多能性幹細胞の未分化状態を判定する方法、多能性幹細胞の継代培養方法およびそれら方法に使用される装置 | |
Lee et al. | Repeated and on-demand intracellular recordings of cardiomyocytes derived from human-induced pluripotent stem cells | |
Glaser et al. | Intracellular calcium measurements for functional characterization of neuronal phenotypes | |
Tukker et al. | In vitro techniques for assessing neurotoxicity using human iPSC-derived neuronal models | |
Pfeiffer et al. | Optimized temporally deconvolved Ca2+ imaging allows identification of spatiotemporal activity patterns of CA1 hippocampal ensembles | |
Chirieleison et al. | Automated live cell imaging systems reveal dynamic cell behavior | |
Boddum et al. | Optogenetics and optical tools in automated patch clamping | |
JP2024501849A (ja) | 光学プレートコントローラおよびリーダ | |
Sevetson et al. | Cortical spheroids display oscillatory network dynamics | |
Frega | Neuronal network dynamics in 2D and 3D in vitro neuroengineered systems | |
Forny et al. | Contractions of Human-iPSC-derived Cardiomyocyte Syncytia Measured with a Ca-sensitive Fluorescent Dye in Temperature-controlled 384-well Plates | |
George et al. | Decoding Ca 2+ Signals as a Non-electrophysiological Method for Assessing Drug Toxicity in Stem Cell-Derived Cardiomyocytes |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20200303 |
|
A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20210224 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210302 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210602 |
|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20211012 |