WO2009029039A1 - Procédé de mesure de la viabilité d'une cellule - Google Patents
Procédé de mesure de la viabilité d'une cellule Download PDFInfo
- Publication number
- WO2009029039A1 WO2009029039A1 PCT/SE2008/050974 SE2008050974W WO2009029039A1 WO 2009029039 A1 WO2009029039 A1 WO 2009029039A1 SE 2008050974 W SE2008050974 W SE 2008050974W WO 2009029039 A1 WO2009029039 A1 WO 2009029039A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- measurement
- status
- marker
- viability
- Prior art date
Links
- 238000005259 measurement Methods 0.000 title claims abstract description 47
- 238000000034 method Methods 0.000 title claims abstract description 43
- 230000003833 cell viability Effects 0.000 title description 11
- 239000003550 marker Substances 0.000 claims abstract description 30
- 230000035899 viability Effects 0.000 claims abstract description 18
- 239000007787 solid Substances 0.000 claims abstract description 12
- 230000002285 radioactive effect Effects 0.000 claims abstract description 9
- 230000005855 radiation Effects 0.000 claims abstract description 4
- 229910052804 chromium Inorganic materials 0.000 claims description 4
- 239000011651 chromium Substances 0.000 claims description 4
- WPUZGNPQMIWOHE-UHFFFAOYSA-N 3',6'-diacetyloxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-5-carboxylic acid Chemical compound O1C(=O)C2=CC(C(O)=O)=CC=C2C21C1=CC=C(OC(C)=O)C=C1OC1=CC(OC(=O)C)=CC=C21 WPUZGNPQMIWOHE-UHFFFAOYSA-N 0.000 claims description 2
- NJYVEMPWNAYQQN-UHFFFAOYSA-N 5-carboxyfluorescein Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C21OC(=O)C1=CC(C(=O)O)=CC=C21 NJYVEMPWNAYQQN-UHFFFAOYSA-N 0.000 claims description 2
- VZUFSMBGWBLOCB-UHFFFAOYSA-N C3-oxacyanine cation Chemical compound O1C2=CC=CC=C2[N+](CC)=C1C=CC=C1N(CC)C2=CC=CC=C2O1 VZUFSMBGWBLOCB-UHFFFAOYSA-N 0.000 claims description 2
- CLDZYSUDOQXJOU-UHFFFAOYSA-M C5-oxacyanine Chemical compound [I-].O1C2=CC=CC=C2[N+](CC)=C1C=CC=CC=C1N(CC)C2=CC=CC=C2O1 CLDZYSUDOQXJOU-UHFFFAOYSA-M 0.000 claims description 2
- BQRGNLJZBFXNCZ-UHFFFAOYSA-N calcein am Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)=C(OC(C)=O)C=C1OC1=C2C=C(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(=O)C)C(OC(C)=O)=C1 BQRGNLJZBFXNCZ-UHFFFAOYSA-N 0.000 claims description 2
- XVLXYDXJEKLXHN-UHFFFAOYSA-M dioc6 Chemical compound [I-].O1C2=CC=CC=C2[N+](CCCCCC)=C1C=CC=C1N(CCCCCC)C2=CC=CC=C2O1 XVLXYDXJEKLXHN-UHFFFAOYSA-M 0.000 claims description 2
- TUFFYSFVSYUHPA-UHFFFAOYSA-M rhodamine 123 Chemical compound [Cl-].COC(=O)C1=CC=CC=C1C1=C(C=CC(N)=C2)C2=[O+]C2=C1C=CC(N)=C2 TUFFYSFVSYUHPA-UHFFFAOYSA-M 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims 3
- 230000002900 effect on cell Effects 0.000 claims 2
- 239000007788 liquid Substances 0.000 abstract description 4
- 230000010261 cell growth Effects 0.000 abstract description 2
- 230000008263 repair mechanism Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 89
- VYZAMTAEIAYCRO-BJUDXGSMSA-N Chromium-51 Chemical compound [51Cr] VYZAMTAEIAYCRO-BJUDXGSMSA-N 0.000 description 28
- 238000011282 treatment Methods 0.000 description 17
- 238000001514 detection method Methods 0.000 description 11
- 150000001875 compounds Chemical class 0.000 description 9
- 238000011534 incubation Methods 0.000 description 9
- 239000003795 chemical substances by application Substances 0.000 description 7
- 206010028980 Neoplasm Diseases 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 6
- 108020004414 DNA Proteins 0.000 description 5
- 238000003026 viability measurement method Methods 0.000 description 5
- -1 Dakarbazin Chemical compound 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 4
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 4
- 230000001640 apoptogenic effect Effects 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 230000001472 cytotoxic effect Effects 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 230000002147 killing effect Effects 0.000 description 3
- 210000000822 natural killer cell Anatomy 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 231100000167 toxic agent Toxicity 0.000 description 3
- 239000003440 toxic substance Substances 0.000 description 3
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- MDKCFLQDBWCQCV-UHFFFAOYSA-N benzyl isothiocyanate Chemical compound S=C=NCC1=CC=CC=C1 MDKCFLQDBWCQCV-UHFFFAOYSA-N 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000002784 cytotoxicity assay Methods 0.000 description 2
- 231100000263 cytotoxicity test Toxicity 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 239000012636 effector Substances 0.000 description 2
- 230000005672 electromagnetic field Effects 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 230000005865 ionizing radiation Effects 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 230000031942 natural killer cell mediated cytotoxicity Effects 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 230000004962 physiological condition Effects 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- 108700012359 toxins Proteins 0.000 description 2
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-VTZDEGQISA-N 0.000 description 1
- VNFYMAPAENTMMO-UHFFFAOYSA-N 5-chloro-2-methylquinoline Chemical compound ClC1=CC=CC2=NC(C)=CC=C21 VNFYMAPAENTMMO-UHFFFAOYSA-N 0.000 description 1
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 1
- 208000030507 AIDS Diseases 0.000 description 1
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 1
- 239000004255 Butylated hydroxyanisole Substances 0.000 description 1
- GAGWJHPBXLXJQN-UORFTKCHSA-N Capecitabine Chemical compound C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1[C@H]1[C@H](O)[C@H](O)[C@@H](C)O1 GAGWJHPBXLXJQN-UORFTKCHSA-N 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 108020004638 Circular DNA Proteins 0.000 description 1
- 206010010144 Completed suicide Diseases 0.000 description 1
- 108010062580 Concanavalin A Proteins 0.000 description 1
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 1
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 1
- HTIJFSOGRVMCQR-UHFFFAOYSA-N Epirubicin Natural products COc1cccc2C(=O)c3c(O)c4CC(O)(CC(OC5CC(N)C(=O)C(C)O5)c4c(O)c3C(=O)c12)C(=O)CO HTIJFSOGRVMCQR-UHFFFAOYSA-N 0.000 description 1
- QTANTQQOYSUMLC-UHFFFAOYSA-O Ethidium cation Chemical class C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 QTANTQQOYSUMLC-UHFFFAOYSA-O 0.000 description 1
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 206010020843 Hyperthermia Diseases 0.000 description 1
- XDXDZDZNSLXDNA-TZNDIEGXSA-N Idarubicin Chemical compound C1[C@H](N)[C@H](O)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2C[C@@](O)(C(C)=O)C1 XDXDZDZNSLXDNA-TZNDIEGXSA-N 0.000 description 1
- XDXDZDZNSLXDNA-UHFFFAOYSA-N Idarubicin Natural products C1C(N)C(O)C(C)OC1OC1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2CC(O)(C(C)=O)C1 XDXDZDZNSLXDNA-UHFFFAOYSA-N 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102000008070 Interferon-gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- GQYIWUVLTXOXAJ-UHFFFAOYSA-N Lomustine Chemical compound ClCCN(N=O)C(=O)NC1CCCCC1 GQYIWUVLTXOXAJ-UHFFFAOYSA-N 0.000 description 1
- 229910052765 Lutetium Inorganic materials 0.000 description 1
- 206010027439 Metal poisoning Diseases 0.000 description 1
- 108020005196 Mitochondrial DNA Proteins 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 1
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- QAADZYUXQLUXFX-UHFFFAOYSA-N N-phenylmethylthioformamide Natural products S=CNCC1=CC=CC=C1 QAADZYUXQLUXFX-UHFFFAOYSA-N 0.000 description 1
- 102000015336 Nerve Growth Factor Human genes 0.000 description 1
- 108010025020 Nerve Growth Factor Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 101710098940 Pro-epidermal growth factor Proteins 0.000 description 1
- FOCVUCIESVLUNU-UHFFFAOYSA-N Thiotepa Chemical compound C1CN1P(N1CC1)(=S)N1CC1 FOCVUCIESVLUNU-UHFFFAOYSA-N 0.000 description 1
- 102000006747 Transforming Growth Factor alpha Human genes 0.000 description 1
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 1
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 1
- 101800004564 Transforming growth factor alpha Proteins 0.000 description 1
- 241000269370 Xenopus <genus> Species 0.000 description 1
- 229910052767 actinium Inorganic materials 0.000 description 1
- 229940008075 allyl sulfide Drugs 0.000 description 1
- 229910052789 astatine Inorganic materials 0.000 description 1
- 150000003924 bisindolylmaleimides Chemical class 0.000 description 1
- 229910052797 bismuth Inorganic materials 0.000 description 1
- 229960002092 busulfan Drugs 0.000 description 1
- CZBZUDVBLSSABA-UHFFFAOYSA-N butylated hydroxyanisole Chemical compound COC1=CC=C(O)C(C(C)(C)C)=C1.COC1=CC=C(O)C=C1C(C)(C)C CZBZUDVBLSSABA-UHFFFAOYSA-N 0.000 description 1
- 229940043253 butylated hydroxyanisole Drugs 0.000 description 1
- 235000019282 butylated hydroxyanisole Nutrition 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000025084 cell cycle arrest Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229960005395 cetuximab Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 229960000975 daunorubicin Drugs 0.000 description 1
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- LDCRTTXIJACKKU-ONEGZZNKSA-N dimethyl fumarate Chemical compound COC(=O)\C=C\C(=O)OC LDCRTTXIJACKKU-ONEGZZNKSA-N 0.000 description 1
- 229960004419 dimethyl fumarate Drugs 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229960003668 docetaxel Drugs 0.000 description 1
- 229960004679 doxorubicin Drugs 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000005670 electromagnetic radiation Effects 0.000 description 1
- 210000002308 embryonic cell Anatomy 0.000 description 1
- 210000001671 embryonic stem cell Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229960001904 epirubicin Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 230000036031 hyperthermia Effects 0.000 description 1
- 230000002631 hypothermal effect Effects 0.000 description 1
- 229960000908 idarubicin Drugs 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 229960003130 interferon gamma Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 229960002247 lomustine Drugs 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- SGDBTWWWUNNDEQ-LBPRGKRZSA-N melphalan Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N(CCCl)CCCl)C=C1 SGDBTWWWUNNDEQ-LBPRGKRZSA-N 0.000 description 1
- 229960001924 melphalan Drugs 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003990 molecular pathway Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 229960001756 oxaliplatin Drugs 0.000 description 1
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 1
- QOFFJEBXNKRSPX-ZDUSSCGKSA-N pemetrexed Chemical compound C1=N[C]2NC(N)=NC(=O)C2=C1CCC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 QOFFJEBXNKRSPX-ZDUSSCGKSA-N 0.000 description 1
- 229960005079 pemetrexed Drugs 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 229960004641 rituximab Drugs 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000037351 starvation Effects 0.000 description 1
- 239000002438 stress hormone Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229960000575 trastuzumab Drugs 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 231100000747 viability assay Toxicity 0.000 description 1
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 1
- 229960004528 vincristine Drugs 0.000 description 1
- GBABOYUKABKIAF-GHYRFKGUSA-N vinorelbine Chemical compound C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC([C@]23[C@H]([C@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC GBABOYUKABKIAF-GHYRFKGUSA-N 0.000 description 1
Classifications
-
- 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
- G01N33/5014—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 for testing toxicity
Definitions
- the present invention relates to the field of biological research. More in particular, it relates to biological research utilizing living cells. Even more in particular, it relates to the measurement of the status of cells, such as the viability of cells.
- Biological research of today is to a significant fraction relying on cells as model organisms.
- the measurement of how cell status is dependant of a certain treatment is of great interest.
- Cell status may include qualities as cell growth, changes in cell morphology, triggering of repair mechanisms after damage, viability, and the like.
- One common example of cell status measurement is the viability measurement, i.e. the measurement of the ability of a cell to survive a certain treatment is of great interest.
- the reverse is also common, i.e. the measurement of how efficient a certain treatment is in killing cells.
- Yet another viability measurement of interest is to follow natural cell death (apoptosis) either through triggering of apoptotic molecular pathways (i.e. not poisoning the cell, but rather trick it to commit suicide) or as it occurs without use of molecular effectors.
- PI propidium iodide
- toxicity measurements using the antonym toxicity There are other methods available for the measurement of viability (such methods are sometimes referred to as toxicity measurements using the antonym toxicity).
- One common method relies on radioactive chromium (51Cr), which is absorbed by living cells and released when the cells die. Such measurements typically comprise an incubation period, during which the cells are loaded with 51Cr, a toxin treatment period (and optionally a waiting period) prior to harvesting the cell culture medium. Since dead cells have release their chromium into the cell culture medium, the radioactivity of the medium is proportional to the number of killed cells. Measurements relying on 51Cr are terminal and can therefore only provide information on the cell viability status at the time of harvesting.
- the object of the present invention is to provide an improved method for measuring cell status in general.
- the invention is particularly useful for time-resolved measurements of cell viability.
- the method according to the invention is defined in claim 1.
- the method for measuring cell viability uses of
- the invention in a preferred embodiment comprises three characteristic components.
- Suitable markers include (but are not limited to) radioactive markers, fluorescent markers and chemoluminiscent markers.
- Suitable cells include (but are not limited to) prokaryotic or eukaryotic cells that can be cultured in a laboratory.
- suitable cells are human cancer cell- lines, for example the cell-line A431 (ATCC, CLR 1555, Rocksville MD USA). The time resolved measurement is typically made in a device described in WO2005080967, which is incorporated by reference herein.
- Another application is the study of apoptosis and potential apoptotic inducers and apoptotic inhibitors.
- Yet another application is the development of cancer treatments, i.e. finding suitable means for killing tumor cells.
- Still another application is the study of cell viability when put under influence of external treatments, such as cell resistance to electromagnetic radiation.
- Yet another application is to study the function or effect of the immune system when killing intruding cells.
- Still another application is the measurement of total DNA in the target cells as an indirect measure of growth.
- Figure 1 shows a flow chart of the method
- Figure 2 shows a suitable instrument, known in prior art, for performing the measurement in the viability method
- Figure 3 shows results from one 51Cr incubation experiment
- Figure 4 shows results from two 51Cr incubations on untreated cells and from one 51Cr incubation on treated cells
- Figure 5 shows results from two 51Cr incubations on untreated cells, wherein the 51Cr concentration was different; and Figure 6 shows results from a series of combined 51Cr incubations and treatments.
- the term "cell status” refers to the general condition of a cell population with respect to one of more aspects including (but not limited to) DNA content, intracellular metal ions concentrations, intensity of metabolism, fraction necrotic cells, and the like.
- viability refers to the viability status of the cells subject to measurement. Viable cells are alive and growing, and poor viability is recognized as slow or absent growth, poor cell morphology, or necrosis.
- the cells attached to the solid support are denoted "target cells”.
- Target cells include (but are not limited to) pro kary otic cells, eukaryotic cells, tissue slices (thinner than 2 mm), and small organisms (less than 2mm in diameter) that can be cultured in a laboratory.
- target cells include (but are not limited to) human cancer cell lines, human embryonic stem cell lines (na ⁇ ve or differentiated), cancer cell lines from other mammals, embryonic cell lines from other mammals, insect cells and xenopus cells.
- the method requires a compound to be used for monitoring the status of the target cells, said compound being denoted "status marker”.
- status markers include, but are not limited to, radioactive markers (e.g. 51Cr, 18-F-deoxyglucose), fluorescent markers (e.g.
- the present invention aims at providing a method of measuring cell status by enabling a time-resolved detection of status markers in target cells. Due to the time-resolved detection, the effect of different chemical or environmental conditions can be detected as it appears.
- target cells are loaded with viability markers (110), followed by an optional wash (120).
- the cells are treated with the agent or exposed to a condition for which the status should be determined (130), followed by an optional wash (140).
- the last step in the method is the time-resolved detection of status marker associated with target cells (150), which is a direct or indirect measure of the status of the cells.
- cytotoxic drugs e.g. Doxorubicin, Daunorubicin, Fluorouracil, Tiotepa, Hydroxykarbamid, Karboplatin, Metotrexat, Etoposid, Paklitaxel, Vinkristin, Dakarbazin, Epirubicin, Oxaliplatin, Lomustin, Cytarabin, Vinorelbin, Docetaxel, Pemetrexed, Topotekan, Amsakrin, Merkaptopurin, Busulfan, Melfalan, Metotrexat, Idarubicin, Mitoxantron, Mitomycin, or Capecitabin) .
- cytotoxic drugs e.g. Doxorubicin, Daunorubicin, Fluorouracil, Tiotepa, Hydroxykarbamid, Karboplatin, Metotrexat, Etoposid, Paklitaxel, Vinkristin, Dakarbazin, Epirubicin, Oxaliplatin,
- cytotoxic proteins e.g. monoclonal antibodies like Rituximab, Trastuzumab, and Cetuximab
- growth factors e.g. EGF, NGF, TGF-alpha, TGF-beta
- other proteins e.g. insulin
- detergents e.g. tween 40, tween 20, Triton XlOO, octylglycoside or NP40
- inorganic compounds or metals e.g. Copper, Lead, Cadmium, or Mercury
- radioactive compounds e.g.
- Possible conditions include (but are not limited to) starvation, hyperthermia, hypothermia, increased or decreased ionic strength compared to physiological conditions, increased or decreased pH compared to physiological conditions, non-ionizing radiation, ionizing radiation, static electromagnetic fields, fluctuating electromagnetic fields (e.g. microwave treatment), and mechanical treatment (e.g.
- apoptosis- inducing effector compounds can be used, including (but not limited to) bisindolylmaleimide compounds (as defined in US6284783, which is incorporated by reference herein), butylated hydroxyanisole, allyl sulfide, benzyl isothiocyanate, and dimethyl fumarate.
- the method can be conducted in slightly different ways.
- One other example is as described in figure Ib, wherein the treatment of the target cells could be the first step (111), followed by an optional wash (121), after which the treated target cells are loaded with status marker (131). After an optional wash (141), status marker is detected using a time-resolved detection technology.
- FIG. 1c Yet another example is as described in figure Ic, wherein the treatment of the target cells and the loading of status marker is performed simultaneously (112), followed by an optional wash (122). Next, status marker is detected using a time-resolved detection technology (132).
- Still another example is as described in figure Id, wherein the first step is the treatment of the target cells (113), followed by an optional wash (123). Next, status marker is loaded and simultaneously detected using a time- resolved detection technology (133).
- FIG. 1e Yet another example is as described in figure Ie, wherein the first step is the loading of status marker during time- resolved detection (114), followed by an optional wash (124). Next, the treatment of the cells is started and the effect of it being simultaneously detected using a time-resolved detection technology (134).
- the preferred method for completing steps 114, 150, 151, 132, 133, and 134 in figure 1, i.e. the detection, has been previously disclosed [WO2005080967, which is incorporated by reference herein] and is schematically described in figure 2.
- the method relies on target cells (202) being attached to a defined area on a solid support (201).
- a reference area in this case opposite to the target area.
- a liquid is in contact with the solid support to enable a suitable environment for the target cells.
- the liquid may possibly contain status marker, so that the loading of target cells takes place during detection.
- the solid support is inclined and slowly rotated using a motor (203).
- a detector capable of detecting the label attached to the species used is mounted (204) over the elevated portion of the solid support.
- an elevated signal will be registered in case the status marker has bound to or is present in the target cells.
- the rate of change of status marker density can be followed by depicting the difference between the detected signal from target area and reference area over time.
- target cells are attached to a selected portion of a solid support
- the present invention makes it possible to follow cell status in real time, thereby monitoring the progress of processes toxic to the cell.
- step 113 and 123 were ignored and the baseline measurement started at step 133.
- a time-resolved measurement was performed using the preferred device as described in figure 2, using a detector sensitive to gamma radiation in the energy range 100- 1000 keV.
- the resulting graph of amount of 51Cr versus time is shown in figure 3. As seen, the cells absorb 51Cr during approximately 16 hours after which the 51Cr is declining.
- the baseline measurement was repeated in order to verify the stability of the method.
- target cells were treated with Agent during one hour (113), followed by a wash (123) and approximately 15 hours after Agent treatment, the target cells were loaded with 51Cr during measurement (133).
- the resulting curve of amount of 51Cr versus time is shown in figure 4, together with the two baseline curves.
- Figure 4 shows that the decline of 51Cr is delayed in the treated cells compared to the baseline measurement. Furthermore, the results from the two baseline measurements are in agreement, indicating that the viability measurement is robust.
- a probable cause for the general shape of the curves in figure 3 and figure 4 is that the target cells might be sensitive to 51Cr itself.
- the decline of 51Cr appeared earlier at higher concentration of 51Cr as shown in figure 4.
- the fact that the cytotoxic Agent is shifting the decline towards later points in time indicates that Agent is forcing target cells into cell-cycle arrest, because dividing cells are likely more sensitive to metal poisoning than static cells.
- Example 2 The method described above (figure 1, steps 114-124- 134) was tested with target cells of type A431 (a human cancer cell-line) grown on approximately one quarter of a 10cm circular cell-dish.
- the cellular status detected in this example was viability and the status marker was 51 -chromium.
- the cells While detecting the 51Cr level in the cells, the cells were incubated with 51Cr during approximately 2h. After incubation, a toxic agent (Triton XlOO) was added to the cell dish. The release of 51Cr was then followed approximately 7 hours. The procedure was repeated several times with varying concentrations of the toxic agent Triton XlOO.
- Triton XlOO Triton XlOO
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Hematology (AREA)
- Toxicology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
L'invention concerne un procédé pour la mesure de l'état cellulaire, à savoir des qualités telles que la croissance cellulaire, des changements de la morphologie de la cellule, le déclenchement de mécanismes de réparation après endommagement, la viabilité et similaires. Le procédé comprend la fixation de cellules cibles sur un support solide, le marquage de la cellule cible par un marqueur d'état (par exemple, une molécule fluorescente ou chimioluminescente ou un émetteur de rayonnement radioactif), et la détection de la quantité de marqueur d'état en résolution temporelle. L'invention concerne en outre un tel procédé de mesure, consistant à mettre le support solide étant en contact avec un liquide, puis à effectuer une mesure permettant détecter la présence d'un marqueur d'état, ainsi qu'une mesure de référence. Le liquide est temporairement retiré pendant la mesure.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
SE0701925 | 2007-08-28 | ||
SE0701925-0 | 2007-08-28 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2009029039A1 true WO2009029039A1 (fr) | 2009-03-05 |
Family
ID=40387562
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/SE2008/050974 WO2009029039A1 (fr) | 2007-08-28 | 2008-08-28 | Procédé de mesure de la viabilité d'une cellule |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2009029039A1 (fr) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8993259B2 (en) | 2012-05-02 | 2015-03-31 | Charles River Laboratories, Inc. | Method of viability staining with membrane permeable fluorescent dye and membrane impermeable fluorescence quencher |
EP3077814A4 (fr) * | 2013-11-04 | 2017-06-07 | Charles River Laboratories, Inc. | Procédé de détection de cellules viables dans un échantillon de cellules |
US9709500B2 (en) | 2012-05-02 | 2017-07-18 | Charles River Laboratories, Inc. | Optical method for detecting viable microorganisms in a cell sample |
US10324036B2 (en) | 2012-05-02 | 2019-06-18 | Charles River Laboratories, Inc. | Porous planar cell capture system |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005080967A1 (fr) * | 2004-02-20 | 2005-09-01 | Ridgeview Instruments Ab | Procede et dispositif pour la caracterisation des interactions entre differentes especes |
-
2008
- 2008-08-28 WO PCT/SE2008/050974 patent/WO2009029039A1/fr active Application Filing
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005080967A1 (fr) * | 2004-02-20 | 2005-09-01 | Ridgeview Instruments Ab | Procede et dispositif pour la caracterisation des interactions entre differentes especes |
Non-Patent Citations (3)
Title |
---|
BRUNNER K.T. ET AL: "Quantitative assay of the lytic action of immune lymphoid cells on 51-Cr-labelled allogeneic target cells in vitro; inhibiton by isoantibody and by drugs", IMMUNOLOGY, vol. 14, no. 2, 1968, pages 181 - 196, XP003025141 * |
MUELLER H. ET AL: "Comparison of the usefulness of the MTT,ATP and calcein assays to predict the potency of cytotoxic agents in various human cancer cell lines", JOURNAL OF BIOMOLECULAR SCREENING, vol. 9, no. 6, 2004, pages 506 - 515, XP009081506 * |
PROVINCIALI M. ET AL: "Optimization of cytotoxic assay by target cell retention of the fluoroscent dye carboxyfluoroscein diacetate (CFDA) and comparison with conventional chromium-51 release assay", JOURNAL OF IMMUNOLOGICAL METHODS, vol. 155, 1992, pages 19 - 24, XP023657846 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8993259B2 (en) | 2012-05-02 | 2015-03-31 | Charles River Laboratories, Inc. | Method of viability staining with membrane permeable fluorescent dye and membrane impermeable fluorescence quencher |
US8993260B2 (en) | 2012-05-02 | 2015-03-31 | Charles River Laboratories, Inc. | Fluorescence-based viability staining method using a membrane permeable flourescent dye and membrane impermeable fluorescence quencher |
US9709500B2 (en) | 2012-05-02 | 2017-07-18 | Charles River Laboratories, Inc. | Optical method for detecting viable microorganisms in a cell sample |
US10324036B2 (en) | 2012-05-02 | 2019-06-18 | Charles River Laboratories, Inc. | Porous planar cell capture system |
US10976258B2 (en) | 2012-05-02 | 2021-04-13 | Charles River Laboratories, Inc. | Porous planar cell capture system and method of use |
EP3077814A4 (fr) * | 2013-11-04 | 2017-06-07 | Charles River Laboratories, Inc. | Procédé de détection de cellules viables dans un échantillon de cellules |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Nuccitelli et al. | Nano-Pulse Stimulation is a physical modality that can trigger immunogenic tumor cell death | |
Bhargava et al. | Fate of nanoplastics in marine larvae: a case study using barnacles, Amphibalanus amphitrite | |
Stephens et al. | Development of apoptosis in irradiated murine tumors as a function of time and dose | |
Sukkurwala et al. | Screening of novel immunogenic cell death inducers within the NCI mechanistic diversity set | |
Mendez-Vilas et al. | Microscopy: Science, technology, applications and education | |
WO2009029039A1 (fr) | Procédé de mesure de la viabilité d'une cellule | |
Kozlova et al. | Opposite effects of electroporation of red blood cell membranes under the influence of zinc ions. | |
Joksić et al. | Size of silver nanoparticles determines proliferation ability of human circulating lymphocytes in vitro | |
CN110662843A (zh) | 细胞的膜电位/膜电流的测定方法 | |
EP3430398A1 (fr) | Localisation sous-cellulaire d'analytes cibles | |
CN111886331A (zh) | 一种保存粪便的方法 | |
Christiansen et al. | The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage | |
Nastulyavichus et al. | Dynamic light scattering detection of silver nanoparticles, food pathogen bacteria and their bactericidal interactions | |
Skamrova et al. | Influence of mobile phone radiation on membrane permeability and chromatin state of human buccal epithelium cells | |
Pejchal et al. | Cytokinesis-block micronucleus (CBMN) assay/CBMN cytome assay in human lymphocytes after in vitro irradiation and its use in biodosimetry | |
Stanevičienė et al. | Subacute effects of cadmium and zinc ions on protein synthesis and cell death in mouse liver | |
Singh et al. | Alpha-tocopherol succinate-mobilized progenitors improve intestinal integrity after whole body irradiation | |
Punshon et al. | High-resolution elemental mapping of human placental chorionic villi using synchrotron X-ray fluorescence spectroscopy | |
CN115266664B (zh) | 一种免疫细胞治疗制剂体外杀伤效力评价方法及其应用 | |
Tirosh et al. | Immune cytolysis viewed as a stimulatory process of the target | |
Dong et al. | The role of SIRT1 in the process of Toxoplasma gondii infection of RAW 264.7 macrophages | |
Gholampour et al. | Chronic exposure to extremely low frequency electromagnetic field induces mild renal damages in rats | |
Shckorbatov et al. | Electromagnetic field effects on Artemia hatching and chromatin state | |
Ren et al. | Single cell elemental analysis using nuclear microscopy | |
Heidari Zare et al. | Microbiological investigation of an antibacterial sandwich layer system |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 08828512 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 08828512 Country of ref document: EP Kind code of ref document: A1 |