Cov ntsiab lus: OECD 471, OECD 473, OECD 476, OYECD 487, MENSION TEST SHERT, MEs TEST SHERT, HPRT / HGPRT ASSAY, TK ASSAYY, TK ASSAY
Iphas khoom
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Khoom |
Kev qhia tshwj xeeb |
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Ntxias lub siab s9 cov khoom lag luam |
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Iphase nas (Sprague - Dawb) Mob siab s9, induction, txiv neej |
35mg / ml, 1ml |
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Iphase nas (Sprague - Dawb) Mob siab s9, induction, txiv neej |
35mg / ml, 2ml |
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Iphase nas (Sprague - Dawb) Mob siab s9, induction, txiv neej |
35mg / ml, 5ml |
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35mg / ml, 1ml |
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35mg / ml, 5ml |
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Genotoxicity soj ntsuam cov khoom siv |
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100/150/200/250 TAUNCH |
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6well * 24 / 6well * 40 |
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| Iphase hauv cov vitro mammalian cell micronucleus kuaj | 5ml * 32 xeem |
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16 * 96 WELS / 4 * 384 WELLS |
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96 zoo |
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20ml * 36 xeem |
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20ml * 36 xeem |
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Iphas nyob rau hauv - vitro chromosome aberration ntsuas cov khoom siv |
5ml * 30 xeem |
Lub hauv paus rau phau ntawv
Kev tsis pub tus nas lub siab s9yog qhov tseem ceeb ntawm kev ntsuas covKev tiv thaiv caj cesmuaj peev xwm ntawm cov tshuaj lom neeg, tshwj xeeb tshaj yog nyob rau hauv kev tswj hwm toxicology. Nws yog feem ntau siv nyob rau hauv kev sib xyaw ua ke nrog nyob rau hauv vitro saib, xws li lubAmes xeem thiabkev sim sib hloov, txhawm rau ntsuas cov khoom mutagenic ntawm cov tebchaw. Los ntawm kev muab Enzymatic system Rich nyob cytochrome P450 (CYP450), cov txheej txheem sib npaug hauv kev sim siab tias cov tshuaj lom neeg tuaj yeem ua rau muaj kev sib cav sib ceg lossis mob qog noj ntshav.
Kev tsis pub tus nas lub siab s9
Lub siab s9 feem kev hais txog ib qho post - mitochondrial supernatant tau los ntawm cov rativen hom tom qab muaj kev sib tw ntawm centrifugation Lo lus "ntxias" yog hais txog kev kho ntawm cov nas uas muaj cov kev ua kom muaj cov kev ua kom zoo nkauj, tshwj xeeb yog lub luag haujlwm rau cov metabolism hauv cov xenobiotic.
Qhov kev ua kom muaj lub siab s9 feem s9 feem muaj ntau cov enzymes uas tau koom nrog theem I thiab theem II metabolism ntawm cov tshuaj lom neeg. Qhov no suav nrog cov enzymes xws li Cytochrome P450 uas pab txhawb kev ua haujlwm ntawm kev ua yeeb yam, yog li ua kom tib neeg lub siab metabolism.
CYR450 Kev Ua Si thiab Kev Ua Haujlwm Metabolic
Cytochrome P450 enzyme tsev neeg (CYP450) ua ntau cov tshuaj, suav nrog ntau cov tshuaj, xws li cov tshuaj ib puag ncig, thiab carcinogens. Qhov kev ua kom muaj lub siab s9 feem muaj cov enzymes thiab yog qhov tseem ceeb rau kev ntsuas cov tshuaj uas tsuas yog ua genotoxic tom qab kev ua haujlwm metosic tom qab ntsuas me me.
Ntau cov tebchaw tau pib tsis yog - toxic tab sis tuaj yeem dhau los ua kom lom tom qab metabolism hauv lub siab. Cov pro - mutagens (uas yuav tsum tau meabolic ua kom ua mutagenic) thiab pro - carcinogens (uas yuav tsum tau kuaj xyuas los ntawm cov khoom siv s9 metabolic ua kom haum. Yog tsis muaj qhov induction ntawm cov metabolic enzymes zoo li cy450, cov tshuaj no yuav tsis muaj kev puas tsuaj hauv cov qauv genotoxicity kev ntsuas.
Los ntawm kev ntxiv cov fiver nas s9 rau hauv cov chaw tshawb fawb, cov kws tshawb nrhiav tuaj yeem txheeb xyuas yuav ua li cas cov tshuaj uas cuam tshuam nrog CYP450 enzymes. Qhov no yog qhov tshwj xeeb tshaj rau cov tshuaj, raws li cov metabolism los ntawm cy450 enzymes yog ib qib tseem ceeb hauv cov tshuaj.
Cov ntawv thov ntawm cov nas daim siab s9
- 1. Ames xeem (OECD 471)
Cov Ames xeem, teev rau hauv cov lus qhiaOECD 471, yog ib qho ntawm cov zoo tshaj plaws - Paub thiab cov hau kev siv rau kev ntsuas kev sib cav sib tua. Nws cuam tshuam nrog cov kab mob Salmonella rau cov tshuaj ntsuam xyuas seb nws puas tsim kev hloov pauv tau tias nws ua rau cov kab mob rov qab mus rau histidine - Independent State.
Txhawm rau sim saib tib neeg cov txheej txheem metabolic, ua rau cov nas lub siab s9 yog feem ntau ntxiv rau qhov ntsuas ntsuas. Lub S9 feem muab cov enzymes tsim nyog, suav nrog CYP450, uas tej zaum yuav tau rau metabolize lub compound ua kev sib hloov hauv cov kab mob DNAs. Qhov kev sib txuam ntawm S9 nrog cov Ames xeem tso cai rau kev ntsuas kev hloov pauv ntawm kev ua kom pom kev ncaj qha los ntawm kev ua kom pom kev sib tsoo thiab kev ua kom zoo nkauj mutagenic.
- 2.Kev sim sib hloov pauv (Chromosomal ntsuas, micronucleus kuaj, thiab lwm yam hauv vitro presays)
Ntxiv rau kev kuaj Ames, kev sib hloov yuav siv tau siv los ntsuas qhov muaj peev xwm ntawm cov tshuaj lom neeg. Cov kev ntsuam xyuas no tau muaj ntau dua hauv kev txheeb xyuas qhov ntau ntawm caj ces, suav nrog cov kev sib hloov caj ces, suav nrog cov kev sib hloov pauv ntawm caj ces, thiab cov kev sib hloov pauv ntawm micronuclei. Kev tsis pub muaj lub siab s9 yog siv hauv kev sib hloov pauv rau cov laj thawj hauv qab no:
Micronucleus xeem (OECD 487)
Cov kev sim no kuaj pom kev tsim ntawm micronuclei hauv cov hlwb, uas yog cov chromosomes lossis tag nrho cov chromosomes uas tsis tau ua incorporated rau hauv lub nucleus thaum lub sijhawm faib khoom. Tus Micronucleus kuajtuaj yeem tshawb xyuas clastogenic (chromosome - Breaking) thiab aneugenic (cuam tshuam chromosome tooj) teebmeem. Kev tsis txaus siab river s9 yog siv rau ntawm no kom qhib cov quav yeeb tshuaj metabolically, raws li qee cov tshuaj chromosomal puas tsuaj thaum lawv tau mob siab los ntawm daim siab enzymes. S9 feem kev txhim kho cov kev nkag siab ntawm kev sim los ntawm kev simulating cov kev ua kom metabolic uas yuav tshwm sim hauv vivo.
Chromosomal Kev Ntsuas (OECD 473)
Qhov kev sim ntsuas no seb puas yuav ua rau cov teeb meem kev puas tsuaj rau cov chromosomes los ntawm kev ua txhaum cai, tshem tawm, kev txhais lus, lossis lwm hom kev ua haujlwm aberrations. Zoo li micronucleus kuaj, tuschromosomal xeemtuaj yeem siv nrog lossis tsis muaj kev ua kom muaj teeb meem. Kev tsis pub tus nas lub siab s9 yog ntxiv rau qhov ntsuas ntsuas kom pom cov txheej txheem metabolic hauv lawv daim ntawv qub tab sis yuav ua tau li tom qab daim siab metabolism.
Rooj 1. Kev txiav txim siab sib piv ntawm kev sib tw
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Yam zoo |
HPRT / HGPRT ASSAY |
L5178Y TK assay |
Hu TK assay |
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Gene hom phiaj |
~ 650 BP Coding cheeb tsam |
~ 1,200 bp exon / intron cheeb tsam |
~ 1,000 BP Coding cheeb tsam |
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Tom qab mf |
~ 1-5 × 10⁻⁶ |
~ 1-5 × 10⁻⁵ |
1-3 × 10⁻⁶ |
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Cov Hom Ntawv Kawg |
Taw tes sib hloov nkaus xwb |
Taw tes + Chromosomal aberrations |
Taw tes sib hloov nkaus xwb |
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Colony Morphology |
Cev khaub ncaws tseem ceeb |
ME VS. CONGIES CONGIES |
Cev khaub ncaws tseem ceeb |
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Kev tswj cov lus qhia |
OECD 476 |
OECD 490 |
OECD 476 |
HPRT / HGPRT ASSAY (OECD 476)
Nyob rau hauvHPRT / HGPRT ASSAY, Kab lis kev cai ntawm Suav hamster (cho lossis tib neeg lymphobloid (uas tau muab cov tshuaj sim hauv cov nas uas yuav tsum tau hloov kho pro-mutagens mus rau hauv DNA-reactive hom. Tom qab ib qho kev kho mob me me thiab muaj hnub nyoog xya xyoo, cov hlwb muaj qhov sib tw nrog 6-thioguanine; Tsuas yog clones cov kabmob poob-ntawm-ua haujlwm hloov pauv hauv hypoxanthine-guanine phosphoribosyltretase noob muaj sia nyob. Los ntawm kev sib piv mutant colony suav rau tag nrho cov volony, cov kws tshawb nrhiav txiav txim siab ntau zaus txog kev tswj hwm thiab keeb kwm kev tswj hwm, qhia txog Genotoxic muaj peev xwm.
TK prests (OECD 490 & OECD 476)
TusTk assay ntiav ua L5178y nas Lympoma Hlwb (OECD 490) lossis CIA CO CANS (OECD 476) Tus tsim tawm heterozygous ntawm thymidine Kinase Locus. Tom qab raug tshuaj lom neeg ± S9 sib tov thiab yog plated nyob rau hauv nruab nrab uas muaj lub trifluorothernymidine, uas tua TK-profess hlwb. Ciaj sia Tk⁻ mutants tsim cov kob xim ntau tshaj 10-14 hnub, nrog me-colony dissants feem ntau xav txog cov xwm txheej chromosomal feem ntau muaj kev sib raug zoo. Raws li nyob rau hauv HPRT / HGPR ASSAY, nruj nkaus li Cytotoxicity tswj thiab zoo mutagen shours
- 4. Kev Ntsuam Xyuas Kev Ntsuas Mob Cancer
Kev tsis pub muaj lub siab s9 tseem siv rau hauv kev tshawb fawb kom ntsuas cov mob cancer - ua rau muaj peev xwm ntau yam. Los ntawm simulating cov txheej txheem metabolic uas tshwm sim hauv tib neeg lub siab, tus S9 feem kev sib tw uas yuav ua rau muaj kev sib txuas ntawm cov khoom siv uas yuav ua rau mob qog noj ntshav.
Txhim Kho Ames Kev Ntsuas Nrog Indive Hamster Liiver S9
Raws li cov lus qhia tshuaj European tau muab los ntawm European tshuaj chaw haujlwm (EMA), cov kev ntsuas ames tsis txaus ntseeg, tshwj xeeb yog nitrosylamine (ndma), thiab lwm tus. Yog li ntawd, txoj kev ntsuas ames txhim kho, tsim los ntawm National Center forxicical kev tshawb fawb (NCTR), ib qho kev faib tawm ntawm U.S. Kev Tswj Xyuas Khoom Noj (FDA), tau raug pom zoo raws li txoj kev xaiv tau zoo dua. Qhov kev sim ua tau zoo tshaj plaws Ames ua rau pom kev sib tw hamster lub siab s9, uas ua rau muaj 30% nas lub siab s9 thiab 30% hamster daim siab s9. Rat thiab Hamster Desmosomal supernatants (S9S) yuav tsum npaj los ntawm nas tsuag kho nrog Cytochrome P450 enzyme - Txhaum tshuaj. Los ntawm kev siv hamster lub siab s9, txoj kev txhim kho txhim kho zoo dua li simulates tib neeg cov metabolism thiab nce kev ntseeg tau ntawm cov txiaj ntsig.
Tag
Kev ua kom muaj lub siab s9 ua cov cuab yeej tseem ceeb hauv kev sim siab tshaj plaws hauv Genotoxicity Testing, pab cov chaw tshawb nrhiav thiab cov chaw tswj hwm ntsuas cov tshuaj tua kabmob thiab carcinogenic muaj peev xwm ntawm cov tshuaj lom neeg. Los ntawm kev muab cov kev ua kom metabolic ua kom muaj zog, nws txhim kho lub peev xwm zoo li cov kev xeem ames, thiab CYP450 kev sim ua kom tau cov khoom siv metabolic los ua genotoxic. Nws cov ntawv thov yog qhov tseem ceeb rau kev ua kom muaj kev nyab xeeb ntawm cov tshuaj tshiab, tshuaj lom neeg, thiab cov neeg siv khoom. Nrog nws lub luag haujlwm hauv kev simulating tib neeg lub siab metabolism, qhov ua rau cov nas lub siab s9 txuas ntxiv mus rau qhov tseem ceeb ntawm kev ntsuas kev nyab xeeb niaj hnub thiab kev tswj hwm kev nyab xeeb kev nyab xeeb.
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