Thawj Hepatocytes: cov cuab yeej tseem ceeb rau kev nce qib tsis yog (tshuaj ntsuam hauv cov kev tshawb fawb vitro

Thawj Hepatocytes: cov cuab yeej tseem ceeb rau kev nce qib tsis yog (tshuaj ntsuam hauv cov kev tshawb fawb vitro

Mob siab, raws li thawj qhov khoom nruab nrog rau daim tshuaj rau tshuaj, ua si lub luag haujlwm tseem ceeb hauv cov tshuaj metabolism thiab cov txheej txheem lom. Thawj Hepatocytes muaj cov spectrum tag nrho ntawm cov yam ntxwv ntawm cellular thiab cytochrome p450 (suav nrog cov kab ke cytochymes, suav nrog cov kab mob metosulum Vim li no, thawj tus neeg Hepatocytes tau pom tias yog tus qauv kub los tsim cov kws tshawb fawb hauv kev siv tshuaj yeeb tshuaj, thiab kev tshawb xyuas toxicity. Kab lus no muab cov ntsiab lus ntawm 2D thiab cov qauv 3D haiv neeg raws li cov Hepatocytes thiab lawv cov ntawv thov hauv kev tsim tshuaj.

Cov Ntsiab Lus: Tus Thawj Coj Hepatocytes, 2D cultivation, 3D sau qoob, 3D sau qoob, engodegs, co - Kab lis kev cai.

1. 1. Kev rho tawm ntawm Thawj Hepatocytes

Kev cais ntawm Gepatocytes tseem ceeb heev hauv kev tsim cov qauv siab tseem ceeb, nrog ob kauj ruam collagenase txoj kev yog feem ntau siv. Nyob rau hauv cov tsiaj me, nplooj siab ua tau ntawm Portal txoj leeg los yog qis dua tsiaj feem ntau yuav tsum muaj qhov ua kom zoo los yog ntu. Ob peb yam tseem ceeb cuam tshuam rau kev vam meej ntawm Hepatocytes: Ua ntej, collagenase yuav tsum tsis yog - cyotoxic. Thib ob, lub sijhawm ntawm kev zom zaub mov yog qhov tseem ceeb-Ob qho tib si hauv qab - kev zom thiab dhau - zom tuaj yeem sib haum hapatocyte tawm los thiab vibility. Qhov thib peb, lub siab mob yuav tsum yog qhov zoo tshaj plaws, raws li hepatocytes yog qhov rhiab heev rau ischemic kev puas tsuaj. Lub siab siv rau kev npaj Hepatocyte yuav tsum txo cov nqi me me kom txo qis metabolic tus nqi thiab tiv thaiv cov metabolic hypoxia thiab tom qab ischemia.

Cov qauv rau thawj hepatocytes siv hauv kev tshawb fawb hauv tshuaj yog raws li hauv qab no:

1. 2. Thawj Hepatocyte 2D Kab lis kev cai

Tus tshuam Hepatocyte qauv

Nws muaj cov tshuaj tiav tiav lawm - metabolizing enzymes thiab cofactors, ua rau nws kawm txog kev kawm ntau yam kev nthuav dav. Txawm li cas los xij, lub vavility ntawm ncua kev ncua ntawv hepatocytes thiab cov kev ua yeeb yam ntawm cov tshuaj - metabolizing maj mam nce, txwv kom lub zog loj dua, txwv tsis pub dhau 4 teev. Cov qauv no feem ntau yog siv los kwv yees cov tshuaj nrog kev muaj pes tsawg tus nqi siab. Thaum qhov kev tshem tawm tus nqi yog tsawg dua 20%, kev tshem tawm qhov tseeb tsis tuaj yeem txiav txim siab. Ncua Kev Kawm Ntawv Hepatocyte - Raws li nyob rau hauv Vitro Metabolism qauv yog qhov tsis txaus ntseeg rau cov khoom siv metabolic, yog li txo lawv lub peev xwm los twv seb cov khoom lag luam me me thiab cov khoom lag luam metabolic ntawm cov tebchaw no. Kev ncua sij hawm Kev Kawm Ntawv Hepatocyte Relay Txuj Ci (Daim duab 1) tuaj yeem siv los txuas ntxiv lub sijhawm ua kom ntev txog 20 teev lossis ntev dua.

Daim Ntawv Thov:Enzyme kev ua si thiab metabolic stability kev kawm rau cov tshuaj me me tshuaj.

Daim Duab 1. Ncua tseg Hepatocyte Relay Txuj Kev Hloov Cov Txheej Txheem

Tau qhov twg los: Kev Siv Tshuaj Yeeb Tshuaj, Kev Siv Tshuaj Yeeb, 2012,40 (9): 1860-1865

Lub platable hepatocytes qauv

Thawj Hepatocytes yog kev coj ua hauv 2D system ntawm collagen - coated kab lis kev cai daim hlau. Cov hepatocytes nthuav tawm cov epithelial morphology, nrog kev tawm tsam nuclei, feem ntau nthuav tawm hauv daim ntawv binucleed. Lub drawbacks ntawm ib tus hepatocyte monolayer kab lis kev cai muaj xws li: 1. Kev hloov kho ntawm tes polarity thiab Function.2. Tsis muaj lwm hom cell tsuas yog (i.e., tsis yog - parenchymal cell) uas yuav tsum muaj rau ib txwm muaj nuj nqi.3. Kev tsis muaj peev xwm muab cov as -ti as thiab paracrine txaus los txhawb nqa hepatocytes hauv kev ua lawv cov haujlwm (xws li kua roj ntsha (serum protein).

Cov Ntawv Thov:

1). Kev ntsuam xyuas cov tshuaj - Cov Kev Sib Tham Cov Tshuaj:Qhov no suav nrog enzyme induction, enzyme inhibition, thiab cov kev tshawb fawb txuas ntxiv. Firstly, thaum kev ua yeeb tshuaj ua raws li qhov tseem ceeb, nws tuaj yeem ua rau kom muaj kev cuam tshuam cov tshuaj - metabolizing enzymes thiab thauj. Muaj zog incers tuaj yeem txhawb ntau ntau cov noob ua ke, xws li PheyBarbital Induction of CYP2B6, CYP2C9, CYPT, thiab ntau thauj cov protein zoo li mrp2. Thib ob, muaj cov tsiaj txhu - tshwj xeeb sib txawv hauv qhov hepatocytes teb rau cov yuam sij. Piv txwv li, Rifampicin yog ib qho txiaj ntsig zoo rau tib neeg thiab luav Hepatocytes, tab sis tsis muaj kev cuam tshuam nrog kev cuam tshuam ntawm cov hepatocytes. Thaum kawg, suboptimal plating ntuitity ntawm platable hepatocytes yuav ua rau txo cov lus qhia ua ntej ntawm P450s thiab artificion ordreed inductes teb. Raws li pom hauv daim duab 3, Hepatocytes ntawm qis dua plating ua yeeb yam pom kev ua si basal ntawm CYP1A2, CYP2B6, thiab CYP3A4, tab sis CYP3A4, tab sis CYP3A4, tab sis muaj zog induction teb. Yog li no, Hepatocytes noj qab nyob zoo ntawm qhov kev ua haujlwm sib txawv yog qhov tsim nyog los tau txais physiologically cuam tshuam.

Daim duab 2. Kev sib raug zoo ntawm plating ntom ntawm crypreserved tib neeg hepatocytes thiab enzyme induction

Source: Tam sim no kev tshawb tshuaj tshuaj tam sim no thev naus laus zis, 2010, 7: 188 - 198

2). Kev ntsuas Hepatotoxicity: Saib xyuas morphological nyob rau hauv lub qhov tsom tsom, xws li cell morphohology, vacpet thiab lipid droupet aggration, thiab cell txuas / detachment. Nrhiav tau ntawm Hepatosocyte Necrosocyte (raws li tau hais los ntawm aspartate aminottransferase, thiab alopine aminottransferase) thiab agoptosis (DNA tawg). Rau cytokine - Kho Cytotoxicity, Ib Leeg Monolayer Cov kab lis kev cai tsis tuaj yeem kwv yees cov lus teb tsis tau los ntawm cov kev cai tsis txaus siab rau cov neeg nyob sib raug zoo - parenchymal hlwb, thiab sinusoidal endothelial lub hlwb.

3). "Relay Txoj Kev" rau kev kawm txog qeeb metabolizing compound clearance thiab nws cov metabolites: Cov kev ua yeeb yam tshuaj yeeb - metabolizing enzymes hauv platable hepatocytes pib txo qis tom qab 24 teev ntawm plating. Tom qab ua kom lub platable hepatocytes nrog rau ntshav qab zib - Muaj nruab nrab ntawm cov kev txaus siab tau 24 teev, tom qab ntawd hloov mus rau cov lus qhia yooj yim Hepatocytes rau kev kawm ntxiv (Daim Duab 3).

Daim duab 3. Platable Hepatocyte Relay Txuj Kev Hloov Cov Txheej Txheem

Source: Cov tsiaj ntawv metabolism, 2016, 10: 3 - 15

3). Txheeb xyuas lub cellular uptake, Endocyttosis, Endosomal khiav tawm, thiab cov teebmeem ua si muag ntawm lub hom phiaj ntawm Hepatocyte - Lub hom tshuaj acid me me kua qaub.

Sandwich sau qauv qauv

Nyob rau hauv vitro, Hepatocytes tuaj yeem yog kev coj noj coj ua ntawm ob txheej ntawm collagen lossis matrigel kom rov tsim dua hauv cov qauv Vivo, hu ua Sandwich sau. Hepatocytes kev coj noj coj ua ntawm ob txheej ntawm gel - collagen (sandwich qauv) tuaj yeem txhim kho cov qauv morphology thiab kev muaj peev xwm txhim kho cov hlwb, thiab tswj hwm lawv cov haujlwm rau lub sijhawm ntev dua. Tsis tas li ntawd, Hepatocytes hauv Sandwich Culture tuaj yeem kho kom tau cov basolateral thiab canalicular thauj neeg cov khoom siv bile (Daim Duab 4).

Daim duab 4. Cov lus qhia ntawm cov neeg thauj khoom hauv tib neeg Hepatocyte Sandwich Qauv

Source: Tam sim no kev tshawb tshuaj tshuaj tam sim no thev naus laus zis, 2010, 7, 188 - 198

Cov Ntawv Thov:

1. 1). Kwv yees ntawm biliary licretion ntawm cov tebchaw.
2. 2). Kev ntsuam xyuas cov hepatic thiab biliary faib ntawm endogenous thiab exogenous sib txuas thiab cov metabolites.
3. 3). Kev kwv yees kev kho kho kom meej los ntawm cov metabolism thiab thauj khoom, thiab tsim kev ua haujlwm ntawm lub cev - Raws li cov kws ua haujlwm.
4. 4). Kawm Hepatotoxicity thiab muab cov tshuab siv rau tshuaj tivthaiv tshuaj - inducts raug mob. Cov ntaub ntawv tau muab cov ntaub ntawv ua ke ua ke los ua cov qauv chaw muag tshuaj kom kwv yees tshuaj muaj peev xwm ua rau tib neeg lub siab.
5.  
   1. 3. Thawj Hepatocyte 3D Kab lis kev cai

6. Hauv cov kab lis kev cai 3D kev cai, Hepatocytes yog kev coj ua hauv ib peb - Qhov av loj, uas zoo dua cov lus qhia hauv vivo cov nplooj siab tau muab piv rau 2D monolayer haiv neeg. Cov kab ke no txhawb nqa ntawm tes - Cell thiab cell - kev sib cuam tshuam ntawm daim siab ua haujlwm, suav nrog cov kev ua haujlwm yeeb tshuaj, suav nrog cov tshuaj khib nyiab, cov protein tso tawm, thiab cov kua tsib. Thawj Hepatocyte 3D kev cai yuav siv tau los kawm lub siab - cov kev ua tau zoo rau kev kuaj tshuaj, kev ntsuas tshuaj lom, thiab cov qauv kab mob.

   Kev Ua Qauv Zoo

Los ntawm cov tswv yim xws li ultra - Cov kab lis kev cai txuas nrog, thiab cov kab lis kev cai xov xwm cell (Daim Duab Hepatocitate tuaj yeem sib sau ua ke nrog 150 lub cheeb tsam tsis muaj kev vam khom rau sab nraud matrices. Ib qho zoo dua ntawm kev coj noj coj ua yog tias txhua lub spheroid tsuas yog 1,330 - 2,000 lub hlwb, txo cov hlwb piv rau lwm yam kev coj ua 3D. Cov kev tshawb fawb tsis ntev los no tau pom tias thawj coj noj coj ua yeeb yam zoo li 5 lub lis piam, nrog rau kev hloov pauv ntawm cov tshuaj, thiab ua haujlwm zoo dua rau 14 hnub hauv kev coj noj coj ua. Txawm li cas los xij, daim siab yog ntau dua cov complex tshaj ib cell ua ke. Cov basodateral sab ntawm Hepatocytes cuam tshuam nrog cov ntshav, uas yog qhov tseem ceeb ntawm lub cev ua haujlwm sib dhos, thiab qhov no tseem tsis tau rov qab ua qauv.

Cov Ntawv Thov:

1. 1). Kev kawm ntawm qeeb metabolizing compound clearance thiab nws cov metabolites.
2. 2). Hepatotoxicity Tshawb Fawb.
3. 3). Kev ntsuam xyuas ntawm cellular uptake, Endocytosis, endosomal khiav tawm, thiab cov kev sib tw ntsiag to ntawm cov hom phiaj me me kua qaub kua txiv.

Daim duab 5. SpHeroid Kev Lag Luam Txuj Ci

Qhuab Orgoids Qauv

Cov kev pom zoo ntawm cov kabmob ntawm cov kabmob, muaj peev xwm ua kom muaj zog hauv Vivo Microenterviror thiab Cell - rau - Kev sib cuam tshuam ntawm tes. Cov kev sim siab tau pom tias yog tus qauv tshaj plaws rau tib neeg lub siab tshawb fawb txog kev tshawb fawb.

Kev tsim kho ntawm tes rau kev sim ua kom haum:

① Pluripotent Qia Hlua (PSCS):

Emigrya ​​tso qia (escs) thiab induced pluripotent qia hlwb (IPSCS) muaj peev xwm siab pluripotency, muaj peev xwm unlimited proliferative muaj peev xwm. Nyob rau hauv cov cawv ntawm cov paib tshwj xeeb, lawv sib txawv rau hauv Hepatocyte - Zoo li cov hauj lwm nrog kev ua si thiab muaj nuj nqi. Txawm li cas los xij, daim siab ua haujlwm tau muab los ntawm PSCS tuaj yeem dhau epigenetic thiab cov kev hloov caj ces, nthuav qhia cov kev hloov pauv tsis zoo rau lub sijhawm ua kom tawg.

Cov Ntawv Thov:

1. 1). Genetic Liiv Kab Mob Kab Mob
2. 2). Kis kis tus kab mob qauv
3. 3). Kev Ntsuas Tshuaj CyTotoxicity

② daimem ntxig - Cov hlwb los ntawm cov hlwb: cov no suav nrog cholangiocytes thiab hepatocytes. Paub tab Hepatocytes khaws cov txiaj ntsig qia ntawm lub peev xwm thiab muaj peev xwm proliferative hauv ib qho chaw tshwj xeeb. Piv rau psc - Cov Ecods, Orgived los ntawm thawj cov nqaij mos, thiab tswj cov penotypic ntau dua, thiab kev ruaj khov nyob rau hauv ntev - sij hawm hauv vitro kab lis kev cai. Txawm li cas los xij, lub sijhawm ntev - Lub sij hawm prolderative muaj peev xwm ntawm mature Tib neeg Hepatocyte piv rau cov me nyuam hepatocytes lossis cov laus gete hepatocytes. Kev coj noj coj ua tus neeg laus hepatocyte orgernoids tseem nyuaj.

Kab lis kev cai Txuj Ci (Daim Duab 6): Daim Ntawv Mob Ntsej Muag yog zom rau ib lub hlwb, thiab cov phaj zoo nkauj kom tsim dome - qauv qauv. Txo nyob rau hauv ib lub cell kev cai tswj hwm (37 ° C) li 15 feeb. Tom qab solidification, ntxiv cov kab lis kev cai me me. Zaj tom qab kwv yees li 14 hnub. Hloov cov nruab nrab nrog txawv nruab nrab tom qab 7 - 10 hnub.

Cov Ntawv Thov:

1. 1). Hepatotoxicity Qauv
2. 2). Nyob rau hauv vitro metabolism tsis sib haum kev kawm
3. 3). Tsis - Cov Kab Mob Khov Rog Rog Rog
4. 4). Kev Tsim Tshuaj Tshuaj rau Kev Mob Siab Benign thiab Malignant Kab Mob

Daim duab 6. Kab lis kev cai thiab cov txheej txheem tso cai ntawm cov ntaub so ntswg - Tau Txais Kev Mob Siab

Source: Cell & Bioscience (2023) 13: 197

Kev sib piv ntawm kev coj noj coj ua thiab kev coj noj coj ua mob

4. Thawj Hepatocyte Co - Kab lis Qauv Qauv

1. 1. 2D Qhov Tseem Ceeb Hepatocyte Co - Kab lis Qauv Qauv
      

      Hauv 2D thawj Hepatocyte CO - Cov qauv kev coj ua, ob lossis ntau dua cov xov tooj ntawm tes yog sib xyaw thiab kev coj noj coj ua hauv ob qho chaw -. Cov ntsiab lus tseem ceeb ntawm cov qauv no yog kev sib txuam ncaj qha ntawm cov xov tooj ntawm cov xov tooj ntawm cov cytokines thiab kev sib txuas lus tsis sib xws thiab tshuaj sib txuas lus. Thawj Hepatocyte Functions, xws li albumin ntau lawm thiab tshuaj metabolism muaj peev xwm, tuaj yeem tswj tau ntev txog peb lub lis piam.

      Cov Ntawv Thov:

      1. 1). Thawj Hepatocytes CO - Kev coj ua nrog fibroblasts: Tus qauv no yog siv rau kev kawm cov kev tshem tawm qeeb qeeb - metabolizing sib txuas thiab lawv cov metabolites.
      2. 2). Thawj Hepatocytes CO - Cultured nrog tsis - Parenchymal Nplooj Siab, vim nws yuav pab tau daim siab ua tshuaj ntsuab (Dili), raws li nws tau cog lus rau daim siab
      3. 3). Thawj Hepatocytes CO - Kev coj noj coj los yog siv los tshawb xyuas cov kev siab ntsws tshuaj yeeb - Lus qhia cell teb.
      4. 4). IPhase's hepatomax ™Co - kab lis kev cai teeb meem: Cov Ipase tau tsim kho cov co - kab lis kev cai nrog thawj Hepatocytes los ntawm ntau hom, hu ua Hepatomax™Cov. Los ntawm cov - Kev coj noj coj ua tib neeg Hepatocytes nrog cov tshuaj stromal, nws yog qhov ua tau kom tswj cov tshuaj zoo - metabolizing enzyme kev ua hauv tib neeg hepatocytes kom txog 3 lub lis piam. Qhov system no haum rau kev kawm qeeb - metabolizing cast clearance tus nqi thiab lawv cov metabolites.

      Cov qauv sib xyaw no yog suav nrog cov khoom siv hluav taws xob ntau ntxiv, tshuaj lom neeg, thiab kev saib xyuas kab mob sib txawv, kev pom zoo li cas ntawm tes ua rau mob siab ua haujlwm thiab kab mob.
      
      

      3D Qhov Tseem Ceeb Hepatocyte Co - Kab lis Qauv Qauv

      Direct 3D co - Kab lis kev cai: Tus qauv no cuam tshuam ob lossis ntau hom kev sib tw ntawm lub hlwb (EPLYOIDAL Direct 3D Coache - Kev Noj Tswm Muaj Tso Cai Zoo Sib Tsoo ntawm cov tshuab sib txawv ntawm soluble cytokines, thiab extracellular mytoxines, thiab kev sib txuas lus ntawm hepatocytes.

      Cov Ntawv Thov:

      1. 1). Daim siab ua haujlwm fibrosiscosis: siv los kawm cov txheej txheem thiab kev nce qib ntawm lub siab fibrosis.
      2. 2). Tshuaj - Raug Mob Mob Siab (Dili) Qauv: Pab simulate thiab ntsuas lub siab puas tsuaj los ntawm cov tshuaj.
      3. 3). Kev sib cuam tshuam yeeb tshuaj, tshuaj yeeb tshuaj yeeb, thiab siv cov tshuaj enteruction li cas, lawv yuav ua li cas sib txawv, thiab lawv ua li cas rau lub siab ua kom tawv nqaij.

      Indirect 3D co - Kab lis kev cai: Cov qauv no siv lub cev sib cais (piv txwv li, transwell lossis ntau hom kev coj noj coj ua) rau ntawm 3D ib puag ncig (chaw ncaj qha Cov kev sib txuas lus nruab nrab ntawm cov cells tshwm sim ntawm soluble cytokines.

      Cov Ntawv Thov:
      Siv rau kev kawm cov lus tsis - sib tham ntawm cov hauj lwm cov hauj lwm hauv lub cev.

      
      Hauv cov ntsiab lus, cov ntsiab lus qub, ua ib tus thawj coj hauv cov kev tshawb fawb ntxiv roj ntsha, muab cov kev daws teeb meem tshuaj rau kev kuaj tshuaj tsis raug tshuaj kho. Los ntawm kev cais thiab kev coj noj coj ua ntawm cov thawj coj hauv ntau hom rau kev txhim kho cov ntaub ntawv txhawb nqa - Cov cuab yeej tshawb fawb txog kev tshawb fawb vitro tshawb fawb thiab kev loj hlob. Peb yog ib tus khub uas ntseeg siab rau cov neeg siv khoom hauv kev lag luam kws tshuaj, tau cog lus los muab kev daws teeb meem ntug rau cov kev daws teeb meem tsis yog soj ntsuam.