Yekutanga Hepatocytes: Chishandiso chakakosha kufambira mberi zvisiri 1

Yekutanga Hepatocytes: Chishandiso chakakosha kufambira mberi zvisiri 1

Chiropa, seyemhando yekutanga yekufumurwa nemishonga yezvinodhaka, inotora yakakosha basa mune zvinodhaka metabolism uye zvinhu zvine chepfu. Primary hepatocytes yerefu yeCelluly hunhu uye mazano emhando ye enzymes naCytochercome p450 Nechikonzero ichi, yekutanga hepatocyt inoonekwa zvakanyanya segoridhe yakagadzirwa muVitro chombo mhando uye inofarirwa nevaongorori mukudyidzana, zvinodhaka metabolism, uye zvinhu zvepfuma. Chinyorwa ichi chinopa kutarisisa kwe 2d uye e3D tsika mhando zvichienderana nePrimary hepatocytes uye zvavanoshandisa mukuvandudzwa kwezvinodhaka.

Keywords: Primary hepatocytes, 2D kurimwa, 3D kuridzwa, magadzirirwo, CO - Tsika.

1. 1. Kuzviparadzaniswa kwePrimary hepatocytes

Kuzviparadzanisa kwekutanga kwehipatocytes inguva yakaoma mukutanga muVitrolocha Models, neyairi - Nhanho yeColagenase Perfusion Nzira Kuve Akanyanya kushandiswa. Muzvipfuyo zvidiki, chiropa chinowanikwa kuburikidza neiyo portal vein kana yakaderera vena cava, nepo mhuka hombe kazhinji dzinoda kutsvaira kuburikidza nechiratidzo chemoyo kana zvikamu. Zvinhu zvinoverengeka zvinokonzeresa zvinopesvedzera kuzviparadzaniswa kwehepatocytes: kutanga, collagenase inofanira kunge isiri - cytotoxic. Chechipiri, nguva yekugaya ndeyechokwadi-Vese vari pasi - kugaya uye kugadzirira - kugaya kunogona kukanganisa hepatocyte goho uye kusanganisa. Chechitatu, chirwere chechiropa chinofanira kuve chakakwana, sehematecytes zvakanyanyisa kutarisana neIschemic kukuvara. Chiropa chinoshandiswa kugadzirira hepatocyte chinofanira kunge chakakurumidza kutonhorera kuti uderedze metabolic mitengo uye kudzivirira metabolic hypoxia uye inotevera ischemia.

Zviyero zvePrimary Hepatocytes zvinoshandiswa mukutsvaga zvinodhaka ndezvekutevera:

1. 2. Primary hepatocyte 2d tsika

Kumiswa kweHepatocyte modhi

Iyo ine zvinodhaka zvakazara - metabolizing enzymes uye coffeks, ichiita kuti iite yakakodzera kudzidza akasiyana metabolic clearance nzira. Nekudaro, iyo yekuvhara yekumiswa hepatocytes uye chiitiko chezvinodhaka - Iyi modhi inowanzo shandiswa kufungidzira kubvumidzwa kwezvinodhaka zvine mwero kune yakanyanya kujekesa mitengo. Kana iyo clearance rate isingasviki 20%, tsika dzakarurama clearance hazvigone kutsanangurwa. Kuparadzaniswa kwechinyakare Hepatocyte - Yakavakirwa muVitro Metabolism Models haina kukwana kuti igadzire (metabolizing makomputa avo kufanotaura marongero ekujekesa uye metabolic zvigadzirwa zveaya makomendi. Kumiswa kweHepatocyte Rekutamba nzira (Mufananidzo 1) inogona kushandiswa kuwedzera iyo incube nguva kusvika maawa makumi maviri kana kunyange kureba.

Chikumbiro:Enzyme chiitiko uye metabolic kugadzikana zvidzidzo zve madhiri madiki madhiragi.

Mufananidzo 1

Kunobva: Metab Metab Unhivha, 2012,40 (9): 1860-1865

Iyo inonzwisisika hepatocytes modhi

Primary hepatocytero mune imwe 2D system paColagenter - Yakachinja Tsika Mahwendefa. Iyo hepatocytes inoratidza epithelial morphology, ine protruduring nuclei, kazhinji ichiratidzira muchimiro chakarukwa. Izvo zvinokwanisika zveimwe hepatocyte bonolayer tsika inosanganisira: 1. Kuenderera kwesero polarity uye basa.2. Kushaikwa kwemamwe marudzi akakodzera echitokisi (i.e., non - Pangerchymar masero) ayo anodikanwa kuti iite basa rakajairika.3. Kukundikana kupa zvakaringana zvinovaka muviri uye kupedzisa parcine kutsigira iyo hepatocytes mukuita mabasa avo (senge bile acid uye serum protein biosynthesis).

Zvishandiso:

1). Kuongorora kwezvinodhaka - Kubata kwezvinodhaka:Izvi zvinosanganisira enzyme induction, enzyme inhibition, uye zvekutakura zvidzidzo. Chekutanga, kana paine zvinodhaka sezvakaita sidocer, inogona kutungamira pakuratidzira kwezvinodhaka - metabolizing enzymes uye vatakuri. Instrucrs yakasimba inogona kukwikwidza marudzi akawanda panguva imwe chete, yakadai sechiitiko chekunakirwa kweCyp2b6, Cyp3a4, Cyp2c9, Ugp, uye akati wandei mapuroteni anotakura MRP2. Kechipiri, kune mhuka dzakasiyana - Kusiyana kwakananga mumabatirwo aHepativecytes vanopindura kune vanokurudzira. Semuenzaniso, rifampicin inyanzvi inoshanda yemunhu uye rabit hepatocytes, asi haina indiction mhedzisiro pane rat hepatocytes. Pakupedzisira, suboptimal plating density yeanonetesa hepatocytes inogona kutungamira kuderedzwa kweiyo basal kutaura kweP450s uye yakawedzera kuwedzera mhinduro. Sezvinoratidzwa mumufananidzo 3, hepatocytes pazasi pedenties inoratidza yakadzika basal basa reCyp1, cyp2b6, uye cyp3a4, asi mhinduro dzesimba rekutsamwa. Naizvozvo, hutano hepatocytes dzakakodzera planity dzakakosha kuti uwane data phepfeldiology yakakodzera.

Mufananidzo 2. Ukama pakati pekuisa density yeClyypress huwandu hweHepatocytes uye enzyme induction

Kunobva: Yazvino Dhivha Discovery Technologies, 2010, 7: 188 - 198

2). Hepatototoxity Ongororo. Kuonekwa kweHepatocyte necrosis (sezvakaratidzwa neAppartate aminotransfease, lactate dehydrogease, uye alanine Aminotransfase) uye Apoptosis (DNA Sraugmentation). For Cytokine - Singleated Cytotoyer tsika, tsika dzepamusoro dzisingakwanisi kufanotaura nemirairo ine mitezo

3). "REALY NYAYA" yekufunda yekudzora zvishoma metabolizing canpound clearance uye yayo metabolites. Mushure mekusimudzira anongwarira hepatocytes neSerum - Mahara epakati ane iyo yekunyepedzera kwemaawa makumi maviri nemana, iyo svikiro, yakabva yatamiswa, yakabva yasanganiswa.

Mufananidzo 3. Anotya Hepatocyte anodzokorora nzira yekufambisa maitiro

Kunobva: Zvinodhaka metabolism mavara, 2016, 10: 3 - 15

3). Ongorora iyo Cellular Bokal

Sandwich Chiravira

Mu vitro, hepatocytes inogona kuvezwa pakati pezvikamu zviviri zvecolagen kana Matrigel kudzoreredza muVivo Storures, inozivikanwa seSandwich kuridzwa. Hepatocytes tsika pakati pezvikamu zviviri zveGEL - Colagen (Sandwich (Sandwich) inogona kuvandudza morphology uye kusarudzika kwemasero, uye chengetedza mashandiro avo kwenguva yakareba. Kupfuurirazve, hepatocytes mune sandwich tsika inogona kudzoreredza porlarity, kubvumira mamiriro ezvekutakurwa kwe basoisteral uye canalicular vatakuri, pamwe nekuumbwa kwekushanda bile duct network (Mufananidzo 4).

Mufananidzo 4

Kunobva: Yazvino Dhivha Discovery Technologies, 2010, 7, 188 - 198 - 198 -

Zvishandiso:

1. 1). Kufungidzira kushamisika kwehuwandu hwemakomisheni.
2. 2). Kuongorora iyo Hepatic uye biliary kugoverwa kweiyo endofius uye neoxanhuus makomputa uye metabolites.
3. 3). Fungidzira kujekesa kujekerwa ne metabolism uye vatakuri, uye kugadzira physiology - based pharmacokinetic mhando.
4. 4). Kudzidza hepatoxity uye kupa michina yemishonga yekiriniki yemishonga - yakakuvadzwa chiropa kukuvara. Dhata inobatanidzwa mune systems pharmacology mhando kuti ufanane zvinokwanisika zvinodhaka - kukurudzira chiropa muvanhu.
5.  
   1. 3. Primary Hepatocyte 3D tsika

6. MuC 3D Tsika Systems, Hepatocytes mune matatu (Dimensional Matrix, ndeipi iri nani kutevedzera iyo muVivo yeVivo yeVivolocha yekugadzira yakafananidzwa ne 2D monolayer tsika tsika. Iyi masisitimu inosimudzira cell - cell Yepamusoro Hepatocyte 3D tsika inogona kushandiswa kudzidza chiropa

   Spheroid Model

Kuburikidza nehunyanzvi sekuti Ultra - Yakadzika yekubatanidza Tsika, uye Yakaturika Cell Cell Imwe mukana wetsika yakasanganiswa ndeyekuti imwe neimwe spheroid inoda chete 1,3 - Zviuru zviviri - Zviuru zviviri kudzora huwandu hwemasero uchienzaniswa nedzimwe nzira dze3D tsika. Zvidzidzo zvichangobva kuitika zvakaratidza kuti hepative tsika dzepfungwa dzinogona kuraramisa kwemavhiki anosvika mashanu, kufananidzwa kweshoko rekutora kwezvinodhaka. Nekudaro, chiropa chakanyanya kuoma pane cell acgregate. Iyo basolateraleral rutivi rwe hepatocytes inobata neropa, nepo bile inoyerera kubva parutivi rwepapic, ichi chinhu chakakosha chechimiro chakaomarara, uye izvi hazvigone kudzokororwa mumhando yeiyo spheroid.

Zvishandiso:

1. 1). Kudzidza kweanononoka metabolizing canpound clearance uye yayo metabolites.
2. 2). Hepatotoxity kutsvagisa.
3. 3). Kuongororwa kweCellular Bokal, Endocytosis, endosomal kupukunyuka, uye kupedza kupukunyuka pane chinangwa majenji eHepatocyte - Targeted madiki madiki mishonga yemishonga.

Mufananidzo 5. Spheroid tsika nzira

Chiropa organiids modhi

Tsanangudzo yekubvumirana yezviratidzo: 3D zvivakwa zvinobva pamasero ematombo, kana masero akasarudzika, anokwanisa kudzvanywa microdentement uye cell - kuti - Chiropa magadzirirwo akaonekwa semhando yepamusoro yepamusoro yeropa remunhu rekutsvaga biology kutsvagira.

Masero emuchina weropa organia

① PluripoTent STEM masero (PSCS):

Embryonic Stem masero (ESCS) uye akakwenenzverwa pluripotent STEM masero (IPSCS) ane yakakwirira plugupotency, planticity, uye isina mugari, uye isina muganho wekuwedzera. Pasi pesimba rezvakananga zvinhu zvemainirimo, ivo vakasiyanisa muHepatocyte - Semasero ane chekuita nebasa. Nekudaro, zvirongo zvechiropa zvinotorwa kubva kuPSCS zvinogona kuve neEpigenetic uye genetic shanduko, kuratidzira chromoumal aeuploidy shanduko panguva yekuwedzera.

Zvishandiso:

1. 1). Genetic chiropa chirwere chemhando
2. 2). Zvinotapukira chiropa mhando mhando
3. 3). Zvinodhaka Cytoticity Kuedza

② ROPA Tishu - Yakatorwa Masero: Izvi zvinosanganisira cholangiocytes uye hepatocytes. Mature hepatocytes chengetedza stem cell cell inogona uye yekuwedzera kugona mune yakatarwa nzvimbo. Kuenzaniswa nePSC - Yakatorwa magadzirirwo, magadzirirwo anobva kuTisuyes Achikura, ane manyorerwo akagadzikana, uye chengetedza kugadzikana uye kugadzikana kweiyo vitro tsika. Nekudaro, iyo yakareba - term probliferative kugona kwemunhu akakura vanhu Hepatocyte emunhu ane mashoma achienzaniswa neFetal Humitown Hepative Hepative Hepatows. Tsika Yemunhu Adult Hepatocyte zvigadziriso zvinoramba zvakaoma.

Tsika nzira (Mufananidzo 6): Chiropa tishu chakachekwa mumasero mumwe, uye musanganiswa waMatrigel uye masero akareba musi wa 24 - zvakanaka ndiro yekuumba dome - zvivakwa zvakaumbwa. Incubate mune imwe cell tsika incubator (37 ° C) kwemaminetsi gumi nemashanu. Mushure mekusimbisa, wedzera chaiyo tsika yepakati. Ndima mushure memazuva gumi nemana. Tsiva iyo yepakutanga yepakati neyakasiyanisa yepakati mushure me7 - Mazuva gumi.

Zvishandiso:

1. 1). Hepatotoxity Models
2. 2). Mu vitro metabolism disders
3. 3). Kwete - chidhakwa nehasha
4. 4). Budiriro yezvinodhaka yeBenignant neakashata chirwere chirwere

Mufananidzo 6. Tsika uye ndima yekuita tishu - inotorwa miganhu yeropa

Kunobva: Cell & Bioscience (2023) 13: 197

Kuenzanisa kwetsika yakasviba uye tsika orgooid

4. Primary Hepatocyte CO - Tsika Yemuenzaniso

1. 1. 2D Primary Hepatocyte CO - Tsika Yemuenzaniso
      

      Mune 2D Primarcy Hepatocyte CO - Tsika yemhando, maviri kana anopfuura akasiyana masero marudzi akasanganiswa uye ane tsika mune maviri - nzvimbo dzakaderera. Chinhu chakakosha chekutaurirana kwakananga pakati pemhando dzakasiyana dzemhando dzeSero, kuwirirana pakati pemasero uye yekuwedzera matrix, kana iyo isina kuchinjika chiratidzo kuburikidza neCytokines uye Cytokical uye Cytokical uye Cytokical uye Cytokical uye Cytokical Cytokusines uye makemikari ekukurukurirana. Basa rekutanga hepatocyte, senge albumin kugadzirwa uye zvinodhaka metabolism kugona, kunogona kuchengetedzwa kwemavhiki matatu.

      Zvishandiso:

      1. 1). Primary HePatocytes CO - Yemhando yeFibroblasts: Iyi mhando inoshandiswa kudzidza kuongororwa kwekujeka kwekunonoka - metabolizing makomputa uye metabolites yavo.
      2. 2). Spent Hepatocytes CO - YEMAHARA YEMAHARA YEMAHARA CELS (I.COM INOGONESESWA KUTI SIMBA, CHEMOKINES SHOKO REMUFANANIDZO WEMAHARA.
      3. 3). Primary HePatocytes CO - YEMAHARA AT CELS: Iyi mhando inoshandiswa kuona livers zvinodhaka metabolism - yakatarwa t masero mhinduro.
      4. 4). Hepase's Hepatomax ™CO - Tsika Yetsika: Iphase yakagadzira CO CO - Tsika Tsika Ine Primary Hepatocytes kubva kune dzakasiyana mhando, inozivikanwa seHepatomax™. NaCo - Tsika Hunhu Primary Hepatocytes ine Stromatowner masero, zvinokwanisika kuchengetedza zvinodhaka - metabolizing enzyme chiitiko muHepatocytes inosvika kumavhiki matatu. Iyi sistimu yakakodzera kudzidza inononoka - metabolizing yakamanikidzwa clearance metate uye yavo metabolites.

      Uyu CO - Tsika Yemhando inopa zvakanyanya phepfeligigicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicicical yedhisheni, uye chiropa -
      
      

      3d Primary Hepatocyte CO - Tsika Yemuenzaniso

      Direct 3d CO - Tsika: Iyi muenzaniso inosanganisira kusanganisa maviri kana anopfuura akasiyana masero emuchato Direct 3D Co CO - Tsika Inobvumira Kubatana kwepedyo pakati pechiropa chakasiyana kuburikidza neCell Cytokines.

      Zvishandiso:

      1. 1). Chiropa fibrosis modhi: yakashandiswa kudzidza iyo michina uye kufambira mberi kwechiropa fibrosis.
      2. 2). Mushonga - Yakamisikidzwa Kukuvara Kwechiropa (Dili) Model: inobatsira kutevedzera uye kuongorora chirwere chekukuvara kunokonzerwa nemishonga.
      3. 3). Kubata kwezvinodhaka, zvinodhaka metabolism, uye enzyme induction: inoongorora kuti akasiyana mishonga inobatana, sei metabolized, uye kuti vanonyengera sei miropa enzymes.

      Indiirect 3D Co - Tsika. Kukurukurirana pakati pemasero kunoitika kuburikidza nekunyaradza cytokines.

      Zvishandiso:
      Inoshandiswa pakudzidza isiri - Bata kutaurirana pakati pechiropa chemutumbi mumuviri.

      
      Mukupfupikisa, Iphase, semutungamiri muVitro Biological kutsvagudza, inopa mhinduro dzakakwana dzeNonosvika - kuongororwa kwekiriniki zvinodhaka. Kubva pakuzviparadzanisa uye tsika dzekutanga hepatocytes kubva kune dzakasiyana mhando kune zvigadzirwa zvekutsigira senge manyorerwo ezvakanyanya kuita zvekushandisa mukuwanikwa kwekutsvaga kweVitro uye kusimudzira. Isu tiri munhu anovimbwa naye kune vatengi muindasitiri yemishonga, akazvipira kupa kucheka - EGED Solutions yeNo isiri - Clinical tsvarhurumende.