Chiwonetsero chachikulu: chida chofunikira kwambiri popititsa patsogolo Not - Chachipatala mu kafukufuku wa Vitro

Chiwonetsero chachikulu: chida chofunikira kwambiri popititsa patsogolo Not - Chachipatala mu kafukufuku wa Vitro

Chiwindi, monga chiwalo choyambirira cha kuwonekera mankhwala, kumatenga mbali yofunika kwambiri pa kagayidwe ka mankhwala ndi kafukufuku. Makina oyambira a hepatocytes omwe ali ndi ma cell a ma cell ndi matope, kuphatikiza membrane - Kusakaniza netoprome P450 Pachifukwa ichi, hepatocytes yoyamba imawonedwa ngati muyeso wa golide wopangidwa mu Vitro chiwindi ndipo amakondedwa ndi ofufuza pogwiritsa ntchito mankhwala osokoneza bongo, kagayidwe mankhwala. Nkhaniyi imapereka mwachidule mitundu ya 2d ndi 3D yochokera ku hepatocyte yoyamba ya hepatocyte ndi mapulogalamu awo popanga mankhwala.

Mawu osakira: Hepatocytes woyamba, kulima 2D, kulima 3 - zinthu, zinthu zina, chikhalidwe.

1. 1. Kupatula kwa hepatocyte

Kupatula kwa hepatocytes koyambirira kwa hepatocytes ndi gawo lovuta kukhazikitsa mitundu ya vitro, ndi awiriwo - Pulogalamu iwiri - Mu nyama zazing'ono, chotupa cha chiwindi chitha kuchitidwa kudzera mu vein kapena kutsika kwa vena, pomwe nyama zazikulu zambiri zimafunikira zonunkhira kudzera pa chiwindi kapena magawo. Zinthu zingapo zazikulu zimapangitsa kuti zikhale zowoneka bwino za hepatocytes: Choyamba, collagenase ziyenera kukhala - cytotoxic. Chachiwiri, nthawi ya chimbudzi ndi yofunika-Onse ochepera - chimbudzi ndi zopitilira - chimbudzi chitha kuletsa zipatso za hepatocyte ndi mavuto. Chachitatu, chiwindi chizikhala choyenera, monga hepatocytes amakhudzidwa kwambiri ndi kuwonongeka kwa ischemic. Chiwindi chomwe chimagwiritsidwa ntchito pokonzekera hepatocyte chizikhala cholumikizidwa mwachangu kuti muchepetse misonkhano ya metabolic komanso kupewa kagayidwe kachakudya ndi Ischemia.

Miyezo ya hepatocytes yoyambirira yogwiritsidwa ntchito pofufuza mankhwala ndi motere:

1. 2. Hepatocyte 2D

Chiwonetsero cha Hepatocyte

Muli ndi mankhwala okwanira - michere yopukusira ndi matongosolo, kupangitsa kukhala koyenera kuphunzira njira zingapo za metabolic. Komabe, kusokonekera kwa hepatocyte ndi ntchito ya mankhwala - Mikangano yopumira imachepa kwambiri ngati nthawi ya makulitsi kumawonjezeka kwa maola 4. Mtunduwu umagwiritsidwa ntchito kuwerengera mankhwala osokoneza bongo omwe ali ndi zochulukirapo mpaka ziphuphu zapamwamba. Pamene chilolezo choledlerance chili ndi 20%, mfundo zolondola zolondola sizingatsimikizidwe. Kuyimitsidwa kwachikhalidwe hepatoctte - Kutengera mtundu wa Vicro metabolism ndi kusakwanira kwa kapangidwe ka kachulukidwe kakang'ono ka kagayidwe ka kagayidwe ka kachakudya. Njira yoyimitsidwa ya hepatocyte yolumikizira (Chithunzi 1) itha kugwiritsidwa ntchito kukulitsa nthawi ya makulitsidwe mpaka maola 20 kapena kuposerapo.

Ntchito:Ntchito ya enzyme ndi statice yokhazikika ya mankhwala osokoneza bongo azolekyulu.

Chithunzi 1. Kuyimitsidwa Hepatocyte Gouty njira yosamutsa

Source: Ma Metable adatumiza mankhwala, 2012,40 (9): 1860-1865

Chitsanzo cha Hepatocytes

A HOPACECYTE yoyambirira ya hepatocytes imayamikiridwa mu dongosolo la 2d pa Collagen - Chikhalidwe Chachikhalidwe cha Chikhalidwe. Ma hepatocytes amawonetsa ma epitelial morphology, omwe ali ndi nucleing nuclei, nthawi zambiri amapezeka mu mawonekedwe a sayansi. Zovuta za Chikhalidwe chokhacho cha hepalayerte monolayerte cololayerte ndi: 1. Kusintha kwa cell polartity ndi ntchito.2. Kuperewera kwa mitundu ina yam'manja (i.e., No - maselo opezeka pazenera) omwe amafunikira kuti azigwira ntchito.3. Kulephera kupereka michere yokwanira ndi zinthu zothandizira paracy kuti zithandizire hepatocytes popanga ntchito zawo (monga bile acid ndi serum protein biosynthesis).

Mapulogalamu:

1). Kuyika Mankhwala - Kugwirizana ndi Mankhwala:Izi zikuphatikiza enzyme ikuluikulu, enzyme zoletsa, ndi kafukufuku wonyamula. Choyamba, pamene mankhwala amagwira ngati chochititsa chidwi, zitha kubweretsa kufotokozera kwa mankhwala - michere yosungunuka ndi zonyamula. Okakamiza mwamphamvu amatha kukweza majini angapo nthawi imodzi, monga phenobarbital incution ya cyp2b6, cyp3a4, cyp3c9, ugt, ndi mapuloteni angapo oyenda ngati MRP2. Kachiwiri, pali mitundu ya mitundu - Kusiyana kwapadera momwe nkhuku zimathandizira. Mwachitsanzo, rifampicin ndi indecracy yothandiza pa hepatocyte ya anthu ndi kalulu, koma alibe mphamvu pa rat hepacytes. Pomaliza, kuchepa kwapadera kwa hepatocyte yofiyira kumatha kuchepetsedwa kuchepetsedwa ma p450s ndikuwonjezereka. Monga taonera Chithunzi 3, hepatocytes kutsika pompopomphukira kuwonetsa gawo lotsika la cyp1a2, cyp2b6, ndi mayankho a Cyp3a4, koma mayankho amphamvu. Chifukwa chake, ma hepatocytes athanzi ku kachulukidwe woyenera woyenera ndi wofunikira kuti athe kudziwa zambiri.

Chithunzi 2. Chiyanjano pakati pa kupaka kachulukidwe ka mafupa obiriwira ndi enzyme

Source: Matekisikidwe Opeza Maphunziro aposachedwa, 2010, 7: 188 - 198

2). Kuyeserera kwa Hepatotot: Kuwona kusintha kwa Morphological pansi pa microgogs, monga cell morphology, phweule ndi lipid dontho kapena lipid loponi kapangidwe kake, ndi cell. Kuzindikira hepatocyte necrosis (monga momwe amasonyezera ndi Aminotransferase, archar dehydrogenase, ndi alanune aminotransferase) ndi apoptosis). Kwa cytokine - cytotoxicity, zikhalidwe zosiyanasiyana za hepatocytes sizinganenerepo ma cell omwe amasulidwa ku ceres, ma cell a Kipfylaidal endothelial.

3). "Njira yolumikizira" yophunzirira pang'onopang'ono yopanga mafuta ndi metabolites: Ntchito ya mankhwala - Michere yopukusira mu hepatocytes imayamba kuchepa pambuyo pa maola 24 akulemba. Pambuyo pakulowetsa hepatocyte zokhala ndi seramu - Free Medium wokhala ndi chidwi cha maola 24, sing'anga imasonkhanitsidwa ndikusakanikirana, kenako ndikusamutsidwa ku New Hepatocytes Yatsopano yophunzira (Chithunzi 3).

Chithunzi 3.

Source: Makalata a Centernabolism, 2016, 10: 3 - 15

3). Wunikirani ma cellular, endocytosis, endosomal kuthawa, komanso zolimbitsa thupi pa mtundu wa chiwongolero cha hepatocyte - adayika mankhwala a acid a nuclectic acid.

Chithunzithunzi Chotsatsira cha Sangwe

Mu vitro, hepatocytes amatha kukhala odziwika pakati pa zigawo ziwiri za collagen kapena Matrigel kuti ayanjanenso ndi magulu a Vivo, omwe amatchedwa sangweji. Hepatocytes ophatikizidwa pakati pa zigawo ziwiri za gel - collagen (sangweji (sangweji) imatha kusintha ma morphology ndi kusinthika kwa maselo, ndikusunga magwiridwe antchito kwa nthawi yayitali. Kuphatikiza apo, hepatocytes mu Chikhalidwe cha Sangweni amatha kubwezeretsa polarity, kulola kukhazikika koyenera kovomerezeka ndi zojambulajambula, komanso mapangidwe a network a Bile (Chithunzi 4).

Chithunzi 4. Mawonekedwe onyamula onyamula munthawi ya sakatocyte santelich

Source: Matekisikidwe Opeza Maphunziro aposachedwa, 2010, 7, 188 - 198

Mapulogalamu:

1. 1). Kuyerekezera kubisala kwa bilia.
2. 2). Kuwongolera kukwera kwa chiwindi ndi biliary kwa endo nati ndi ma metabolites ndi metabolites.
3. 3). Kuyerekeza kukonzekera kwa Cleading ndi kagayidwe ndi zonyamula, ndikupanga phyniology - zotengera pharmacokinetic mitundu.
4. 4). Kuphunzira hepatotoxicity ndi kupereka njira zamankhwala zothandizira matenda a chiwindi. Zambiri zimaphatikizidwa mu machitidwe a mankhwala amafanollogy kulosera za mankhwala - Kuvulala kwa chiwindi mwa anthu.
5.  
   1. 3. Hepatocyte 3D

6. Mu chikhalidwe cha 3D, hepatocyte amayamikiridwa mu atatu - mawonekedwe a matrix, omwe amafanana bwino ndi zochitika za Vivo a chilengedwe poyerekeza ndi zikhalidwe za Monolaal. Makina awa amalimbikitsa maselo - maselo ndi cell - Matrix Kugwirizana kwa chiwindi ntchito, kuphatikiza metabolism, protekin kasupe, ndi mapangidwe a bile. Zikhalidwe zoyambirira za Hepatocyte 3D zitha kugwiritsidwa ntchito pophunzira chiwindi - Ntchito zodziwika bwino kwambiri mu vivo - zokhala ngati zoyeserera zosokoneza bongo, kuwunika kwa poona, komanso kutengera matenda.

   Mtundu wa Sproid

Kudzera m'mabuku monga Ultra - Chikhalidwe chomata kwambiri, chizolowezi chomata zikhalidwe, ndi zikhalidwe 5), hepatocytes oyambira mpaka 150 - 175 popanda kudalira matrict a matriction. Ubwino umodzi wa chikhalidwe cha Spriod ndikuti spheroid iliyonse imangofuna 1,330 - maselo 2,000, kuchepetsa kuchuluka kwa maselo poyerekeza ndi njira zina zachikhalidwe cha 3D. Kafukufuku waposachedwa awonetsa kuti zikhalidwe zapamwamba za hepatocyte zimatha kukhala ndi milungu isanu, ndipo ma enzyme enzyme ntchito yopanda mankhwala, ma enzymes, komanso chizolowezi chimasungidwa bwino kwa masiku 14. Komabe, chiwindi ndi chovuta kwambiri kuposa kuphatikizika kwa cell. Mbali yakunyanja ya hepatocytes imalumikizana ndi magazi, pomwe bile imatuluka kuchokera kumbali ya apical, yomwe ili gawo lofunikira la kapangidwe ka chiwindi, ndipo izi sizingakhale zowonekeranso mu mtundu wa Sprohed.

Mapulogalamu:

1. 1). Kuwerenga pang'onopang'ono kupatsidwa chilolezo ndi ma metabolites.
2. 2). Kafukufuku wa hepatotoxicity.
3. 3). Kuyesa kwa ma cell okwera, endosososis, endosomal kuthawa, komanso zovuta zongocheza zomwe zakhazikitsidwa za hepatocyte - zida zamitundu yaying'ono ya nuclecyte.

Chithunzi 5. Njira yachikhalidwe cha Spriod

Chiwindi chamoyo

Kutanthauzira kogwirizana kwa zinthu ndi: Magawo 3d ochokera ku maselo a tsinde, ma cell othandizira, kapena maselo osiyanitse, omwe amatha kubereka minyewa yaing'ono mu vivo, cell - ku cell. Chiwindi cha chiwindi chadziwika kuti ndi chitsanzo chapamwamba kwambiri cha kafukufuku wa anthu amoyo.

Maselo a Maseri a Chiwindi Ntchito Yomanga:

Maselo a stralts a plum (ma pscs):

Ma cell a mtunda (asss) ndi kuyika maselo ang'onoang'ono a Sterhunt (IPSCS) kukhala ndi matenda ochulukirapo, pulasitiki, ndi mphamvu zopanda malire. Mothandizidwa ndi zifukwa zachindunji, amasiyanitsa hepatocyte - ngati maselo okhala ndi ntchito ndi ntchito. Komabe, zinthu za chiwindi zimachokera ku PSCs zimatha kusinthika kwa Epigenic ndi ma genetic, kuwonetsa chromosomal aneupidy amasintha.

Mapulogalamu:

1. 1). Ma genetic chiwindi
2. 2). Matenda Odwala Chiwindi
3. 3). Kuyesa kwa Cytotoxicity mankhwala

Minyewa ya chiwindi - Maselo Ochokera: Izi zimaphatikizapo anguocyte ndi hepatocyte. Kukhwima hepatocytes stem zomwe zingatheke komanso kuthekera kwa madera ena. Poyerekeza ndi PSC - zopangidwa ndi zinthu, zinthu zochokera ku minofu ya pulaimale zikuluzikulu zimakhala zokhazikika, komanso kukhala ndi ma genotypic komanso kukhazikika kwa mibadwo. Komabe, kutalika kwenikweni - mawu ophatikizira mphamvu ya hepatocyte zokhwima ndizochepa poyerekeza ndi fetal Hapacytes a Hepatocytes woyamba hepatocytes. Kukhala wamkulu mwachilengedwe Hepatocyte zachilengedwe kumakhala kovuta.

Njira Yachikhalidwe (Chithunzi 6): minofu ya chiwindi imagayidwa m'maselo amodzi, ndipo maselo osakanikirana a matrigel ndi ma cell aikidwa Mbale mu 24 - Sinthani mu cell kukhala chizinga chofukula (37 ° C) kwa mphindi 15. Pambuyo polimba, onjezani chikhalidwe chapadera. Kudutsa pafupifupi masiku 14. Sinthani sing'anga yoyambirira ndi sing'anga yosiyana ndi 7 - masiku 10.

Mapulogalamu:

1. 1). Mitundu ya hepatotoxicity
2. 2). Mu vita metabolism kusokonezeka kwa maphunziro
3. 3). Ndontho osamwa
4. 4). Kukula kwa mankhwala kwa benig komanso matenda a chiwindi

Chithunzi 6. Chikhalidwe ndi magawo a minofu - zopangidwa ndi chiwindi

Gwero: Cell & Bioscience (2023) 13: 197

Kuyerekeza ndi Chikhalidwe cha Spriod komanso chikhalidwe

4. Hepatocyte CO - Chikhalidwe Chachikhalidwe

1. 1. 2D yoyamba hepatocyte c - Chikhalidwe Chachikhalidwe
      

      Mu 2D yoyamba hepatocyte C - Model Model, mitundu iwiri kapena kuposerapo imasakanizidwa ndikuphatikizidwa mu ziwiri - Zikhalidwe. Gawo lofunikira la mtunduwu ndi kulumikizana mwachindunji pakati pa mitundu yosiyanasiyana ya maselo ndi matraclecrer matrix, kapena kutumiza kwa ma cytokines ndi mankhwala. Ntchito zoyambirira hepatocyte, monga kupanga mankhwala a Albumin ndi luso la kagayitso ka kagayidwe kazinthu, zimatha kusungidwa kwa milungu itatu.

      Mapulogalamu:

      1. 1). Hepatocytes wamkulu CO - Odziwika ndi Fibrobests: Mtunduwu umagwiritsidwa ntchito pophunzira chiletso cha chilolezo chochepa - mankhwala osokoneza bongo ndi metabolites.
      2. 2). Phiri yoyamba ya hepatocytes CO - Kuledzedwa ndi Zovala za parerymal chiwindi (E.G.)
      3. 3). Pulogalamu yoyamba ya hepatocytes CO - Maselo a T: Mtundu uwu umagwiritsidwa ntchito kudziwa chiwindi mankhwala - Maselo a TVU.
      4. 4). Hepatomax ™CO - Chikhalidwe Chaukhalidwe: Iphase zapanga CO - Chikhalidwe Chachikhalidwe chokhala ndi Hepatocytes Choyambirira Kuchokera Kumitundu Yosiyanasiyana, Amadziwika Kuti Ipatomax™. Pofika CO - Kukula kwa hepatocytes koyambirira kwa anthu okhala ndi maselo ozungulira, ndizotheka kugwiritsa ntchito mankhwala osokoneza bongo - ma enzyme ntchito mu hepatocytes ya anthu mpaka masabata atatu. Dongosolo ili ndi loyenera kuphunzira pang'onopang'ono - Misonkhano yolimbana ndi ma metabount ndi metabolites.

      Mtundu wa CO - Chikhalidwe cha chikhalidwe cha chikhalidwecho chimapereka chipilala choyenera cha mankhwala osokoneza bongo, poizoni, ndi chiwindi - Matenda ofananira - Matenda Ofananira
      
      

      3D yoyamba hepatocyte c - Chikhalidwe Chachikhalidwe

      Direct 3D CO - Chikhalidwe: Mtundu uwu umaphatikizapo kusakaniza mitundu iwiri kapena kupitilira apo ya chiwindi (E.g., ma cell a Spefcy, ophatikizidwa ndi zida zopangidwa ndi zida monga ma scagen, afayilo. Direct 3D CO - Chikhalidwe chimalola kuyanjana pakati pa ziwindi zosiyanasiyana kudzera pa cell - to rasion osinthira ma cytokines, kukhudzidwa motsatsa pakati pa hepatocyte.

      Mapulogalamu:

      1. 1). Mtundu wa chiwindi: ankakonda kuphunzira njirazo ndikupita patsogolo kwa chiwindi fibrosis.
      2. 2). Mankhwala - Kuvulala kwa chiwindi (Dili): kumathandiza kuti azitha kuwonongeka ndi matenda a chiwindi.
      3. 3). Kugwirizana ndi mankhwala osokoneza bongo, kagayidwe ka mankhwala, ndi ma enzyme ": kumayesa mankhwala osokoneza bongo, momwe amapangidwira, komanso momwe amapangidwira michere.

      Osavomerezeka 3D CO - Chikhalidwe: Njira iyi imagwiritsa ntchito njira yolekanitsira thupi (E.g. Kuyankhulana pakati pa maselo kumachitika kudzera pa ma cytokines.

      Mapulogalamu:
      Ogwiritsidwa ntchito pophunzira osayankhulana pakati pa maselo a chiwindi m'thupi.

      
      Mwachidule, monga mtsogoleri wa Vitro biologicalogicalogical, amapereka mayankho athunthu osakayeza mankhwala osokoneza bongo. Kuchokera ku chiwongola dzanja ndi chikhalidwe cha hepatocytes pulaneti yosiyanasiyana pakupanga zinthu zothandizirana ndi zinthu zina zofufuzira zofufuzira za vitro. Ndife anzathu omwe amadaliridwa kwa makasitomala m'makampani opanga mankhwala, odzipereka kuti ndife odula - Njira Zamapeto kwa Non - Kafukufuku Wakale.