Na farko hepatocytes: kayan aiki mai mahimmanci don ci gaba ba tare da bincike ba a Binciken Magunguna

Na farko hepatocytes: kayan aiki mai mahimmanci don ci gaba ba tare da bincike ba a Binciken Magunguna

Matsakaicin firam na farko don bayyanar miyagun ƙwayoyi, yana taka muhimmiyar rawa a cikin metabolism na miyagun ƙwayoyi da guba. Primal Hepatocytes suna da cikakken bakan halayyar salula da matakan enzymes da kuma cofacolory, suna daure enabymus) da enzymes na cytostmes kamar hadewar metaboles) da aka samu enaboles da aka samo a hanta. A saboda wannan dalili, ana daukar na farko hepatocytes a matsayin ma'aunin zinare don ginin hanta na Vitro kuma ana yaba shi da masu binciken miyagun ƙwayoyi, da kuma nazarin miyagun ƙwayoyi, da kuma nazarin miyagun ƙwayoyi, da kuma nazarin miyagun ƙwayoyi. Wannan labarin yana ba da taƙaitaccen tsarin 2D da 3D bisa ga tsarin heckatocytes da aikace-aikacen su a ci gaban magunguna.

Keywords: Keywords: Keywords: Prim Hepatocytes, 2D Namo, Namo, Kono, CO - Al'adu.

1. 1. Ware na farko hepatocytes

Kafuwar heematocytses na mahimmancin mataki ne a cikin tsarin hanta na vitro, tare da biyu - suttukan collgenase passion hanyar kasancewa mafi yawan amfani. A cikin ƙananan dabbobi, ana iya yin tursasawa hanta ta hanyar tashar jirgin ruwa ko na ƙasa da ke fi da girma yawanci suna buƙatar tursasawa ta hanyar hanta ko sassan. Abubuwa da yawa sun shafi wadatar da hepatocytes na hepatocytes: na farko, Collogase yakamata ya kasance ba - cytotoxic. Na biyu, lokacin narkewa yana da mahimmanci-Dukansu suna ƙarƙashin - narkewa da sama da - narkewa na iya magance yawan amfanin gona da ƙarfi. Na uku, yanayin hanta dole ne ya zama mafi kyau duka, kamar yadda hepatocytes suna da matukar kula da lalacewar ischemic. An yi amfani da hanta don shiri na hepatocyte cikin sauri don rage farashin rayuwa da hana hypocia hypexiya da kuma m ishemia m.

Ka'idojin na farko na hepatocytes da aka yi amfani da su a binciken magunguna sune kamar haka:

1. 2. Na farko hepatocyte al'adu

Dakatarwar hepatocyte

Ya ƙunshi ƙwayoyin cuta - tsirar da enzymes da masu aikin enzymes da cockactors, sanya shi dace da nazarin jagorancin hanyoyin rayuwa daban-daban. Koyaya, mafi yawan dakatar da dakatarwar hepatocytes da ayyukan miyagun ƙwayoyi - mencymes a hankali raguwa kamar yadda a cikin pitro shiryayye zuwa mafi yawan 4 hours. Wannan ƙirar ana amfani da ita don kimanta tabbacin magunguna tare da matsakaici zuwa farashin tsawan kuɗi. Lokacin da aka ƙaddara ta share ƙasa ta ƙasa da kashi 20%, ingantaccen ƙimar ba za a iya tantance ƙimar ba. Darewar Hepatocyte - An kafa shi ne a cikin ƙirar metabolism na vitro ba su da isasshen halayen rayuwa na rayuwa don yin hasashen tsarin tsabtace da kayayyakin metabolism na waɗannan mahaɗan. Dakatar da hanyar hepatocyte ta hanyar (Hoto 1) don mika lokacin shiryawa zuwa awanni 20 ko kuma ya fi tsayi.

Aikace-aikacen:Ayyukan Enzyme da kuma bincike mai gamsarwa na rayuwa don ƙwayoyin kwayoyi.

Hoto 1

Source: Manyan Metab Investab, 2012,40 (9): 1860-1865

Model mai ƙarfi Hepatocytes

Priminal hepatocytes ana da hade a cikin tsarin 2D a kan Collagen - mai cike fararen al'adu. Hepatocytes suna nuna yanayin miyayya, tare da nuclei na haifar, galibi suna gabatar da shi a cikin lancleated tsari. Runduna na al'adun hepatocyte monolaocyte sun hada da: 1. Canjin polarity da aiki.2. Rashin wasu nau'ikan kwayar halitta (I.e., ba - sel na parchymy ba) waɗanda ake buƙata don aikin al'ada.3. Rashin iya samar da isasshen abubuwan da suka dace da abubuwan fastoci don tallafawa hepatocytes a cikin ayyukansu (kamar bile acid da kuma samar da furotin biosynthesis).

Aikace-aikace:

1). Kimanin magani - hulda da miyagun ƙwayoyi:Wannan ya hada da shigowar enzyme, inzhiition enzyme, da kuma binciken sufuri. Da fari dai, lokacin da magani yake yi a matsayin wanda Induecer, zai iya haifar da haɓaka faɗakarwa na ƙwayoyi - menzymes enzymes da sufuri. Mai ƙarfi da bashi zai iya samar da kwayoyin halittu da yawa lokaci guda, kamar sahunnbiyar sahunnan -3, Cyp3C9, UGT, da kuma sunadarai masu jigilar sufuri kamar MrP2. Abu na biyu, akwai nau'ikan bambance-bambance a cikin yadda hepatocytes amsa da shigarwar. Misali, Grampicicicicicicicicicicic ne mai tasiri wanda aka kirkira don ɗan adam da zomo heepatocytes, amma bashi da sakamako sakamakon a bera hepatocytes. A ƙarshe, ƙaddamar da ɗumbin shararar da hepatocytes na iya haifar da rage girman girman P450s da kuma karuwa da martani na baya. Kamar yadda aka nuna a cikin Hoto na 3, hepatocytes a kasa plating na kasa suna nuna ƙananan ayyukan muhimmi na cyp1a2, cyp3a4, amma abin da ya fi karfin sakonni. Saboda haka, lafiya hepatocyteses a wani yanki mai da ya dace ya zama dole don samun bayanan da suka dace na ilimin ta da suka dace.

Adadi na 2

Source: Fasahar Gano ta yanzu, 2010, 7: 188 - 198

2). Gwajin Hepatotoxi: Lura da canje-canjen ilimin cututtukan cututtukan cututtukan cututtukan cututtukan cututtukan cututtuka, kamar su ilimin halittar jiki, ciyawar da lipid dropolet, da kuma abin da aka makala / cire. Gano na hepatocyte necrosis (kamar yadda aka nuna ta hanyar aspartate Aminotransnantasen, Lectate dhydrogenase, da alanin goinotransansronnase) da apoptosise (DNine Formentise). Don Cytokine - A tsarin kula da Hepatocytes, al'adun monatocytes na hepatocytes ba zai iya hasashen martani ba saboda sel na Kuffyms, kamar sel na Kufffer, da sel na Sinusial, da sel na Sinusial, da sel na Sinusial.

3). "Hanyar Relay" don nazarin sannu da hankali: Ayyukan Magunguna - enzymes enzymes a cikin shepatocytes mai sanyin gwiwa yana fara raguwa bayan sa'o'i 24 na plating. Bayan in kula da hepatocytes da Magana - Matsakaicin matsakaici wanda ke ɗauke da mahimman bayanai masu ban sha'awa na sa'o'i 24, ana tattara matsakaici zuwa sabon hepatocytes don ƙarin nazari (Hoto 3).

Hoto na 3. Mai siyar da Hepatocyte Relatol

Source: Harafin mitabolism na miyagu, 2016, 10: 3 - 15

3). Kimanta Uptakewar salula, entocytosis, tserewa mai ƙarewa, da kuma shafe tasirin gene na hepatocyte - An yi niyya kananan kwayoyi masu nobini.

Tsarin Noma

A cikin Vitro, hepatococytes za a iya rarrabawa tsakanin yadudduka biyu na Collagen ko Matrigel don sake gina a cikin tsarin vivo, wanda aka sani da Sandwich Noma. Hepatocytes da hade tsakanin yadudduka biyu na gel - Collagen (sanwic tsarin) na iya inganta ilimin ilimin halittar da mafi kyawun sel, kuma kula da aikinsu na tsawon lokaci. Bugu da ƙari, hepatocytes a cikin sanwic al'ada na iya mayar da polarity, yana barin ingantacciyar rarrabuwa ta masarauta da canalisular dattin dects (Hoto na 4).

Hoto 4

Source: Fasahar Gano ta yanzu, 2010, 7, 188 - 198

Aikace-aikace:

1. 1). Kimanta bincike na BilIES na mahadi.
2. 2). Kimanta tsarin hefatic da beliiye na ƙwararrun ƙwayoyin cuta da abubuwan da suka faru da metabolites.
3. 3). Ana daidaita ma'anar ma'anar Metabolism da masu sufuri da sufuri, da kuma gina ilimin kimiyyar lissafi - tushen tsarin cinikin.
4. 4). Nazarin hefatotoxicity da samar da hanyoyin motsa jiki ga cututtukan asibiti - Raunin hanawa ya jawo shi. An haɗa bayanan da aka haɗa cikin samfuran magunguna na tsarin da za a iya hango hasƙiyar ƙwayoyin cuta - Haƙiƙa ya ji rauni a cikin mutane.
5.  
   1. 3. Abu na farko Hepatocyte 3d Al'adu

6. A cikin tsarin al'adu na 3D, hepatocytes a cikin uku - matrix matrix guda uku, wanda mafi kyau a cikin tsarin hanta na vivo an kwatanta shi da al'adun Monolola. These systems promote cell-cell and cell-matrix interactions, which can restore more of the liver’s physiological functions, including drug metabolism, protein secretion, and bile formation. Za'a iya amfani da al'adun heckatocyte 3d don yin nazarin hanta - takamaiman ayyuka a cikin ƙari a vivo - suna bayar da fa'idodi don gwajin miyagun ƙwayoyi, ƙididdigar rashin cancanta, da kuma ƙimar guba.

   Sperid samfurin

Ta hanyar dabaru kamar + - al'adun da aka makala da aka makala, da al'adun tantanin tarihi (Hoto na 5), ​​hepatocytes na farko da diamitaid na har zuwa 150 -, μm ba tare da dogaro da matrices na waje ba. Daya fa'idodin al'adu na sphoid shine kowane mai fafutuka ne kawai 1,330 - 2,000 sel, yana rage yawan kwayoyin idan aka kwatanta da sauran dabarun al'adun 3. Nazarin kwanan nan sun nuna cewa al'adun farko na hepatocyte na farko zasu iya ci gaba da har zuwa lokacin da Cyp Enzyme ya da musayar su tsawon kwanaki 14 a cikin al'adun sphoidoid. Koyaya, hanta ya fi rikitarwa fiye da tara. Theanƙwasa gefen hepatocytes yayi ma'amala da jini, yayin da bile yake gudana daga apical gefen, wanda shine mabuɗin mahimmin sandar hanta lullu, kuma wannan ba tukuna iya zama a cikin samfurin speridid.

Aikace-aikace:

1. 1). Nazarin jinkirin yin tarihin tsallakewar fili da metabolites ɗinsa.
2. 2). Binciken Hepatotoxity.
3. 3). Kimiyya ta UPTake, engocytosis, tserewa mai tsewa, da kuma sharar tasirin tasirin kwayoyin hepatocyte - Manyan kwayoyi masu nobini.

Hoto na 5. Hanyar al'adu

Haɗin hanta na ƙira

Ma'anar yarjejeniya na Orpens shine: Tsarin 3D da aka samo daga sel, sel na gari, ko kuma bambance bambancen ƙwayoyin cuta a cikin vivo microenvirement da sel mai mahimmanci. An gano gabobin hanta a matsayin samfurin ci gaba don Binciken Halin Halin Halin Halin Hearch.

Tushen tantanin halitta don hanta na hanta:

① Pleuripotent Stell Sell (PSCs):

Kwayoyin sel na kara (ESCS) da jawo Plouripotent Sells (ipscs) sun mallaki pluripotenc mai girma, filastik, da ƙarfin yuwuwar da ba a iyakance ba. A karkashin tasirin takamaiman abubuwan da suka dace, sun bambance cikin hepatocyte - kamar sel tare da aiki da aiki. Koyaya, gabobin hanta sun samo asali ne daga PSCs na iya yin canje-canje da gyaran kwayoyin cuta, suna nuna alamun yanayin ƙwaƙwalwar ƙwaƙwalwar ƙwaƙwalwar ƙwaƙwalwar ƙwaƙwalwar ƙwaƙwalwar ƙwaƙwalwa ta canza yayin amsar.

Aikace-aikace:

1. 1). Ganowar hanta na hanta
2. 2). Motocin hanta na kamuwa da cuta
3. 3). Gwajin cytotoxiity

Matan karar - Sellar da aka samo: Waɗannan sun haɗa da cholangiocytes da hepatocytes. Balagagge hepatocytes yana riƙe da karfin tantanin halitta da iyawa sosai a cikin takamaiman mahalli. Idan aka kwatanta da PSC - Abubuwan da suka samo asali ne daga ƙwayar firamare sun yi girma, tare da ƙarin ƙwayoyin cuta, da kuma kula da abubuwan gani da kwayoyin halitta a cikin al'ada. Koyaya, tsawon lokacin da za a iya amfani da karfin rai na yau da kullun na ɗan adam. Kulla da tsofaffin hepatocyte na gabaɗaya ya kasance ƙalubale.

Hanyar al'adu (Hoto na 6): an narkar da nama na hanta a cikin sel guda, da cakuda matrigel da ƙwayoyin seeded a cikin 24 - tsarin da aka fasalta. Acubate a cikin sel al'adu (37 ° C) na mintina 15. Bayan haka, ƙara takamaiman matsakaici na al'ada. Nassi bayan kimanin kwanaki 14. Sauya ainihin matsakaici tare da matsakaici na bambance-bambance bayan 7 - kwanaki 10.

Aikace-aikace:

1. 1). Tsarin Hepatotoxitity
2. 2). A cikin rikice-rikice na Vitro Metabolism
3. 3). Ban da giya
4. 4). Ci gaban Magunguna don Benign da cutar sankarau

Hoto 6. Al'adu da nassi tsari na nama - Kungiyoyin hanta sun samo asali

Source: Weel & BIOSCENCEN (2023) 13: 197

Kwatanta al'adun gargajiya da al'adun gargajiya

4. Tsarin Hepatocyte Co - Model na al'adu

1. 1. 2d farko hepatocyte Co - samfurin al'adu
      

      A cikin firam na farko na hepatocyte Co - Model na al'adu, nau'ikan sel biyu suna gauraye da ladabi a cikin biyu - yanayin girma. Babban fasalin wannan ƙirar ita ce ma'amala ta kai tsaye tsakanin nau'ikan sel daban-daban, hulɗa tsakanin sel da kuma matrix da ke musun ta hanyar cytounes da hanyoyin sadarwa na akuya. Farkon heepatocyte ayyuka, kamar albashin Album da ikon metabolism, za a iya kiyaye har zuwa makonni uku.

      Aikace-aikace:

      1. 1). Priminal hepatocytes Co - An yi amfani da Fibroblasts: Ana amfani da wannan ƙirar don nazarin shakkun da sauri - metabzing mahadi da metabolites da metabolites da metabolites.
      2. 2). Priminal hepatocytes Co - A hadu da ba - Patharinta Sells Sells (ELG., Sel Sells, Kwayoyinta na Ingantaccen Lafiya, Sokin Ba da amfani ga Matsayi na Cytiyilar hanawa bayan bayyanar miyagun ƙwayoyi.
      3. 3). Primine hepatocytes Co - A haddacece Tells: An yi amfani da wannan ƙirar don gano ƙwayoyin cuta miyagun ƙwayoyin cuta - takamaiman martani.
      4. 4). Hepatomax ™CO - Tsarin al'ada: Iphase ya kirkiro da CO - tsarin al'ada tare da hepatocytes na farko, wanda aka sani da hepattix™. By Co - Culuringing Premilytesan Adam Hepatocytes tare da sel stomal, yana yiwuwa a kula da kyakkyawan magani - metabolia cikin aikin enzyme na tsawon makonni 3. Wannan tsarin ya dace da nazarin jinkirin - Metabolizing fili tsinkaye da metabolites.

      Wannan co - Model na al'adu yana ba da ƙarin dandamali na ilimin halin ciki don tantance miyagun ƙwayoyi, masu guba, da hanta - suna bayar da fahimta cikin yadda nau'ikan tantanin halitta daban-daban.
      
      

      3d farko hepatocyte Co - samfurin al'adu

      Kai tsaye 3d co - Al'adu: Wannan samfurin ya shafi haɗuwa da ƙwayoyin hanta biyu ko fiye, ƙwayoyin sel, sel mai sauƙi, sel steels, selpin, selglate sel collagen, fibrin, selminate sel collagen, fibrin, selminate, ko hydrogels. Direct 3D CO - Al'adu yana ba da damar kusanci tsakanin sel na hanta daban-daban ta hanyar ƙirar sel, siginar matrix na matrix, da kuma saurin tarihin matrix, da kuma inganta sadarwa tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes tsakanin hepatocytes.

      Aikace-aikace:

      1. 1). Fribrosis hanta: An yi amfani da nazarin hanyoyin da cigaban hanta fibrosis.
      2. 2). Magunguna - Sanarwa da hanta ta haifar da tsarin hanta: Taimaka hoto da kuma tantance lalacewar hanta da kwayoyi suka haifar.
      3. 3). Cikakken lissafin miyagun ƙwayoyi, da kuma shigar da enzyme: kimanta yadda magunguna daban-daban suke hulɗa, da kuma yadda suke hade da enzymes.

      Kai tsaye 3D CO - Al'adu: Wannan hanyar tana amfani da tsarin rabuwa ta zahiri (E.G., transwell ko wasu kayan) zuwa al'adun NIH / 3T3 ko Semusoidal Sells na NIH / 3T3, inda sel na uku, inda sel na uku, an hana lambar sirri tsaye. Sadarwa tsakanin sel na faruwa ta hanyar narkewa mai narkewa.

      Aikace-aikace:
      Amfani da nazarin ba - tuntuɓar sadarwar tsakanin sel hanta a cikin jiki.

      
      A taƙaice, Iphayan, a matsayin jagora a cikin binciken bincike na vitro, yana ba da cikakkun hanyoyin da ba shi da magani ba. Daga ware da al'adun farko hepatocytes daga ci gaban tallafi kayayyakin kamar takamaiman aikace-aikacen bincike don gano abubuwan bincike na comro na bincike don gano ƙwayoyi da ci gaba. Mu ne amintaccen abokin ciniki ne ga abokan ciniki a masana'antar harhada magunguna, sun dage wajen samar da yankan - gefen mafita don ba - bincike na asibiti ba.