{"id":45161,"date":"2023-01-17T02:51:56","date_gmt":"2023-01-17T00:51:56","guid":{"rendered":"https:\/\/umfcluj.ro\/cercetare\/centrul-de-cercetare-genomica\/platforma\/platforma-integrata-studii-in-vitro-organoizi-si-imagistica-moleculara\/"},"modified":"2024-04-01T01:56:09","modified_gmt":"2024-03-31T22:56:09","slug":"plate-forme-integree-pour-les-etudes-in-vitro-organoides-et-imagerie-moleculaire","status":"publish","type":"crct","link":"https:\/\/umfcluj.ro\/fr\/recherche\/centre-recherche-genomique\/plateforme\/plate-forme-integree-etudes-vitro-organoides-imagerie-moleculaire\/","title":{"rendered":"PLATE-FORME INT\u00c9GR\u00c9E POUR LES \u00c9TUDES IN VITRO &#8211; ORGANO\u00cfDES ET IMAGERIE MOL\u00c9CULAIRE"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-post\" data-elementor-id=\"45161\" class=\"elementor elementor-45161 elementor-13023\" data-elementor-post-type=\"crct\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-4f33edf0 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"4f33edf0\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-1eab6f08\" data-id=\"1eab6f08\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-48c5195a elementor-widget elementor-widget-text-editor\" data-id=\"48c5195a\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t\t\t\t\t\t<p><strong>Modele <em>in vitro<\/em><\/strong>. Studiile pot fi efectuate pe linii celululare aderente, c\u00e2t \u0219i suspensie, at\u00e2t \u00een condi\u021bii normale sau hipoxice. De asemenea, se pot dezvolta \u015fi evalua prin diferite studii, culturi celulare 3D precum sferoizi, organoizi, co-culturi, etc.<\/p><p>Testele de evaluare a prolifer\u0103rii celulare, a viabilit\u0103\u021bii celulare \u0219i a citotoxicit\u0103\u021bii sunt utilizate \u00een mod curent pentru a monitoriza efectul diferitelor tratamente.<a href=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-7.jpg\"><img fetchpriority=\"high\" decoding=\"async\" class=\"alignright wp-image-13387 size-medium\" src=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-7-300x259.jpg\" alt=\"\" width=\"300\" height=\"259\" srcset=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-7-300x259.jpg 300w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-7-1024x885.jpg 1024w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-7-768x664.jpg 768w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-7.jpg 1437w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a><br \/>Pentru r\u0103spunsul la diferite scenarii de tratament \u0219i \u00een cazul \u00een care se utilizeaz\u0103 doze multiple, putem calcula parametrul toxicologic standard IC50 prin:<\/p><ul><li><strong>MTT<\/strong> un test colorimetric pentru evaluarea activit\u0103\u021bii metabolice a celulelor (utilizat pentru celule aderente);<\/li><li><strong>MTS<\/strong> este utilizat pentru a evalua proliferarea celular\u0103, viabilitatea celular\u0103 \u0219i citotoxicitatea &#8211; specific\u0103 pentru celulele \u00een suspensie;<\/li><li>este foarte stabil \u0219i prezint\u0103 citotoxicitate mic\u0103. Timpul de incubare este mai lung (24 p\u00e2n\u0103 la 48 de ore) &#8211; at\u00e2t pentru celulele aderente, c\u00e2t \u0219i pentru cele \u00een suspensie.<\/li><\/ul><p>Modificarile proceselor celulare \u0219i moleculare \u00een celulele studiate pot fi evaluate cu ajutorul urm\u0103toarelor teste func\u021bionale:<\/p><p><a href=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-9_.jpg\"><img decoding=\"async\" class=\"alignleft wp-image-13398 size-medium\" style=\"margin-right: 2em;\" src=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-9_-300x292.jpg\" alt=\"\" width=\"300\" height=\"292\" srcset=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-9_-300x292.jpg 300w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-9_-1024x995.jpg 1024w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-9_-768x747.jpg 768w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-9_.jpg 1469w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a><\/p><ul><li><strong>Evaluarea apoptozei<\/strong> folosind dubl\u0103 colorare (TMRE\/Hoechst pentru nucleu \u0219i poten\u021bial mitocondrial, respectiv Annexin-V\/PI pentru nucleu \u0219i membrana celular\u0103) prin microscopie cu fluorescen\u021b\u0103 \u015fi cu ajutorul cititorului automat de pl\u0103ci CELIGO.<\/li><li><strong>Evaluarea autofagiei<\/strong> folosind monodansil cadaverina \u0219i iodura de propidiu pentru colorarea vacuolele autofagice, respectiv nucleul \u0219i evaluarea prin microscopie cu fluorescen\u021b\u0103 \u015fi cu ajutorul cititorului automat de pl\u0103ci CELIGO.<\/li><li><strong>Testul de colonie<\/strong> (testul clonogenic) este considerat a fi standardul de aur pentru evaluarea r\u0103spunsului in vitro al celulelor la un compus terapeutic.<\/li><li><strong>Testul scratch<\/strong> (testul de vindecare a r\u0103nilor) este o tehnic\u0103 standard in vitro pentru analiza migr\u0103rii celulare. Se poate evalua prin microscopie confocal\u0103 \u015fi cu ajutorul cititorului automat de pl\u0103ci CELIGO.<\/li><\/ul><p>\u00a0<\/p><p><strong>Citometrie \u00een flux<\/strong> permite caracterizarea popula\u021biilor de celule \u0219i m\u0103soar\u0103 simultan propriet\u0103\u021bile optice inerente ale celulelor \u0219i ale anumitor fluorofori lega\u021bi de anticorpi care s-au legat de celule. Cu ajutorul acestora, putem evalua:<\/p><p><a href=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-12.jpg\"><img decoding=\"async\" class=\"alignright wp-image-13391 size-thumbnail\" src=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-12-150x150.jpg\" alt=\"\" width=\"150\" height=\"150\" \/><\/a> <a href=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-11.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignright wp-image-13392 size-thumbnail\" src=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-11-150x150.jpg\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/p><ul><li><strong>Apoptoza<\/strong> cu ajutorul colora\u021biei Annexin-V\/PI pentru a identifica stadiile timpurii, t\u00e2rzii \u0219i necrotice ale procesului de moarte celular\u0103.<\/li><li><strong>Ciclul celular<\/strong> cu ajutorul colora\u021biei cu iodur\u0103 de propidiu pentru a analiza fazele subG0, G1, S, G2\/M.<\/li><li><strong>Caracterizarea celulelor stem<\/strong> prin intermediul anticorpilor CD11b, CD29, CD34, CD44, CD45, CD73.<\/li><\/ul><p><strong>Exozomii<\/strong>, vezicule cu o dimensiune cuprins\u0103 \u00eentre 30-150 nm, pot fi izola\u021bi din diferite fluide biologice (medii de cultur\u0103 celular\u0103, plasm\u0103, saliv\u0103, ser etc.). Aceste vezicule sunt entit\u0103\u021bi remarcabile care \u00eenglobeaz\u0103 molecule bioactive importante \u0219i pot fi utilizate ca poten\u021biali biomarkeri pentru diagnosticare, prognostic \u0219i r\u0103spuns la terapie.<a href=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-13.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignright wp-image-13393 size-medium\" src=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-13-300x228.jpg\" alt=\"\" width=\"300\" height=\"228\" srcset=\"https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-13-300x228.jpg 300w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-13-1024x777.jpg 1024w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-13-768x583.jpg 768w, https:\/\/cdn.umfcluj.ro\/uploads\/2023\/01\/image-13.jpg 1221w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a><\/p><ul><li>Izola\u021bi prin ultracentrifugare<\/li><li>Caracterizare fizic\u0103 \u0219i molecular\u0103: NanoSight NS300, Western Blot<\/li><li>qRT-PCR, NGS<\/li><\/ul><p><strong>Testarea noilor compu\u0219i terapeutici<\/strong> este utilizat\u0103 intensiv \u00een laboratorul de culturi celulare. Compu\u0219ii sunt selecta\u021bi \u00een func\u021bie de o semn\u0103tur\u0103 muta\u021bional\u0103 specific\u0103 \u0219i prin efectuarea mai multor teste func\u021bionale (viabilitate celular\u0103, modific\u0103ri morfologice, migra\u021bie, invazie) pentru a evalua procesele celulare \u0219i moleculare importante.<\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-80f9f7f elementor-widget elementor-widget-gallery\" data-id=\"80f9f7f\" data-element_type=\"widget\" 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INT\u00c9GR\u00c9E POUR LES \u00c9TUDES IN VITRO - ORGANO\u00cfDES ET IMAGERIE MOL\u00c9CULAIRE - UMF Cluj<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/umfcluj.ro\/fr\/recherche\/centre-recherche-genomique\/plateforme\/plate-forme-integree-etudes-vitro-organoides-imagerie-moleculaire\/\" \/>\n<meta property=\"og:locale\" content=\"fr_FR\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"PLATE-FORME INT\u00c9GR\u00c9E POUR LES \u00c9TUDES IN VITRO - ORGANO\u00cfDES ET IMAGERIE MOL\u00c9CULAIRE - UMF Cluj\" \/>\n<meta property=\"og:description\" content=\"Modele in vitro. 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