{"id":1811,"date":"2026-01-09T12:39:26","date_gmt":"2026-01-09T11:39:26","guid":{"rendered":"https:\/\/bioeducator.eu\/?page_id=1811"},"modified":"2026-01-09T14:10:08","modified_gmt":"2026-01-09T13:10:08","slug":"metody-odklejania-komorek","status":"publish","type":"page","link":"https:\/\/bioeducator.eu\/?page_id=1811","title":{"rendered":"Metody odklejania kom\u00f3rek"},"content":{"rendered":"\n<div class=\"wp-block-columns is-layout-flex wp-container-core-columns-is-layout-8f761849 wp-block-columns-is-layout-flex\">\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<p class=\"wp-block-paragraph\"><a href=\"https:\/\/bioeducator.eu\/?page_id=1801\">Pasa\u017cowanie kom\u00f3rek<\/a> jest rutynowo wykonywan\u0105 czynno\u015bci\u0105. Kom\u00f3rki rosn\u0105ce w zawiesinie nie stwarzaj\u0105 problem\u00f3w.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Wyb\u00f3r odpowiedniej metody odklejania kom\u00f3rek od pod\u0142o\u017ca (tzw. dysocjacji) zale\u017cy od typu linii kom\u00f3rkowej oraz celu dalszych bada\u0144. Niekt\u00f3re bia\u0142ka powierzchniowe s\u0105 niezwykle wra\u017cliwe na dzia\u0142anie enzym\u00f3w, co wymusza stosowanie \u0142agodniejszych alternatyw.<\/p>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\">\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"683\" src=\"https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-Cdt3s_jp1OY-unsplash-1024x683.jpg\" alt=\"pasa\u017cowanie kom\u00f3rek\" class=\"wp-image-1804\" srcset=\"https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-Cdt3s_jp1OY-unsplash-1024x683.jpg 1024w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-Cdt3s_jp1OY-unsplash-300x200.jpg 300w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-Cdt3s_jp1OY-unsplash-768x512.jpg 768w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-Cdt3s_jp1OY-unsplash-391x260.jpg 391w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-Cdt3s_jp1OY-unsplash-1536x1025.jpg 1536w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-Cdt3s_jp1OY-unsplash-2048x1366.jpg 2048w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n<\/div>\n<\/div>\n\n\n\n<h2 class=\"wp-block-heading\">Metody enzymatyczne<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">To najpowszechniejsza grupa metod, wykorzystuj\u0105ca proteazy do trawienia bia\u0142ek adhezyjnych.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Trypsyna-EDTA:<\/strong> Trypsynizacja jest najcz\u0119\u015bciej stosowan\u0105 metod\u0105. Trypsyna trawi bia\u0142ka adhezyjne uwalniaj\u0105c kom\u00f3rki. EDTA wspiera jej dzia\u0142anie. Chelatuje ono jony Ca<sup>2+<\/sup> i Mg<sup>2+,<\/sup> kt\u00f3re s\u0105 kofaktorami dla bia\u0142ek adhezyjnych (jak kadheryny). Dzi\u0119ki temu zminimalizowane jest zbijanie si\u0119 kom\u00f3rek w agregaty.<\/li>\n\n\n\n<li><strong>Kolagenaza:<\/strong> Stosowana cz\u0119\u015bciej przy izolacji kom\u00f3rek bezpo\u015brednio z tkanek (np. z tkanki t\u0142uszczowej czy guza), poniewa\u017c skutecznie trawi macierz zewn\u0105trzkom\u00f3rkow\u0105 (ECM).<\/li>\n\n\n\n<li><strong>Dispaza:<\/strong> Jest to \u0142agodna proteaza bakteryjna. Cz\u0119sto u\u017cywana, gdy chcemy odklei\u0107 ca\u0142\u0105 warstw\u0119 kom\u00f3rek (tzw. <em>cell sheet<\/em>) bez rozbijania ich na pojedyncze jednostki.<\/li>\n<\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Metody nieenzymatyczne (chemiczne)<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Trypsyna trawi nie tylko bia\u0142ka adhezyjne ale r\u00f3wnie\u017c bia\u0142ka b\u0142onowe i receptory. Je\u015bli w badaniu niezb\u0119dne jest ich funkcjonowanie a czas hodowli uniemo\u017cliwia ich odbudow\u0119 (np. gdy odrywamy kom\u00f3rki w celu przeprowadzenia analizy cytometri\u0105 przep\u0142ywow\u0105), zastosowanie trypsyny jest niemo\u017cliwe. Mo\u017cna wtedy wykorzysta\u0107 techniki chemiczne lub mechaniczne.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Roztwory oparte na EDTA\/EGTA:<\/strong> Wykorzystuj\u0105 wy\u0142\u0105cznie mechanizm chelatowania jon\u00f3w miedzi i magnezy. Kom\u00f3rki trac\u0105 przyczepno\u015b\u0107 &#8222;poluzowuj\u0105c&#8221; swoje wi\u0105zania z pod\u0142o\u017cem, co jest znacznie mniej inwazyjne ni\u017c trawienie bia\u0142ek.<\/li>\n<\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Metody mechaniczne<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Czasami wykorzystuje si\u0119 techniki mechanicznego odrywania kom\u00f3rek od pod\u0142o\u017ca. W zale\u017cno\u015bci od rodzaju kom\u00f3rek dost\u0119pne s\u0105 dwie techniki:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Skrobanie (Scraping):<\/strong> U\u017cycie gumowych lub plastikowych skrobaczek (<em>cell scrapers<\/em>). Metoda ta jest szybka, ale cz\u0119sto powoduje liz\u0119 cz\u0119\u015bci kom\u00f3rek, co uwalnia DNA i mo\u017ce powodowa\u0107 &#8222;zlepianie si\u0119&#8221; zawiesiny. Mo\u017ce by\u0107 przydatna w badaniu bia\u0142ek b\u0142onowych. Skrobanie ma niestety wad\u0119. Jego stosowanie powoduje zwi\u0119kszenie odsetka martwych kom\u00f3rek.<\/li>\n\n\n\n<li><strong>Wyp\u0142ukiwanie (Flushing):<\/strong> W przypadku linii s\u0142abo adherentnych (np. HEK293), wystarczy energiczne skierowanie strumienia po\u017cywki na dno naczynia.<\/li>\n<\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Stosowanie specjalnych naczy\u0144 hodowlanych<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Istniej\u0105 specjalnie modyfikowane naczynia hodowlane (np. systemy Nunc\u2122 UpCell\u2122), kt\u00f3rych powierzchnia zmienia w\u0142a\u015bciwo\u015bci hydrofobowe w zale\u017cno\u015bci od temperatury.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>W <strong>37\u00b0C<\/strong> powierzchnia jest hydrofobowa i sprzyja adhezji.<\/li>\n\n\n\n<li>Po obni\u017ceniu temperatury do <strong>20\u201325\u00b0C<\/strong> powierzchnia staje si\u0119 hydrofilowa, co powoduje samoczynne odklejenie si\u0119 kom\u00f3rek bez u\u017cycia jakichkolwiek odczynnik\u00f3w.<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">To rozwi\u0105zanie sprawdzi si\u0119 idelanie do odrywania kom\u00f3rek bez naruszania struktury wytworzonej przez nie b\u0142ony.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Podsumowanie wyboru metody:<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Podsumowuj\u0105c, najcz\u0119\u015bciej stosowana trypsynizacja ma swoje wady i czasami trzeba j\u0105 zast\u0105pi\u0107 innymi metodami.<\/p>\n\n\n\n<figure class=\"wp-block-table\"><table class=\"has-fixed-layout\"><tbody><tr><td><strong>Metoda<\/strong><\/td><td><strong>Zalety<\/strong><\/td><td><strong>Wady<\/strong><\/td><\/tr><tr><td><strong>Trypsynizacja<\/strong><\/td><td>Bardzo wysoka wydajno\u015b\u0107, szybko\u015b\u0107.<\/td><td>Mo\u017ce niszczy\u0107 receptory powierzchniowe.<\/td><\/tr><tr><td><strong>Chelatowanie (EDTA)<\/strong><\/td><td>Zachowuje struktur\u0119 bia\u0142ek b\u0142onowych.<\/td><td>Wolniejsze, nieefektywne przy silnie adherentnych liniach.<\/td><\/tr><tr><td><strong>Skrobanie<\/strong><\/td><td>Brak odczynnik\u00f3w chemicznych.<\/td><td>Wysoki odsetek martwych<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n<h2 class=\"wp-block-heading\">\u0179r\u00f3d\u0142a<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Literatura:<\/p>\n\n\n\n<ol class=\"wp-block-list\">\n<li><a href=\"https:\/\/www.atcc.org\/resources\/culture-guides\/animal-cell-culture-guide#Cell\">ATCC<\/a><\/li>\n\n\n\n<li>Hodowla kom\u00f3rek i tkanek in vitro. P.red. Stanis\u0142awy Stok\u0142osowej; 2004<\/li>\n\n\n\n<li><a href=\"https:\/\/www.thermofisher.com\/\">https:\/\/www.thermofisher.com\/<\/a><\/li>\n<\/ol>\n\n\n\n<p class=\"wp-block-paragraph\">Obrazy:<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Photo by <a href=\"https:\/\/unsplash.com\/@niaid?utm_source=unsplash&amp;utm_medium=referral&amp;utm_content=creditCopyText\">National Institute of Allergy and Infectious Diseases<\/a> on <a href=\"https:\/\/unsplash.com\/photos\/a-person-in-a-lab-holding-a-pipe-Cdt3s_jp1OY?utm_source=unsplash&amp;utm_medium=referral&amp;utm_content=creditCopyText\">Unsplas<\/a>h<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Pasa\u017cowanie kom\u00f3rek jest rutynowo wykonywan\u0105 czynno\u015bci\u0105. Kom\u00f3rki rosn\u0105ce w zawiesinie nie stwarzaj\u0105 problem\u00f3w. Wyb\u00f3r odpowiedniej metody odklejania kom\u00f3rek od pod\u0142o\u017ca (tzw. dysocjacji) zale\u017cy od typu&hellip;<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":107,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_sitemap_exclude":false,"_sitemap_priority":"","_sitemap_frequency":"","footnotes":""},"class_list":["post-1811","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.6 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Metody odklejania kom\u00f3rek - Bioeducator.eu<\/title>\n<meta name=\"description\" content=\"Metody odklejania kom\u00f3rek od pod\u0142o\u017ca. W artykule opisujemy r\u00f3\u017cne techniki odklejania. 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