{"id":1801,"date":"2026-01-07T12:27:41","date_gmt":"2026-01-07T11:27:41","guid":{"rendered":"https:\/\/bioeducator.eu\/?page_id=1801"},"modified":"2026-01-09T12:54:08","modified_gmt":"2026-01-09T11:54:08","slug":"pasazowanie-komorek","status":"publish","type":"page","link":"https:\/\/bioeducator.eu\/?page_id=1801","title":{"rendered":"Pasa\u017cowanie 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\">Kiedy hodujesz kom\u00f3rki by przeprowadzi\u0107 eksperymeny lub w jakimkolwiek innym celu, pierwszym etapem jest zazwyczaj ich namno\u017cenie by uzyska\u0107 ich odpowiedni\u0105 ilo\u015b\u0107. Nie mo\u017cna jednak ich hodowa\u0107 w niesko\u0144czono\u015b\u0107 w jednym naczyniu. Pr\u0119dzej czy p\u00f3\u017aniej sko\u0144czy im si\u0119 po prostu miejsce do \u017cycia. Zanim do tego dojdzie nale\u017cy wykona\u0107 pasa\u017cowanie kom\u00f3rek.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Pasa\u017cowanie kom\u00f3rek, zwane jest r\u00f3wnie\u017c subkultywacj\u0105. Jest to fundament pracy z liniami kom\u00f3rkowymi. Proces ten polega na przeniesieniu cz\u0119\u015bci kom\u00f3rek z dotychczasowego naczynia hodowlanego do nowego, ze \u015bwie\u017c\u0105 po\u017cywk\u0105. Cho\u0107 technicznie rutynowy, wymaga precyzji i zrozumienia fizjologii kom\u00f3rki, aby zapewni\u0107 powtarzalno\u015b\u0107 wynik\u00f3w eksperymentalnych.<\/p>\n<\/div>\n\n\n\n<div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow\"><div class=\"wp-block-image\">\n<figure class=\"aligncenter 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\" \/><figcaption class=\"wp-element-caption\">Pasa\u017cowanie kom\u00f3rek<\/figcaption><\/figure>\n<\/div><\/div>\n<\/div>\n\n\n\n<h2 class=\"wp-block-heading\">Po co pasa\u017cowa\u0107 kom\u00f3rki?<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Kom\u00f3rki w hodowli statycznej dziel\u0105 si\u0119, dop\u00f3ki pozwalaj\u0105 na to zasoby \u015brodowiska. Tymi zasobami s\u0105 sk\u0142adniki od\u017cywcze i miejsce do \u017cycia. W przypadku kom\u00f3rek rosn\u0105cych w zawiesinie jest to przestrze\u0144, w przypadku kom\u00f3rek adherentnych \u2013 powierzchnia. G\u0142\u00f3wnymi powodami, dla kt\u00f3rych musimy regularnie przeprowadza\u0107 pasa\u017cowanie, s\u0105:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Zapobieganie wyczerpaniu sk\u0142adnik\u00f3w od\u017cywczych:<\/strong> Glukoza, aminokwasy i czynniki wzrostu zawarte w surowicy ulegaj\u0105 zu\u017cyciu.<\/li>\n\n\n\n<li><strong>Kontrola pH:<\/strong> Produkty metabolizmu (np. kwas mlekowy) zakwaszaj\u0105 po\u017cywk\u0119, co mo\u017ce by\u0107 toksyczne dla kom\u00f3rek.<\/li>\n\n\n\n<li><strong>Unikanie hamowania kontaktowego:<\/strong> Wi\u0119kszo\u015b\u0107 kom\u00f3rek adherentnych przestaje si\u0119 dzieli\u0107 po osi\u0105gni\u0119ciu pe\u0142nej konfluencji (pokryciu ca\u0142ej powierzchni dna). Pozostawienie ich w tym stanie prowadzi do starzenia si\u0119 kom\u00f3rek (senescencji) i apoptozy.<\/li>\n\n\n\n<li><strong>Utrzymanie fazy logarytmicznej:<\/strong> Chcemy, aby kom\u00f3rki by\u0142y w fazie intensywnego wzrostu, co gwarantuje ich optymaln\u0105 kondycj\u0119 do do\u015bwiadcze\u0144.<\/li>\n<\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Pasa\u017cowanie kom\u00f3rek adherentnych vs. zawiesinowych<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">W zale\u017cno\u015bci od sposobu wzrostu kom\u00f3rek, procedura pasa\u017cowania znacz\u0105co si\u0119 r\u00f3\u017cni:<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Kom\u00f3rki adherentne (przylegaj\u0105ce)<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Wymagaj\u0105 powierzchni do wzrostu. Aby je pasa\u017cowa\u0107, nale\u017cy najpierw enzymatycznie lub mechanicznie zerwa\u0107 wi\u0105zania bia\u0142kowe \u0142\u0105cz\u0105ce kom\u00f3rk\u0119 z dnem naczynia oraz z s\u0105siednimi kom\u00f3rkami. Proces ten jest bardziej inwazyjny. Pasa\u017cowanie kom\u00f3rek adherentnych jest dok\u0142adnie om\u00f3wione w <a href=\"https:\/\/bioeducator.eu\/?page_id=1811\">innym artykule<\/a>.<\/p>\n\n\n<div class=\"wp-block-image\">\n<figure class=\"aligncenter 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-7gMpw5uj3Rk-unsplash-1024x683.jpg\" alt=\"\" class=\"wp-image-1803\" srcset=\"https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-7gMpw5uj3Rk-unsplash-1024x683.jpg 1024w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-7gMpw5uj3Rk-unsplash-300x200.jpg 300w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-7gMpw5uj3Rk-unsplash-768x512.jpg 768w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-7gMpw5uj3Rk-unsplash-391x260.jpg 391w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-7gMpw5uj3Rk-unsplash-1536x1025.jpg 1536w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/national-institute-of-allergy-and-infectious-diseases-7gMpw5uj3Rk-unsplash-2048x1366.jpg 2048w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n<\/div>\n\n\n<p class=\"wp-block-paragraph\"><strong>Kom\u00f3rki w zawiesinie<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Rosn\u0105 swobodnie w obj\u0119to\u015bci po\u017cywki (np. limfocyty, linie bia\u0142aczkowe). Pasa\u017cowanie jest tu znacznie prostsze i szybsze \u2013 polega na pobraniu cz\u0119\u015bci zawiesiny i przeniesieniu jej do nowego naczynia i \u015bwie\u017cego medium. Pasa\u017cu mo\u017cna dokona\u0107 np. poprzez rozcie\u0144czenie hodowli \u015bwie\u017cym medium i rozdzielenie jej do nowych naczy\u0144 lub przeniesieniu cz\u0119\u015bci lub ca\u0142o\u015bci do naczynia o wi\u0119kszej obj\u0119to\u015bci.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Rozcie\u0144czenie i jego wp\u0142yw na wzrost<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Po uzyskaniu zawiesiny kom\u00f3rek, musimy zdecydowa\u0107, w jakim stosunku (split ratio) przenie\u015b\u0107 je do nowych naczy\u0144 (np. 1:3, 1:4, 1:5, 1:10).<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Jak rozcie\u0144czenie wp\u0142ywa na kultur\u0119 in vitro?<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Zbyt niskie rozcie\u0144czenie (np. 1:2):<\/strong> Kom\u00f3rki bardzo szybko osi\u0105gn\u0105 konfluencj\u0119, co wymusza cz\u0119ste pasa\u017cowanie. <\/li>\n\n\n\n<li><strong>Zbyt wysokie rozcie\u0144czenie (np. 1:20):<\/strong> Mo\u017ce doprowadzi\u0107 do \u015bmierci kom\u00f3rek. Kom\u00f3rki komunikuj\u0105 si\u0119 ze sob\u0105 poprzez czynniki parakrynne. Je\u015bli odleg\u0142o\u015b\u0107 mi\u0119dzy nimi jest zbyt du\u017ca, nie osi\u0105gn\u0105 one &#8222;masy krytycznej&#8221; niezb\u0119dnej do stymulacji podzia\u0142\u00f3w.<\/li>\n\n\n\n<li><strong>G\u0119sto\u015b\u0107 posiewu:<\/strong> Dla ka\u017cdej linii kom\u00f3rkowej istnieje optymalna liczba kom\u00f3rek\/cm2 powierzchni, kt\u00f3ra zapewnia najkr\u00f3tszy czas podwojenia.<\/li>\n<\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Wp\u0142yw pasa\u017cu kom\u00f3rek na ich w\u0142a\u015bciwo\u015bci<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Numer pasa\u017cu jest niezwykle wa\u017cn\u0105 informacj\u0105 dla osoby prowadz\u0105cej hodowle in vitro. W miar\u0119 kolejnych pasa\u017cy linia kom\u00f3rkowa (nawet ta unie\u015bmiertelniona) mo\u017ce zmienia\u0107 swoje w\u0142a\u015bciwo\u015bci. Dzieje si\u0119 tak na skutek r\u00f3\u017cnych zjawisk, kt\u00f3re omawiam poni\u017cej<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Dryf genetyczny<\/strong> \u2013 Z ka\u017cdym kolejnym pasa\u017cem mog\u0105 kumulowa\u0107 si\u0119 mutacje punktowe i kariotypowe w kom\u00f3rkach.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Selekcja populacji<\/strong> \u2013 W hodowli in vitro poszczeg\u00f3lne kom\u00f3rki konkuruj\u0105 ze sob\u0105 o zasoby. Kom\u00f3rki dziel\u0105ce si\u0119 szybciej lub te, kt\u00f3re lepiej znosz\u0105 trypsynizacj\u0119 z czasem zaczynaj\u0105 dominowa\u0107 a r\u00f3\u017cnorodno\u015b\u0107 hodowli si\u0119 zmniejsza.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Zmiana ekspresji gen\u00f3w<\/strong> \u2013 wysoka liczba pasa\u017cy cz\u0119sto prowadzi do zmiany w profilu transkrypcyjnym. W liniach o wysokim pasa\u017cu (&gt;40) mo\u017ce dochodzi\u0107 do r\u00f3\u017cnicy w ekspresji nawet 1000 gen\u00f3w w por\u00f3wnaniu z liniami o pasa\u017cu niskim (15-20)<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Zmiany w morfologii<\/strong> \u2013 w niekt\u00f3rych liniach obserwuje si\u0119 zmiany kszta\u0142tu kom\u00f3rek w wysokich pasa\u017cach<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Zmiany w tempie wzrostu<\/strong> \u2013 w starych liniach, kom\u00f3rki dziel\u0105 si\u0119 zazwyczaj szybciej<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Utrata specjalizacji<\/strong> \u2013 wraz z wiekiem, zmianami genetycznymi i zmianami w ekspresji kom\u00f3rki mog\u0105 traci\u0107 swoje wyspecjalizowane funkcje. Przyk\u0142adowo kom\u00f3rki nowotworowe mog\u0105 nie reagowa\u0107 na hormony lub przesta\u0107 wytwarza\u0107 niekt\u00f3re bia\u0142ka<\/p>\n\n\n\n<h3 class=\"wp-block-heading\"><strong>Wp\u0142yw pasa\u017cu na wyniki bada\u0144<\/strong><\/h3>\n\n\n\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\">Jak wida\u0107 wp\u0142yw pasa\u017cowanie nie jest oboj\u0119tne dla linii kom\u00f3rkowej. By zachowa\u0107 por\u00f3wnywalno\u015b\u0107 wynik\u00f3w nale\u017cy zminimalizowa\u0107 ilo\u015b\u0107 pasa\u017cy wykonywanych w laboratorium. Zazwyczaj ustala si\u0119 zakres pasa\u017cy, w kt\u00f3rych mo\u017cna przeprowadza\u0107 dane badanie bez negatywnego wp\u0142ywu na jego wynik (np. podczas walidacji metody). Dobrym punktem startowym jest utrzymanie hodowli w zakresie 10 pasa\u017cy. By by\u0142o to mo\u017cliwe laboratorium powinno mie\u0107 odpowiednio du\u017cy bank zamro\u017conych kom\u00f3rek, z kt\u00f3rych mo\u017ce rozpoczyna\u0107 kolejne hodowle.<\/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-full\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"1024\" src=\"https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/consistency.png\" alt=\"consistency\" class=\"wp-image-1806\" srcset=\"https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/consistency.png 1024w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/consistency-300x300.png 300w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/consistency-150x150.png 150w, https:\/\/bioeducator.eu\/wp-content\/uploads\/2026\/01\/consistency-768x768.png 768w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n<\/div>\n<\/div>\n\n\n\n<p class=\"wp-block-paragraph\"><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Podsumowanie<\/strong><\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Pasa\u017cowanie to co\u015b wi\u0119cej ni\u017c tylko &#8222;przelewanie&#8221; kom\u00f3rek. To precyzyjne zarz\u0105dzanie cyklem \u017cyciowym populacji in vitro. Zrozumienie roli p\u0142ukania, kinetyki trypsynizacji oraz dynamiki wzrostu po rozcie\u0144czeniu pozwala na prowadzenie zdrowych i stabilnych genetycznie hodowli.<\/p>\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><a href=\"https:\/\/www.cellosaurus.org\/\">cellosaurus.org<\/a><\/li>\n\n\n\n<li><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/18072586\/\"><strong>Hughes et al. (2007):<\/strong> <em>&#8222;The costs of using unauthenticated, over-passaged cell lines: how much more data do we need?&#8221;<\/em><\/a><\/li>\n\n\n\n<li><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/25117809\/\"><strong>Geraghty et al. (2014):<\/strong> <em>&#8222;Guidelines for the use of cell lines in biomedical research&#8221;<\/em>.<\/a><\/li>\n\n\n\n<li>Hodowla kom\u00f3rek i tkanek in vitro. P.red. Stanis\u0142awy Stok\u0142osowej; 2004<\/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\">Unsplash<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Kiedy hodujesz kom\u00f3rki by przeprowadzi\u0107 eksperymeny lub w jakimkolwiek innym celu, pierwszym etapem jest zazwyczaj ich namno\u017cenie by uzyska\u0107 ich odpowiedni\u0105 ilo\u015b\u0107. Nie mo\u017cna jednak&hellip;<\/p>\n","protected":false},"author":1,"featured_media":1804,"parent":107,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_sitemap_exclude":false,"_sitemap_priority":"","_sitemap_frequency":"","footnotes":""},"class_list":["post-1801","page","type-page","status-publish","has-post-thumbnail","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.6 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Pasa\u017cowanie kom\u00f3rek - Bioeducator.eu<\/title>\n<meta name=\"description\" content=\"Pasa\u017cowanie kom\u00f3rek to podstawa ka\u017cdej udanej hodowli in vitro. 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