{"id":1004,"date":"2017-02-27T21:19:33","date_gmt":"2017-02-27T21:19:33","guid":{"rendered":"https:\/\/www.progeriaresearch.org\/?page_id=1004"},"modified":"2025-02-11T09:34:37","modified_gmt":"2025-02-11T14:34:37","slug":"lymphoblast-cell-culture-protocols","status":"publish","type":"page","link":"https:\/\/www.progeriaresearch.org\/it\/lymphoblast-cell-culture-protocols\/","title":{"rendered":"Protocolli di coltura cellulare linfoblastica"},"content":{"rendered":"<p>[et_pb_section fb_built=\u201d1\u2033 fullwidth=\u201don\u201d disabled_on=\u201doff|off|off\u201d _builder_version=\u201d4.16\u2033 border_width_bottom=\u201d55px\u201d border_color_bottom=\u201d#29327a\u201d locked=\u201doff\u201d global_colors_info=\u201d{}\u201d][et_pb_fullwidth_header _builder_version=\u201d4.16\u2033 title_font=\u201d||||||||\u201d title_font_size=\u201d55\u2033 background_color=\u201d#29327a\u201d background_image=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2020\/12\/Jes.jpg\u201d background_position=\u201dcenter_left\u201d custom_padding=\u201d9vw||9vw||true\u201d custom_padding_tablet=\u201d\u201d custom_padding_phone=\u201d|56px||\u201d custom_padding_last_edited=\u201don|desktop\u201d title_font_size_tablet=\u201d45px\u201d title_font_size_phone=\u201d40px\u201d title_font_size_last_edited=\u201don|phone\u201d z_index_tablet=\u201d500\u2033 custom_css_main_element=\u201dbackground-position: center 18% !important;\u201d global_colors_info=\u201d{}\u201d]<\/p>\n<h1 class=\"vc_custom_heading\" data-fontsize=\"34\" data-lineheight=\"48\" style=\"text-align: right;\">Cellula linfoblastica<\/h1>\n<h1 class=\"vc_custom_heading\" data-fontsize=\"34\" data-lineheight=\"48\" style=\"text-align: right;\">Protocolli di coltura<\/h1>\n<p>&nbsp;<\/p>\n<p>[\/et_pb_fullwidth_header][\/et_pb_section][et_pb_section fb_built=\u201d1\u2033 use_custom_gutter=\u201don\u201d gutter_width=\u201d1\u2033 specialty=\u201don\u201d padding_left_1=\u201d35px\u201d padding_left_2=\u201d35px\u201d padding_2_tablet=\u201d|||0px\u201d padding_2_phone=\u201d\u201d padding_2_last_edited=\u201don|desktop\u201d module_class_1=\u201dsidebar-secondary-nav\u201d module_class=\u201dhandprint-bg\u201d _builder_version=\u201d4.16\u2033 background_image=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2019\/04\/blue-handprint-only.png\u201d parallax=\u201don\u201d parallax_method=\u201doff\u201d larghezza_interna=\u201d100%\u201d larghezza_interna_massima=\u201d100%\u201d imbottitura_personalizzata=\u201d0|0px|54px|0px|falso|falso\u201d z_index_tablet=\u201d500\u2033 larghezza_interna_massima=\u201d10px\u201d colore_interna_massima=\u201d#8fd2ed\u201d usa_larghezza_personalizzata=\u201don\u201d unit\u00e0_di_larghezza=\u201doff\u201d percentuale_larghezza_personalizzata=\u201d100%\u201d info_colori_globali=\u201d{}\u201d][et_pb_column tipo=\u201d1_4\u2033 versione_builder=\u201d4.16\u2033 imbottitura_personalizzata=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_sidebar area=\u201det_pb_widget_area_14\u2033 disabled_on=\u201don|on|off\u201d module_class=\u201dsubpage-sidebars\u201d _builder_version=\u201d4.16\u2033 animation_style=\u201dfade\u201d z_index_tablet=\u201d500\u2033 border_width_right=\u201d5px\u201d locked=\u201doff\u201d global_colors_info=\u201d{}\u201d]<br \/>\n[\/et_pb_sidebar][\/et_pb_column][et_pb_column type=\u201d3_4\u2033 specialty_columns=\u201d3\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_row_inner _builder_version=\u201d4.16\u2033 background_color=\u201d#00b2e2\u2033 custom_margin=\u201d41px||\u201d custom_margin_tablet=\u201d0px||\u201d custom_margin_phone=\u201d\u201d custom_margin_last_edited=\u201don|phone\u201d custom_padding=\u201d39.4375px|20px|0|20px|false|true\u201d animation_style=\u201dslide\u201d animation_direction=\u201dtop\u201d animation_intensity_slide=\u201d25%\u201d box_shadow_style=\u201dpreset1\u2033 box_shadow_blur=\u201d38px\u201d box_shadow_spread=\u201d-12px\u201d locked=\u201doff\u201d global_colors_info=\u201d{}\u201d][et_pb_column_inner saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_text admin_label=\u201dIl lavoro della Progeria Research Foundation\u2026\u201d _builder_version=\u201d4.16\u2033 header_2_line_height=\u201d1.2em\u201d background_layout=\u201ddark\u201d custom_margin=\u201d||35px\u201d custom_padding=\u201d|50px||\u201d custom_padding_tablet=\u201d|50px||50px||true\u201d custom_padding_phone=\u201d\u201d custom_padding_last_edited=\u201don|desktop\u201d header_font_size_tablet=\u201d\u201d header_font_size_phone=\u201d\u201d header_font_size_last_edited=\u201don|desktop\u201d header_2_font_size_tablet=\u201d25px\u201d header_2_font_size_phone=\u201d\u201d header_2_font_size_last_edited=\u201don|tablet\u201d header_4_font_size_tablet=\u201d\u201d header_4_font_size_phone=\u201d19px\u201d header_4_font_size_last_edited=\u201don|phone\u201d header_4_line_height_tablet=\u201d\u201d header_4_line_height_phone=\u201d1.2em\u201d header_4_line_height_last_edited=\u201don|phone\u201d z_index_tablet=\u201d500\u2033 global_colors_info=\u201d{}\u201d]<\/p>\n<h2 style=\"text-align: center;\"><strong>Protocollo per la coltura dei linfociti trasformati<\/strong><\/h2>\n<p>[\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner custom_padding_last_edited=\u201don|phone\u201d _builder_version=\u201d4.16\u2033 custom_padding=\u201d|35px|0|0px|false|false\u201d custom_padding_tablet=\u201d|35px||35px||true\u201d custom_padding_phone=\u201d\u201d animation_direction=\u201dtop\u201d global_colors_info=\u201d{}\u201d][et_pb_column_inner saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_text _builder_version=\u201d4.27.4\u2033 background_size=\u201dinitial\u201d background_position=\u201dtop_left\u201d background_repeat=\u201drepeat\u201d custom_padding=\u201d||0px||false|false\u201d hover_enabled=\u201d0\u2033 global_colors_info=\u201d{}\u201d sticky_enabled=\u201d0\u2033]<\/p>\n<h4 style=\"text-align: center;\"><strong>Protocollo per la coltura dei linfoblasti<\/strong><\/h4>\n<h4><strong>Terreno di coltura<\/strong><\/h4>\n<p>Modello RPMI 1640 \u2013 ThermoFisher # 11875<br \/>15% Siero bovino fetale (FBS) \u2013 ThermoFisher #10437-028<br \/>1% (1X) Penicillina-Streptomicina (facoltativo) \u2013 ThermoFisher #15140-122<\/p>\n<h4><strong><\/strong><\/h4>\n<h4><strong>Note generali<br \/><\/strong><\/h4>\n<ul>\n<li>Il terreno di coltura deve essere sempre equilibrato nell&#039;incubatrice a 37\u00b0C, 5% CO<sub>2<\/sub> per 30 minuti prima dell&#039;uso<\/li>\n<li>Utilizzare sempre fiaschette con tappi ventilati<\/li>\n<li>Incubare sempre il pallone in posizione verticale<\/li>\n<li>Non utilizzare pi\u00f9 di 20 ml in una beuta T25<\/li>\n<\/ul>\n<h4><strong><\/strong><\/h4>\n<h4><strong>Scongelamento delle cellule congelate<br \/><\/strong><\/h4>\n<p>Le cellule congelate saranno ricevute in aliquote da 1 ml su ghiaccio secco. Mantenere le cellule congelate fino allo scongelamento per la coltura cellulare o conservarle in azoto liquido se non si avviano subito le colture. Tuttavia, si raccomanda che se si intende conservare le cellule per un uso successivo, si creino scorte fresche e si congelano pi\u00f9 fiale. Le cellule sono crioconservate in 45% RPMI-1640, 50% FBS e 5% DMSO (grado di coltura tissutale).<\/p>\n<ol>\n<li style=\"list-style-type: none;\">\n<ol>\n<li>Versare 10 ml di RPMI-1640, siero fetale bovino (FBS) 15%, \u00b1 antibiotici in una fiaschetta T-25 con tappo ventilato.<\/li>\n<li>Lasciare che il terreno si stabilizzi in un incubatore umidificato a 37\u00b0C con CO 5%<sub>2<\/sub> all&#039;aria per 30 minuti. Le beute devono essere incubate in posizione verticale.<\/li>\n<li>Scongelare ciascuna fiala o flacone di cellule congelate in un bagno d&#039;acqua a 37\u00b0C o in un becher di acqua tiepida, una alla volta.<\/li>\n<li>Pulisci rapidamente l&#039;esterno dell&#039;ampolla o della fiala con etanolo 70%. Se le cellule sono in un&#039;ampolla di vetro, romperla con attenzione, assicurandoti di proteggere le dita dal vetro rotto.<\/li>\n<li>Rimuovere 1 ml di cellule congelate con una pipetta sterile e metterle in una beuta T-25 con 10 ml di terreno. Etichettare chiaramente la beuta per identificare ogni linea cellulare.<\/li>\n<li>Posizionare il pallone in un incubatore umidificato a 37\u00b0C con 5% CO<sub>2<\/sub> nell&#039;aria. Le beute devono essere incubate in posizione verticale.<\/li>\n<li>Dopo 24 ore, rimuovere 5 ml di terreno dalla parte superiore del pallone, senza toccare i linfociti depositati sul fondo, e sostituirli con 5 ml di terreno preriscaldato.<\/li>\n<li>Procedere con il protocollo di coltura delle cellule linfoblastiche.<\/li>\n<\/ol>\n<\/li>\n<\/ol>\n<h4><strong><\/strong><\/h4>\n<h4><strong>Coltura linfoblastica<\/strong><\/h4>\n<ol>\n<li style=\"list-style-type: none;\">\n<ol>\n<li>Tre o quattro giorni dopo lo scongelamento, conta le cellule. Le cellule non sono aderenti e crescono in grumi. Rompere i grumi pipettando delicatamente.<\/li>\n<li>Contare le cellule.<\/li>\n<li>Idealmente, la concentrazione cellulare non dovrebbe superare 2 x 10<sup>6<\/sup> cellule\/ml.<\/li>\n<li>A seconda della concentrazione cellulare, reinserire, dividere o congelare le cellule.<\/li>\n<li>Rialimentare aggiungendo 5-6 ml di terreno di coltura equilibrato.<\/li>\n<li>Seminare nuove colture a non meno di 2 x 10<sup>5<\/sup> cellule\/ml.<\/li>\n<li>Le cellule devono essere congelate a circa 5 x 10<sup>6<\/sup> cellule\/ml. Seguire il protocollo di congelamento cellulare.<\/li>\n<\/ol>\n<\/li>\n<\/ol>\n<h4><strong><\/strong><\/h4>\n<h4><strong>Congelamento dei linfoblasti<\/strong><\/h4>\n<ol>\n<li style=\"list-style-type: none;\">\n<ol>\n<li>Le cellule devono essere congelate a circa 5 x 10<sup>6<\/sup> cellule\/ml.<\/li>\n<li>Trasferire il volume appropriato di cellule in una provetta conica da 15 o 50 ml.<\/li>\n<li>Centrifugare a 4\u00b0C a 100 xg per 10 minuti.<\/li>\n<li>Rimuovere il terreno e risospenderlo in un volume appropriato di terreno di congelamento freddo.<\/li>\n<li>Trasferire 1 ml di cellule ciascuna nella crioprovetta.<\/li>\n<li>Collocare le crioprovette nella camera di congelamento e riporre la camera nel congelatore a -80\u00b0C per tutta la notte.<\/li>\n<li>Il giorno successivo, trasferire le crioprovette nell&#039;azoto liquido.<\/li>\n<\/ol>\n<\/li>\n<\/ol>\n<p>[\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner _builder_version=\u201d4.27.4\u2033 _module_preset=\u201ddefault\u201d hover_enabled=\u201d0\u2033 global_colors_info=\u201d{}\u201d sticky_enabled=\u201d0\u2033 custom_padding=\u201d24px||24px|||\u201d][et_pb_column_inner saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.27.4\u2033 _module_preset=\u201ddefault\u201d global_colors_info=\u201d{}\u201d][et_pb_button button_url=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2025\/02\/Lymphoblast-protocol-1-22-25.pdf\u201d button_text=\u201dProtocollo per la coltura dei linfoblasti\u201d admin_label=\u201dProtocollo per la coltura dei linfoblasti\u201d _builder_version=\u201d4.27.4\u2033 _module_preset=\u201ddefault\u201d button_text_color=\u201d#FFFFFF\u201d button_bg_color=\u201d#00B2E2\u2033 button_border_color=\u201d#00B2E2\u2033 global_colors_info=\u201d{}\u201d][\/et_pb_button][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner custom_padding_last_edited=\u201don|phone\u201d _builder_version=\u201d4.16\u2033 custom_padding=\u201d||0px||||\u201d custom_padding_tablet=\u201d|35px||35px||true\u201d custom_padding_phone=\u201d\u201d border_width_top=\u201d10px\u201d border_color_top=\u201d#8fd2ed\u201d global_colors_info=\u201d{}\u201d][et_pb_column_inner saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_text _builder_version=\u201d4.16\u2033 global_colors_info=\u201d{}\u201d]<\/p>\n<h4><strong>Per ulteriori informazioni contattare:<\/strong><\/h4>\n<p>[\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner column_structure=\u201d1_2,1_2\u2033 custom_padding_last_edited=\u201don|phone\u201d _builder_version=\u201d4.16\u2033 custom_padding=\u201d|35px|0|0px|false|false\u201d custom_padding_tablet=\u201d|35px||35px||true\u201d custom_padding_phone=\u201d\u201d animation_direction=\u201dtop\u201d global_colors_info=\u201d{}\u201d][et_pb_column_inner type=\u201d1_2\u2033 saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|20px||\u201d custom_padding_tablet=\u201d||20px\u201d custom_padding_phone=\u201d\u201d custom_padding_last_edited=\u201don|desktop\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_text _builder_version=\u201d4.19.5\u2033 global_colors_info=\u201d{}\u201d]<\/p>\n<h6>Dott. Leslie B. Gordon, dottore in medicina e chirurgia<\/h6>\n<p>Professore di ricerca pediatrica Warren Alpert Medical School della Brown University e Dipartimento di pediatria, Hasbro Children&#039;s Hospital, Providence, RI Dipartimento di anestesia, Children&#039;s Hospital Boston e Harvard Medical School, Boston, MA Direttore medico, The Progeria Research Foundation<\/p>\n<p>Telefono: 978-535-2594<br \/>Fax: 508-543-0377<br \/><span> <\/span><a href=\"mailto:lgordon@progeriaresearch.org\" title=\"mailto:lgordon@progeriaresearch.org\">lgordon@progeriaresearch.org<\/a><\/p>\n<p>[\/et_pb_text][\/et_pb_column_inner][et_pb_column_inner type=\u201d1_2\u2033 saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||20px\u201d custom_padding_tablet=\u201d20px|||0px\u201d custom_padding_phone=\u201d\u201d custom_padding_last_edited=\u201don|phone\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_text _builder_version=\u201d4.27.2\u2033 custom_padding=\u201d||0px||\u201d custom_padding_last_edited=\u201doff|desktop\u201d global_colors_info=\u201d{}\u201d]<\/p>\n<h6><span>Wendy Norris<\/span><\/h6>\n<div><span>Banca di cellule e tessuti PRF<br \/>Telefono: 401-274-1122 x 48063<br \/><a href=\"mailto:wnorris@brownhealth.org\" title=\"mailto:wnorris@brownhealth.org\" data-outlook-id=\"6ae643af-3829-46f7-8b6a-2ef767034802\">e-mail: wnorris@brownhealth.org<\/a><br \/><\/span><\/div>\n<p>Italiano: [\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][\/et_pb_column][\/et_pb_section][et_pb_section fb_built=\u201d1\u2033 module_class=\u201dfooter\u201d _builder_version=\u201d4.21.0\u2033 background_color=\u201d#29327a\u201d custom_margin=\u201d-2px|||||\u201d custom_padding=\u201d0|0px|0|0px|false|false\u201d z_index_tablet=\u201d500\u2033 border_width_top=\u201d12px\u201d border_color_top=\u201d#00b2e2\u2033 global_module=\u201d133\u2033 locked=\u201doff\u201d global_colors_info=\u201d{}\u201d][et_pb_row_inner column_structure=\u201d1_3,1_3,1_3\u2033 disabled_on=\u201don|on|on\u201d admin_label=\u201dRow\u201d _builder_version=\u201d4.21.0\u2033 _module_preset=\u201ddefault\u201d min_height=\u201d277px\u201d custom_margin=\u201d|20px|30px||false|false\u201d custom_padding=\u201d0px|30px|0px||false|false\u201d disabled=\u201don\u201d global_colors_info=\u201d{}\u201d][et_pb_column_inner type=\u201d1_3\u2033 saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.21.0\u2033 _module_preset=\u201ddefault\u201d global_colors_info=\u201d{}\u201d][et_pb_image src=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2023\/08\/Startbanner-1.png\u201d title_text=\u201dStartbanner\u201d url=\u201dhttps:\/\/www.gravoc.com\/\u201d align=\u201dright\u201d _builder_version=\u201d4.21.0\u2033 _module_preset=\u201ddefault\u201d custom_margin=\u201d|-50px||20px|false|false\u201d global_colors_info=\u201d{}\u201d][\/et_pb_image][\/et_pb_column_inner][et_pb_column_inner tipo=\u201d1_3\u2033 saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.21.0\u2033 _module_preset=\u201ddefault\u201d global_colors_info=\u201d{}\u201d][\/et_pb_column_inner][et_pb_column_inner tipo=\u201d1_3\u2033 saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.21.0\u2033 _module_preset=\u201ddefault\u201d global_colors_info=\u201d{}\u201d][et_pb_image Italiano: src=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2023\/07\/RFRFinish.png\u201d title_text=\u201dRFRFinish\u201d _builder_version=\u201d4.21.0\u2033 _module_preset=\u201ddefault\u201d custom_margin=\u201d|20px||-50px|false|false\u201d global_colors_info=\u201d{}\u201d][\/et_pb_image][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row column_structure=\u201d1_4,1_4,1_2\u2033 make_equal=\u201don\u201d module_class=\u201d et_pb_row_fullwidth\u201d _builder_version=\u201d4.16\u2033 width=\u201d89%\u201d width_tablet=\u201d80%\u201d width_phone=\u201d\u201d larghezza_ultima_modifica=\u201dsu|desktop\u201d larghezza_massima=\u201d89%\u201d larghezza_massima_tablet=\u201d80%\u201d larghezza_massima_telefono=\u201d\u201d larghezza_massima_modifica=\u201dsu|desktop\u201d z_index_tablet=\u201d500\u2033 make_fullwidth=\u201dsu\u201d larghezza_unit\u00e0=\u201dspento\u201d percentuale_larghezza_personalizzata=\u201d100%\u201d info_colori_globali=\u201d{}\u201d][et_pb_column tipo=\u201d1_4\u2033 versione_builder=\u201d4.16\u2033 riempimento_personalizzato=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_cta button_url=\u201dhttps:\/\/lp.constantcontactpages.com\/sl\/88gWWwz\u201d button_text=\u201dRegistrati ora\u201d admin_label=\u201dRegistrati per gli aggiornamenti\u201d module_class=\u201dsign-btn\u201d _builder_version=\u201d4.27.4\u2033 header_font_size=\u201d25px\u201d background_color=\u201d#29327a\u201d animation_style=\u201dslide\u201d animation_direction=\u201dleft\u201d animation_intensity_slide=\u201d25%\u201d link_option_url=\u201dhttps:\/\/lp.constantcontactpages.com\/sl\/88gWWwz\u201d header_font_size_tablet=\u201d\u201d header_font_size_phone=\u201d30px\u201d header_font_size_last_edited=&quot;on|desktop&quot; z_index_tablet=&quot;500&quot; border_radii=&quot;on|25px|25px|25px|25px&quot; global_colors_info=&quot;{}&quot; button_bg_color__hover_enabled=&quot;on&quot; button_bg_color__hover=&quot;#8fd2ed&quot; button_border_color__hover_enabled=&quot;on&quot;]<\/p>\n<h2>Iscrizione<\/h2>\n<h2>per il nostro<\/h2>\n<h2>Aggiornamenti!<\/h2>\n<p>[\/et_pb_cta][\/et_pb_column][et_pb_column type=\u201d1_4\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_cta button_url=\u201dhttps:\/\/progeriaresearch.donorsupport.co\/-\/XZHJVWZR\u201d button_text=\u201dFai una donazione ora\u201d admin_label=\u201dInsieme troveremo la cura!\u201d module_class=\u201dsign-btn\u201d _builder_version=\u201d4.16\u2033 header_font_size=\u201d25px\u201d background_color=\u201d#29327a\u201d animation_style=\u201dslide\u201d animation_direction=\u201dleft\u201d animation_intensity_slide=\u201d25%\u201d header_font_size_tablet=\u201d\u201d header_font_size_phone=\u201d30px\u201d header_font_size_last_edited=\u201don|desktop\u201d body_font_size_tablet=\u201d\u201d body_font_size_phone=\u201d\u201d body_font_size_last_edited=\u201don|desktop\u201d z_index_tablet=\u201d500\u2033 border_radii=\u201don|25px|25px|25px|25px\u201d global_colors_info=\u201d{}\u201d button_bg_color__hover_enabled=\u201don\u201d colore_bg_pulsante__hover=\u201d#8fd2ed\u201d colore_bordo_pulsante__hover_abilitato=\u201don\u201d]<\/p>\n<h2>Insieme, noi<\/h2>\n<h2><em>VOLERE<\/em><\/h2>\n<h2>trova la cura!<\/h2>\n<p>[\/et_pb_cta][\/et_pb_column][et_pb_column tipo=\u201d1_2\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_image src=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2024\/08\/2024-strip-footer-strip-copy.png\u201d title_text=\u201d2024 strip footer strip copy\u201d _builder_version=\u201d4.27.0\u2033 _module_preset=\u201ddefault\u201d custom_margin=\u201d35px||||false|false\u201d global_colors_info=\u201d{}\u201d][\/et_pb_image][\/et_pb_column][\/et_pb_row][\/et_pb_section]<\/p>","protected":false},"excerpt":{"rendered":"<p>[et_pb_section fb_built=\u201d1\u2033 fullwidth=\u201don\u201d disabled_on=\u201doff|off|off\u201d _builder_version=\u201d4.16\u2033 border_width_bottom=\u201d55px\u201d border_color_bottom=\u201d#29327a\u201d locked=\u201doff\u201d global_colors_info=\u201d{}\u201d][et_pb_fullwidth_header _builder_version=\u201d4.16\u2033 title_font=\u201d||||||||\u201d title_font_size=\u201d55\u2033 background_color=\u201d#29327a\u201d background_image=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2020\/12\/Jes.jpg\u201d background_position=\u201dcenter_left\u201d custom_padding=\u201d9vw||9vw||true\u201d custom_padding_tablet=\u201d\u201d custom_padding_phone=\u201d|56px||\u201d custom_padding_last_edited=\u201don|desktop\u201d title_font_size_tablet=\u201d45px\u201d title_font_size_phone=\u201d40px\u201d title_font_size_last_edited=\u201don|phone\u201d z_index_tablet=\u201d500\u2033 custom_css_main_element=\u201dbackground-position: center 18% !important;\u201d global_colors_info=\u201d{}\u201d] Protocolli di coltura cellulare linfoblastica [\/et_pb_fullwidth_header][\/et_pb_section][et_pb_section fb_built=\u201d1\u2033 use_custom_gutter=\u201don\u201d gutter_width=\u201d1\u2033 specialty=\u201don\u201d padding_left_1=\u201d35px\u201d padding_left_2=\u201d35px\u201d padding_2_tablet=\u201d|||0px\u201d padding_2_phone=\u201d\u201d padding_2_last_edited=\u201don|desktop\u201d module_class_1=\u201dsidebar-secondary-nav\u201d module_class=\u201dhandprint-bg\u201d _builder_version=\u201d4.16\u2033 background_image=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2019\/04\/blue-handprint-only.png\u201d parallax=\u201don\u201d parallax_method=\u201doff\u201d inner_width=\u201d100%\u201d inner_max_width=\u201d100%\u201d custom_padding=\u201d0|0px|54px|0px|false|false\u201d z_index_tablet=\u201d500\u2033 border_width_top=\u201d10px\u201d border_color_top=\u201d#8fd2ed\u201d [\u2026]<\/p>","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_et_pb_use_builder":"on","_et_pb_old_content":"\t\t\t\t[vc_row][vc_column][vc_custom_heading text=\"Lymphoblast Cell Culture Protocols\" font_container=\"tag:h1|text_align:center\" use_theme_fonts=\"yes\"][vc_column_text]<strong>Protocol for Culturing Transformed Lymphocytes<\/strong>\r\n\r\nFrom Shipment of Frozen Cells\r\n\r\nFrozen cells will be received in 1 ml aliquots on dry ice. Keep cells frozen until thawed for cell culture or place in liquid nitrogen storage if not starting cultures right away. However, it is recommended that if you are going to store the cells for later use, grow up fresh stocks and freeze multiple ampoules. The cells are cryopreserved in 45% RPMI-1640, 50% fetal calf serum, and 5% DMSO (tissue culture grade).\r\n<ol>\r\n \t<li>Place 10 ml of RPMI-1640, 15% fetal calf serum (FCS), \u00b1antibiotics in a T-25 flask.<\/li>\r\n \t<li>Allow the media to equilibrate in a 37\u00b0C humidified incubator with 5% CO 2in air for 30 minutes.<\/li>\r\n \t<li>Thaw each ampoule of frozen cells in a 37\u00b0C water bath or a beaker of lukewarm water, one at a time.<\/li>\r\n \t<li>Quickly clean the outside of the ampoule off with a sterile isopropanol prep pad, then wrap the prep pad around the ampoule to protect fingers and crack the seal on the ampoule inside a biological safety hood.<\/li>\r\n \t<li>Remove the 1 ml of frozen cells with a sterile glass Pasteur pipet and place in a T-25 flask with 10 ml of medium. Clearly label flask to identify each cell line.<\/li>\r\n \t<li>Place the cells in a 37\u00b0C humidified incubator with 5% CO 2in the air.<\/li>\r\n \t<li>After 24 hours remove 5 ml of medium from the top without disturbing lymphocytes settled on the bottom of the flask and replace with 5 ml of pre-warmed medium.<\/li>\r\n \t<li>Feed the cells with 5 - 6 ml of fresh medium every 3 - 4 days and split into new cultures as required.<\/li>\r\n<\/ol>\r\n<strong>From Shipment of Live Cultures<\/strong>\r\n<ol>\r\n \t<li>Live lymphocyte cultures will be received in T-25 flasks filled to the top with RPMI-1640, 15% FCS, 1% Penicillin-Streptomycin (Gibco). The caps will be tight and sealed with parafilm.<\/li>\r\n \t<li>Remove parafilm.<\/li>\r\n \t<li>Place cultures in a 37\u00b0C humidified incubator with 5% CO2 in air and allow cells to settle out (20 - 30 minutes).<\/li>\r\n \t<li>Using a sterile pipet remove all but 10 - 15 ml of medium and discard.<\/li>\r\n \t<li>Place caps back on flasks loosened slightly to allow gas exchange (Check to make sure cap threads do not have media on them. If so you may want to transfer to a new flask.) Place the cultures back in the incubator with 5% CO 2in air. They should recover in a day or two.<\/li>\r\n \t<li>Feed the cells with fresh medium every 3 - 4 days by resuspending the cell aggregates and removing 50 -60% of the volume and replacing with fresh media. Cells removed from the flask can be used to initiate new cultures as needed.<\/li>\r\n<\/ol>\r\n<strong>For Further Information Please Contact:<\/strong>\r\n\r\nLeslie B. Gordon, MD, PhD\r\nProfessor of Pediatrics Research\u00a0Warren Alpert Medical School of Brown University and Department of Pediatrics, Hasbro Children's Hospital, Providence, RI\u00a0Department of Anesthesia, Children's Hospital Boston and Harvard Medical School, Boston, MA\u00a0Medical Director, The Progeria Research Foundation\r\n\r\nPhone: 978-535-2594\r\nFax: 508-543-0377\r\n<a href=\"mailto:Leslie_Gordon@brown.edu\">Leslie_Gordon@brown.edu<\/a>\r\n<div><span style=\"font-family: Georgia, Palatino;\">Wendy Norris<\/span><\/div>\r\n<div><span style=\"font-family: Georgia, Palatino;\">PRF Cell and Tissue Bank\r\nPhone: 401-274-1122 x 48063\r\n<a href=\"mailto:wnorris@lifespan.org\" target=\"_blank\" rel=\"noopener noreferrer\">wnorris@lifespan.org<\/a><\/span><\/div>\r\n[\/vc_column_text][\/vc_column][\/vc_row]\t\t","_et_gb_content_width":"","footnotes":"","_links_to":"","_links_to_target":""},"class_list":["post-1004","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v26.8 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Lymphoblast | The Progeria Research Foundation<\/title>\n<meta name=\"description\" content=\"Lymphoblast Cell Culture Protocols are listed within our website, regarding from shipment of frozen cells and live cultures.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.progeriaresearch.org\/it\/lymphoblast-cell-culture-protocols\/\" \/>\n<meta property=\"og:locale\" content=\"it_IT\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Lymphoblast | The Progeria Research Foundation\" \/>\n<meta property=\"og:description\" content=\"Lymphoblast Cell Culture Protocols are listed within our website, regarding from shipment of frozen cells and live cultures.\" \/>\n<meta property=\"og:url\" content=\"https:\/\/www.progeriaresearch.org\/it\/lymphoblast-cell-culture-protocols\/\" \/>\n<meta property=\"og:site_name\" content=\"The Progeria Research Foundation\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/ProgeriaResearch\/\" \/>\n<meta property=\"article:modified_time\" content=\"2025-02-11T14:34:37+00:00\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:site\" content=\"@Progeria\" \/>\n<meta name=\"twitter:label1\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data1\" content=\"4 minuti\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"WebPage\",\"@id\":\"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/\",\"url\":\"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/\",\"name\":\"Lymphoblast | The Progeria Research Foundation\",\"isPartOf\":{\"@id\":\"https:\/\/www.progeriaresearch.org\/ta\/#website\"},\"datePublished\":\"2017-02-27T21:19:33+00:00\",\"dateModified\":\"2025-02-11T14:34:37+00:00\",\"description\":\"Lymphoblast Cell Culture Protocols are listed within our website, regarding from shipment of frozen cells and live cultures.\",\"breadcrumb\":{\"@id\":\"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/#breadcrumb\"},\"inLanguage\":\"it-IT\",\"potentialAction\":[{\"@type\":\"ReadAction\",\"target\":[\"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/\"]}]},{\"@type\":\"BreadcrumbList\",\"@id\":\"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/#breadcrumb\",\"itemListElement\":[{\"@type\":\"ListItem\",\"position\":1,\"name\":\"Home\",\"item\":\"https:\/\/www.progeriaresearch.org\/\"},{\"@type\":\"ListItem\",\"position\":2,\"name\":\"Lymphoblast Cell Culture Protocols\"}]},{\"@type\":\"WebSite\",\"@id\":\"https:\/\/www.progeriaresearch.org\/ta\/#website\",\"url\":\"https:\/\/www.progeriaresearch.org\/ta\/\",\"name\":\"The Progeria Research Foundation\",\"description\":\"For the Children \u2665 For the Cure\",\"publisher\":{\"@id\":\"https:\/\/www.progeriaresearch.org\/ta\/#organization\"},\"potentialAction\":[{\"@type\":\"SearchAction\",\"target\":{\"@type\":\"EntryPoint\",\"urlTemplate\":\"https:\/\/www.progeriaresearch.org\/ta\/?s={search_term_string}\"},\"query-input\":{\"@type\":\"PropertyValueSpecification\",\"valueRequired\":true,\"valueName\":\"search_term_string\"}}],\"inLanguage\":\"it-IT\"},{\"@type\":\"Organization\",\"@id\":\"https:\/\/www.progeriaresearch.org\/ta\/#organization\",\"name\":\"The Progeria Research Foundation\",\"url\":\"https:\/\/www.progeriaresearch.org\/ta\/\",\"logo\":{\"@type\":\"ImageObject\",\"inLanguage\":\"it-IT\",\"@id\":\"https:\/\/www.progeriaresearch.org\/ta\/#\/schema\/logo\/image\/\",\"url\":\"https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2019\/05\/PRF_Logo_2019_optimized.png\",\"contentUrl\":\"https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2019\/05\/PRF_Logo_2019_optimized.png\",\"width\":300,\"height\":86,\"caption\":\"The Progeria Research Foundation\"},\"image\":{\"@id\":\"https:\/\/www.progeriaresearch.org\/ta\/#\/schema\/logo\/image\/\"},\"sameAs\":[\"https:\/\/www.facebook.com\/ProgeriaResearch\/\",\"https:\/\/x.com\/Progeria\",\"https:\/\/www.instagram.com\/progeriaresearch\/\"]}]}<\/script>\n<!-- \/ Yoast SEO plugin. -->","yoast_head_json":{"title":"Linfoblasto | Fondazione per la ricerca sulla progeria","description":"I protocolli per la coltura cellulare dei linfoblasti sono elencati sul nostro sito web, a partire dalla spedizione di cellule congelate e colture vive.","robots":{"index":"index","follow":"follow","max-snippet":"max-snippet:-1","max-image-preview":"max-image-preview:large","max-video-preview":"max-video-preview:-1"},"canonical":"https:\/\/www.progeriaresearch.org\/it\/lymphoblast-cell-culture-protocols\/","og_locale":"it_IT","og_type":"article","og_title":"Lymphoblast | The Progeria Research Foundation","og_description":"Lymphoblast Cell Culture Protocols are listed within our website, regarding from shipment of frozen cells and live cultures.","og_url":"https:\/\/www.progeriaresearch.org\/it\/lymphoblast-cell-culture-protocols\/","og_site_name":"The Progeria Research Foundation","article_publisher":"https:\/\/www.facebook.com\/ProgeriaResearch\/","article_modified_time":"2025-02-11T14:34:37+00:00","twitter_card":"summary_large_image","twitter_site":"@Progeria","twitter_misc":{"Est. reading time":"4 minuti"},"schema":{"@context":"https:\/\/schema.org","@graph":[{"@type":"WebPage","@id":"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/","url":"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/","name":"Linfoblasto | Fondazione per la ricerca sulla progeria","isPartOf":{"@id":"https:\/\/www.progeriaresearch.org\/ta\/#website"},"datePublished":"2017-02-27T21:19:33+00:00","dateModified":"2025-02-11T14:34:37+00:00","description":"I protocolli per la coltura cellulare dei linfoblasti sono elencati sul nostro sito web, a partire dalla spedizione di cellule congelate e colture vive.","breadcrumb":{"@id":"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/#breadcrumb"},"inLanguage":"it-IT","potentialAction":[{"@type":"ReadAction","target":["https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/"]}]},{"@type":"BreadcrumbList","@id":"https:\/\/www.progeriaresearch.org\/ru\/lymphoblast-cell-culture-protocols\/#breadcrumb","itemListElement":[{"@type":"ListItem","position":1,"name":"Home","item":"https:\/\/www.progeriaresearch.org\/"},{"@type":"ListItem","position":2,"name":"Lymphoblast Cell Culture Protocols"}]},{"@type":"WebSite","@id":"https:\/\/www.progeriaresearch.org\/ta\/#website","url":"https:\/\/www.progeriaresearch.org\/ta\/","name":"Fondazione per la ricerca sulla progeria","description":"Per i bambini \u2665 Per la cura","publisher":{"@id":"https:\/\/www.progeriaresearch.org\/ta\/#organization"},"potentialAction":[{"@type":"SearchAction","target":{"@type":"EntryPoint","urlTemplate":"https:\/\/www.progeriaresearch.org\/ta\/?s={search_term_string}"},"query-input":{"@type":"PropertyValueSpecification","valueRequired":true,"valueName":"search_term_string"}}],"inLanguage":"it-IT"},{"@type":"Organization","@id":"https:\/\/www.progeriaresearch.org\/ta\/#organization","name":"Fondazione per la ricerca sulla progeria","url":"https:\/\/www.progeriaresearch.org\/ta\/","logo":{"@type":"ImageObject","inLanguage":"it-IT","@id":"https:\/\/www.progeriaresearch.org\/ta\/#\/schema\/logo\/image\/","url":"https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2019\/05\/PRF_Logo_2019_optimized.png","contentUrl":"https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2019\/05\/PRF_Logo_2019_optimized.png","width":300,"height":86,"caption":"The Progeria Research Foundation"},"image":{"@id":"https:\/\/www.progeriaresearch.org\/ta\/#\/schema\/logo\/image\/"},"sameAs":["https:\/\/www.facebook.com\/ProgeriaResearch\/","https:\/\/x.com\/Progeria","https:\/\/www.instagram.com\/progeriaresearch\/"]}]}},"_links":{"self":[{"href":"https:\/\/www.progeriaresearch.org\/it\/wp-json\/wp\/v2\/pages\/1004","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.progeriaresearch.org\/it\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.progeriaresearch.org\/it\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.progeriaresearch.org\/it\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.progeriaresearch.org\/it\/wp-json\/wp\/v2\/comments?post=1004"}],"version-history":[{"count":0,"href":"https:\/\/www.progeriaresearch.org\/it\/wp-json\/wp\/v2\/pages\/1004\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.progeriaresearch.org\/it\/wp-json\/wp\/v2\/media?parent=1004"}],"curies":[{"name":"parola chiave","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}