{"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\/ur\/lymphoblast-cell-culture-protocols\/","title":{"rendered":"\u0644\u06cc\u0645\u0641\u0648\u0628\u0644\u0627\u0633\u0679 \u0633\u06cc\u0644 \u06a9\u0644\u0686\u0631 \u067e\u0631\u0648\u0679\u0648\u06a9\u0648\u0644"},"content":{"rendered":"<p>[et_pb_section fb_built=\u201d1\u2033 full width=\u201don\u201d disabled_on=\u201doff|off|off\u201d _builder_version=\u201d4.16\u2033 border_width_bottom=\u201d55px\u201d \u0628\u0627\u0631\u0688\u0631_color_bottom=\u201d#29327a\u201d \u0644\u0627\u06a9=\u201d\u0622\u0641 global_colors_info=\u201d{}\u201d][et_pb_fullwidth_header _builder_version=\u201d4.16\u2033 title_font=\u201d||||||||\u201d 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header_font_size_last_edited=&quot;&quot;on_font_top_2tablet 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_tablet=heder_4_line 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>\u067e\u0631\u0648\u0679\u0648\u06a9\u0648\u0644 \u0628\u0631\u0627\u0626\u06d2 \u06a9\u0644\u0686\u0631\u0646\u06af \u0679\u0631\u0627\u0646\u0633\u0641\u0627\u0631\u0645\u0688 \u0644\u06cc\u0645\u0641\u0648\u0633\u0627\u0626\u0679\u0633<\/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\u201d|false custom_padding_tablet=&quot;|35px||35px||true&quot; custom_padding_phone=&quot;&quot; animation_direction=&quot;top&quot; global_colors_info=&quot;{}&quot;][et_pb_column_inner saved_specialty_column_type=&quot;3_4\u2033 _builder_version=&quot;6|4&quot;_builder_version=&quot;| global_colors_info=&quot;{}&quot; custom_padding__hover=&quot;|||&quot;][et_pb_text _builder_version=&quot;4.27.4\u2033 background_size=&quot;initial&quot; background_position=&quot;top_left&quot; background_repeat=&quot;repeat&quot; custom_padding=&quot;||0px\u2033||false=&quot;dfalse| global_colors_info=&quot;{}&quot; sticky_enabled=&quot;0\u2033]<\/p>\n<h4 style=\"text-align: center;\"><strong>\u0644\u06cc\u0645\u0641\u0648\u0628\u0644\u0627\u0633\u0679\u0633 \u06a9\u06cc \u062b\u0642\u0627\u0641\u062a \u06a9\u0627 \u067e\u0631\u0648\u0679\u0648\u06a9\u0648\u0644<\/strong><\/h4>\n<h4><strong>\u06af\u0631\u0648\u062a\u06be \u0645\u06cc\u0688\u06cc\u0645<\/strong><\/h4>\n<p>RPMI 1640 - ThermoFisher # 11875<br \/>15% \u0641\u06cc\u0679\u0644 \u0628\u0648\u0648\u0627\u0626\u0646 \u0633\u06cc\u0631\u0645 (FBS) - \u062a\u06be\u0631\u0645\u0648 \u0641\u0634\u0631 #10437-028<br \/>1% (1X) Penicillin-Streptomycin (\u0627\u062e\u062a\u06cc\u0627\u0631\u06cc) - ThermoFisher #15140-122<\/p>\n<h4><strong><\/strong><\/h4>\n<h4><strong>\u0639\u0645\u0648\u0645\u06cc \u0646\u0648\u0679\u0633<br \/><\/strong><\/h4>\n<ul>\n<li>\u06af\u0631\u0648\u062a\u06be \u0645\u06cc\u0688\u06cc\u0645 \u06a9\u0648 \u06c1\u0645\u06cc\u0634\u06c1 \u0627\u0646\u06a9\u06cc\u0648\u0628\u06cc\u0679\u0631 \u0645\u06cc\u06ba 37 \u00b0 C\u060c 5% CO \u067e\u0631 \u0645\u062a\u0648\u0627\u0632\u0646 \u06c1\u0648\u0646\u0627 \u0686\u0627\u06c1\u0626\u06d2<sub>2<\/sub> \u0627\u0633\u062a\u0639\u0645\u0627\u0644 \u0633\u06d2 \u067e\u06c1\u0644\u06d2 30 \u0645\u0646\u0679 \u06a9\u06d2 \u0644\u0626\u06d2<\/li>\n<li>\u06c1\u0645\u06cc\u0634\u06c1 \u0648\u06cc\u0646\u0679\u06cc\u0688 \u06a9\u06cc\u067e\u0633 \u06a9\u06d2 \u0633\u0627\u062a\u06be \u0641\u0644\u0627\u0633\u06a9\u0633 \u0627\u0633\u062a\u0639\u0645\u0627\u0644 \u06a9\u0631\u06cc\u06ba\u06d4<\/li>\n<li>\u0641\u0644\u0627\u0633\u06a9 \u06a9\u0648 \u06c1\u0645\u06cc\u0634\u06c1 \u0633\u06cc\u062f\u06be\u06cc \u067e\u0648\u0632\u06cc\u0634\u0646 \u0645\u06cc\u06ba \u0644\u06af\u0627\u0626\u06cc\u06ba\u06d4<\/li>\n<li>T25 \u0641\u0644\u0627\u0633\u06a9 \u0645\u06cc\u06ba 20 \u0645\u0644\u06cc \u0644\u06cc\u0679\u0631 \u0633\u06d2 \u0632\u06cc\u0627\u062f\u06c1 \u0627\u0633\u062a\u0639\u0645\u0627\u0644 \u0646\u06c1\u06cc\u06ba \u06a9\u0631\u0646\u0627 \u0686\u0627\u06c1\u0626\u06d2\u06d4<\/li>\n<\/ul>\n<h4><strong><\/strong><\/h4>\n<h4><strong>\u0645\u0646\u062c\u0645\u062f \u062e\u0644\u06cc\u0648\u06ba \u06a9\u0648 \u067e\u06af\u06be\u0644\u0627\u0646\u0627<br \/><\/strong><\/h4>\n<p>\u0645\u0646\u062c\u0645\u062f \u062e\u0644\u06cc\u0627\u062a \u062e\u0634\u06a9 \u0628\u0631\u0641 \u067e\u0631 1 \u0645\u0644\u06cc \u0644\u06cc\u0679\u0631 \u0627\u06cc\u0644\u06cc \u06a9\u0648\u0679\u0633 \u0645\u06cc\u06ba \u0648\u0635\u0648\u0644 \u06a9\u06cc\u06d2 \u062c\u0627\u0626\u06cc\u06ba \u06af\u06d2\u06d4 \u0633\u06cc\u0644 \u06a9\u0644\u0686\u0631 \u06a9\u06d2 \u0644\u06cc\u06d2 \u067e\u06af\u06be\u0644\u0646\u06d2 \u062a\u06a9 \u0633\u06cc\u0644\u0632 \u06a9\u0648 \u0645\u0646\u062c\u0645\u062f \u0631\u06a9\u06be\u06cc\u06ba \u06cc\u0627 \u0645\u0627\u0626\u0639 \u0646\u0627\u0626\u0679\u0631\u0648\u062c\u0646 \u0627\u0633\u0679\u0648\u0631\u06cc\u062c \u0645\u06cc\u06ba \u0631\u06a9\u06be\u06cc\u06ba \u0627\u06af\u0631 \u06a9\u0644\u0686\u0631\u0632 \u0627\u0628\u06be\u06cc \u0634\u0631\u0648\u0639 \u0646\u06c1 \u06c1\u0648\u06ba\u06d4 \u062a\u0627\u06c1\u0645\u060c \u06cc\u06c1 \u0633\u0641\u0627\u0631\u0634 \u06a9\u06cc \u062c\u0627\u062a\u06cc \u06c1\u06d2 \u06a9\u06c1 \u0627\u06af\u0631 \u0622\u067e \u0628\u0639\u062f \u0645\u06cc\u06ba \u0627\u0633\u062a\u0639\u0645\u0627\u0644 \u06a9\u06d2 \u0644\u06cc\u06d2 \u062e\u0644\u06cc\u0648\u06ba \u06a9\u0648 \u0630\u062e\u06cc\u0631\u06c1 \u06a9\u0631\u0646\u06d2 \u062c\u0627 \u0631\u06c1\u06d2 \u06c1\u06cc\u06ba\u060c \u062a\u0648 \u062a\u0627\u0632\u06c1 \u0630\u062e\u06cc\u0631\u06c1 \u0628\u0691\u06be\u0627\u0626\u06cc\u06ba \u0627\u0648\u0631 \u0645\u062a\u0639\u062f\u062f ampoules \u06a9\u0648 \u0645\u0646\u062c\u0645\u062f \u06a9\u0631\u06cc\u06ba\u06d4 \u062e\u0644\u06cc\u0627\u062a 45% RPMI-1640\u060c 50% FBS\u060c \u0627\u0648\u0631 5% DMSO (\u0679\u0634\u0648 \u06a9\u0644\u0686\u0631 \u06af\u0631\u06cc\u0688) \u0645\u06cc\u06ba \u06a9\u0631\u06cc\u0648\u067e\u0631\u06cc\u0632\u0631\u0688 \u06c1\u06cc\u06ba\u06d4<\/p>\n<ol>\n<li style=\"list-style-type: none;\">\n<ol>\n<li>RPMI-1640 \u06a9\u06d2 10 \u0645\u0644\u06cc \u0644\u06cc\u0679\u0631\u060c 15% \u0641\u06cc\u0679\u0644 \u0628\u0648\u0648\u0627\u0626\u0646 \u0633\u06cc\u0631\u0645 (FBS)\u060c \u00b1 \u0627\u06cc\u0646\u0679\u06cc \u0628\u0627\u0626\u06cc\u0648\u0679\u06a9\u0633 \u06a9\u0648 T-25 \u0641\u0644\u0627\u0633\u06a9 \u0645\u06cc\u06ba \u0648\u06cc\u0646\u0679\u06cc\u0688 \u06a9\u06cc\u067e \u06a9\u06d2 \u0633\u0627\u062a\u06be \u0631\u06a9\u06be\u06cc\u06ba\u06d4<\/li>\n<li>\u0645\u06cc\u0688\u06cc\u0627 \u06a9\u0648 5% CO \u06a9\u06d2 \u0633\u0627\u062a\u06be 37 \u00b0 C \u0646\u0645\u06cc \u0648\u0627\u0644\u06d2 \u0627\u0646\u06a9\u06cc\u0648\u0628\u06cc\u0679\u0631 \u0645\u06cc\u06ba \u0645\u062a\u0648\u0627\u0632\u0646 \u06c1\u0648\u0646\u06d2 \u062f\u06cc\u06ba\u06d4<sub>2<\/sub> 30 \u0645\u0646\u0679 \u06a9\u06d2 \u0644\u0626\u06d2 \u06c1\u0648\u0627 \u0645\u06cc\u06ba. \u0641\u0644\u0627\u0633\u06a9\u0633 \u06a9\u0648 \u0633\u06cc\u062f\u06be\u06cc \u067e\u0648\u0632\u06cc\u0634\u0646 \u0645\u06cc\u06ba \u0644\u06af\u0627\u0646\u0627 \u0686\u0627\u06c1\u06cc\u06d2\u06d4<\/li>\n<li>\u06c1\u0631 \u0627\u06cc\u06a9 \u0627\u06cc\u0645\u067e\u0648\u0644 \u06cc\u0627 \u0645\u0646\u062c\u0645\u062f \u062e\u0644\u06cc\u0648\u06ba \u06a9\u06cc \u0634\u06cc\u0634\u06cc \u06a9\u0648 37 \u00b0 C \u067e\u0627\u0646\u06cc \u06a9\u06d2 \u063a\u0633\u0644 \u0645\u06cc\u06ba \u06cc\u0627 \u0646\u06cc\u0645 \u06af\u0631\u0645 \u067e\u0627\u0646\u06cc \u06a9\u06d2 \u0627\u06cc\u06a9 \u0628\u06cc\u06a9\u0631 \u0645\u06cc\u06ba\u060c \u0627\u06cc\u06a9 \u0648\u0642\u062a \u0645\u06cc\u06ba \u0627\u06cc\u06a9 \u067e\u06af\u06be\u0644\u0627 \u062f\u06cc\u06ba\u06d4<\/li>\n<li>70% \u0627\u06cc\u062a\u06be\u0646\u0648\u0644 \u06a9\u06d2 \u0633\u0627\u062a\u06be \u0627\u06cc\u0645\u067e\u0648\u0644 \u06cc\u0627 \u0634\u06cc\u0634\u06cc \u06a9\u06d2 \u0628\u0627\u06c1\u0631 \u06a9\u0648 \u062c\u0644\u062f\u06cc \u0633\u06d2 \u0635\u0627\u0641 \u06a9\u0631\u06cc\u06ba\u06d4 \u0627\u06af\u0631 \u062e\u0644\u06cc\u06d2 \u0634\u06cc\u0634\u06d2 \u06a9\u06d2 \u0627\u06cc\u0645\u067e\u0648\u0644 \u0645\u06cc\u06ba \u06c1\u06cc\u06ba \u062a\u0648\u060c \u0627\u0646\u06af\u0644\u06cc\u0648\u06ba \u06a9\u0648 \u0679\u0648\u0679\u06d2 \u06c1\u0648\u0626\u06d2 \u0634\u06cc\u0634\u06d2 \u0633\u06d2 \u0628\u0686\u0627\u0646\u06d2 \u06a9\u06d2 \u0644\u06cc\u06d2 \u0627\u0633 \u0628\u0627\u062a \u06a9\u0648 \u06cc\u0642\u06cc\u0646\u06cc \u0628\u0646\u0627\u062a\u06d2 \u06c1\u0648\u0626\u06d2 \u0627\u06cc\u0645\u067e\u0648\u0644 \u06a9\u0648 \u0627\u062d\u062a\u06cc\u0627\u0637 \u0633\u06d2 \u062a\u0648\u0691 \u062f\u06cc\u06ba\u06d4<\/li>\n<li>1 \u0645\u0644\u06cc \u0644\u06cc\u0679\u0631 \u0645\u0646\u062c\u0645\u062f \u062e\u0644\u06cc\u0648\u06ba \u06a9\u0648 \u062c\u0631\u0627\u062b\u06cc\u0645 \u0633\u06d2 \u067e\u0627\u06a9 \u067e\u0627\u0626\u067e\u06cc\u0679 \u0633\u06d2 \u06c1\u0679\u0627\u0626\u06cc\u06ba \u0627\u0648\u0631 10 \u0645\u0644\u06cc \u0644\u06cc\u0679\u0631 \u0645\u06cc\u0688\u06cc\u0645 \u06a9\u06d2 \u0633\u0627\u062a\u06be T-25 \u0641\u0644\u0627\u0633\u06a9 \u0645\u06cc\u06ba \u0631\u06a9\u06be\u06cc\u06ba\u06d4 \u0648\u0627\u0636\u062d \u0637\u0648\u0631 \u067e\u0631 \u06c1\u0631 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style=\"list-style-type: none;\">\n<ol>\n<li>\u067e\u06af\u06be\u0644\u0646\u06d2 \u06a9\u06d2 \u062a\u06cc\u0646 \u0633\u06d2 \u0686\u0627\u0631 \u062f\u0646 \u0628\u0639\u062f\u060c \u062e\u0644\u06cc\u0627\u062a \u0634\u0645\u0627\u0631 \u06a9\u0631\u06cc\u06ba\u06d4 \u062e\u0644\u06cc\u06d2 \u063a\u06cc\u0631 \u0645\u0627\u0646\u0646\u06d2 \u0648\u0627\u0644\u06d2 \u06c1\u0648\u062a\u06d2 \u06c1\u06cc\u06ba \u0627\u0648\u0631 \u06af\u0686\u06be\u0648\u06ba \u0645\u06cc\u06ba \u0628\u0691\u06be\u062a\u06d2 \u06c1\u06cc\u06ba\u06d4 \u06c1\u0644\u06a9\u06cc \u067e\u0627\u0626\u067e\u0679\u0646\u06af \u06a9\u06d2 \u0630\u0631\u06cc\u0639\u06d2 \u06a9\u0644\u067e\u0633 \u06a9\u0648 \u062a\u0648\u0691 \u062f\u06cc\u06ba\u06d4<\/li>\n<li>\u062e\u0644\u06cc\u0648\u06ba \u06a9\u06cc \u06af\u0646\u062a\u06cc \u06a9\u0631\u06cc\u06ba\u06d4<\/li>\n<li>\u0645\u062b\u0627\u0644\u06cc \u0637\u0648\u0631 \u067e\u0631\u060c \u0633\u06cc\u0644 \u06a9\u06cc \u062d\u0631\u0627\u0633\u062a\u06cc 2 x 10 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\u062e\u0644\u06cc\u0627\u062a\/\u0645\u0644\u06cc\u06d4<\/li>\n<li>\u0633\u06cc\u0644\u0632 \u06a9\u0648 \u062a\u0642\u0631\u06cc\u0628\u0627\u064b 5 x 10 \u067e\u0631 \u0645\u0646\u062c\u0645\u062f \u06a9\u06cc\u0627 \u062c\u0627\u0646\u0627 \u0686\u0627\u06c1\u06cc\u06d2\u06d4<sup>6<\/sup> \u062e\u0644\u06cc\u0627\u062a\/\u0645\u0644\u06cc\u06d4 \u0633\u06cc\u0644 \u0641\u0631\u06cc\u0632\u0646\u06af \u067e\u0631\u0648\u0679\u0648\u06a9\u0648\u0644 \u067e\u0631 \u0639\u0645\u0644 \u06a9\u0631\u06cc\u06ba\u06d4<\/li>\n<\/ol>\n<\/li>\n<\/ol>\n<h4><strong><\/strong><\/h4>\n<h4><strong>\u0645\u0646\u062c\u0645\u062f \u0644\u06cc\u0645\u0641\u0648\u0628\u0644\u0627\u0633\u0679\u0633<\/strong><\/h4>\n<ol>\n<li style=\"list-style-type: none;\">\n<ol>\n<li>\u0633\u06cc\u0644\u0632 \u06a9\u0648 \u062a\u0642\u0631\u06cc\u0628\u0627\u064b 5 x 10 \u067e\u0631 \u0645\u0646\u062c\u0645\u062f \u06a9\u06cc\u0627 \u062c\u0627\u0646\u0627 \u0686\u0627\u06c1\u06cc\u06d2\u06d4<sup>6<\/sup> 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saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.27.4\u2033 _module_preset=\u201ddefault\u201d global_colors_info=\u201d{}\u201d][et_pton button_url=\u201dhttps:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2025\/02\/Lymphoblast-protocol-1-22-25.pdf\u201d button_text=\u201dProtocol for Culturing Lymphoblasts\u201d admin_label=\u201dProtocol for Culturing Lymphoblasts.2\u201d_Lymphoblasts. _module_preset=\u201ddefault\u201d button_text_color=\u201d#FFFFFF\u201d \u0628\u0679\u0646_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||0|| custom_padding_tablet=\u201d|35px||35px||true\u201d custom_padding_phone=\u201d\u201dborder_width_top=\u201d10px\u201d \u0628\u0627\u0631\u0688\u0631_color_top=\u201d#8fd2ed\u201d global_colors_info=\u201d{}\u201d][et_pb_column_inner saved_specialty\u2033_specialty=3&quot;_specialty&quot; _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=&quot;{}&quot; custom_padding__hover=&quot;|||&quot;][et_pb_text _builder_version=&quot;4.16\u2033 global_colors_info=&quot;{}&quot;]<\/p>\n<h4><strong>\u0645\u0632\u06cc\u062f \u0645\u0639\u0644\u0648\u0645\u0627\u062a \u06a9\u06d2 \u0644\u06cc\u06d2 \u0628\u0631\u0627\u0626\u06d2 \u0645\u06c1\u0631\u0628\u0627\u0646\u06cc \u0631\u0627\u0628\u0637\u06c1 \u06a9\u0631\u06cc\u06ba:<\/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_info=\u201d1 inner type=\u201d1 saved_specialty_column_type=\u201d3_4\u2033 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|20px||\u201d custom_padding_tablet=&quot;||20px&quot; custom_padding_phone=&quot;&quot; custom_padding_last_edited=&quot;on|desktop&quot; global_colors_info=&quot;{}&quot; custom_padding__hover=&quot;|||&quot;][et_pb_text _builder_version=&quot;4.19.5\u2033 _color&quot;=&quot;_color&quot; \u0645\u06cc\u06ba<\/p>\n<h6>\u0644\u06cc\u0633\u0644\u06cc \u0628\u06cc \u06af\u0648\u0631\u0688\u0646\u060c \u0627\u06cc\u0645 \u0688\u06cc\u060c \u067e\u06cc \u0627\u06cc\u0686 \u0688\u06cc<\/h6>\n<p>\u0628\u0631\u0627\u0624\u0646 \u06cc\u0648\u0646\u06cc\u0648\u0631\u0633\u0679\u06cc \u06a9\u06d2 \u067e\u06cc\u0688\u06cc\u0627\u0679\u0631\u06a9\u0633 \u0631\u06cc\u0633\u0631\u0686 \u0648\u0627\u0631\u0646 \u0627\u0644\u067e\u0631\u0679 \u0645\u06cc\u0688\u06cc\u06a9\u0644 \u0633\u06a9\u0648\u0644 \u06a9\u06d2 \u067e\u0631\u0648\u0641\u06cc\u0633\u0631 \u0627\u0648\u0631 \u0634\u0639\u0628\u06c1 \u0627\u0637\u0641\u0627\u0644\u060c \u06c1\u0627\u0633\u0628\u0631\u0648 \u0686\u0644\u0688\u0631\u0646 \u06c1\u0633\u067e\u062a\u0627\u0644\u060c \u067e\u0631\u0648\u0648\u06cc\u0688\u0646\u0633\u060c RI \u0688\u06cc\u067e\u0627\u0631\u0679\u0645\u0646\u0679 \u0622\u0641 \u0627\u06cc\u0646\u0633\u062a\u06be\u06cc\u0632\u06cc\u0627\u060c \u0686\u0644\u0688\u0631\u0646 \u06c1\u0633\u067e\u062a\u0627\u0644 \u0628\u0648\u0633\u0679\u0646 \u0627\u0648\u0631 \u06c1\u0627\u0631\u0648\u0631\u0688 \u0645\u06cc\u0688\u06cc\u06a9\u0644 \u0633\u06a9\u0648\u0644\u060c \u0628\u0648\u0633\u0679\u0646\u060c \u0627\u06cc\u0645 \u0627\u06d2 \u0645\u06cc\u0688\u06cc\u06a9\u0644 \u0688\u0627\u0626\u0631\u06cc\u06a9\u0679\u0631\u060c \u062f\u06cc \u067e\u0631\u0648\u062c\u06cc\u0631\u06cc\u0627 \u0631\u06cc\u0633\u0631\u0686 \u0641\u0627\u0624\u0646\u0688\u06cc\u0634\u0646\u06d4<\/p>\n<p>\u0641\u0648\u0646: 978-535-2594<br \/>\u0641\u06cc\u06a9\u0633: 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=\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>\u0648\u06cc\u0646\u0688\u06cc \u0646\u0648\u0631\u0633<\/span><\/h6>\n<div><span>\u067e\u06cc \u0622\u0631 \u0627\u06cc\u0641 \u0633\u06cc\u0644 \u0627\u0648\u0631 \u0679\u0634\u0648 \u0628\u06cc\u0646\u06a9<br 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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":"<!-- 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