{"id":1006,"date":"2017-02-27T21:20:56","date_gmt":"2017-02-27T21:20:56","guid":{"rendered":"https:\/\/www.progeriaresearch.org\/?page_id=1006"},"modified":"2024-10-23T11:05:53","modified_gmt":"2024-10-23T15:05:53","slug":"fibroblast-cell-culture-protocols","status":"publish","type":"page","link":"https:\/\/www.progeriaresearch.org\/kn\/fibroblast-cell-culture-protocols\/","title":{"rendered":"\u0cab\u0cc8\u0cac\u0ccd\u0cb0\u0cca\u0cac\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c9f\u0ccd \u0cb8\u0cc6\u0cb2\u0ccd \u0c95\u0cb2\u0ccd\u0c9a\u0cb0\u0ccd \u0caa\u0ccd\u0cb0\u0ccb\u0c9f\u0ccb\u0c95\u0cbe\u0cb2\u0ccd\u200c\u0c97\u0cb3\u0cc1"},"content":{"rendered":"

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\u0cab\u0cc8\u0cac\u0ccd\u0cb0\u0cca\u0cac\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c9f\u0ccd \u0c95\u0ccb\u0cb6<\/h1>\n

\u0cb8\u0c82\u0cb8\u0ccd\u0c95\u0cc3\u0ca4\u0cbf \u0caa\u0ccd\u0cb0\u0ccb\u0c9f\u0ccb\u0c95\u0cbe\u0cb2\u0ccd\u0c97\u0cb3\u0cc1<\/h1>\n

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\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_\u201d51slide=\u201dT2P3 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_4type=\u201d3 _builder_version=\u201d4.16\u2033 custom_padding=\u201d|||\u201d global_colors_info=\u201d{}\u201d custom_padding__hover=\u201d|||\u201d][et_pb_text admin_label=\u201dProgeria \u0cb0\u0cbf\u0cb8\u0cb0\u0ccd\u0c9a\u0ccd \u0cab\u0ccc\u0c82\u0ca1\u0cc7\u0cb6\u0ca8\u0ccd\u200c\u0ca8 \u0c95\u0cc6\u0cb2\u0cb8\u2026\u201d _builder_version=\u201d4.16\u2033 header_2_line_height=\u201d1.2em\u201d background_layout=\u201dx= custom_5 ="||" 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| 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_stonedsitize\u201dphone 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

\u0c89\u0caa\u0cb8\u0c82\u0cb8\u0ccd\u0c95\u0cc3\u0ca4\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0c98\u0ca8\u0cc0\u0c95\u0cb0\u0cbf\u0cb8\u0cc1\u0cb5 \u0cab\u0cc8\u0cac\u0ccd\u0cb0\u0cca\u0cac\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c9f\u0ccd\u200c\u0c97\u0cb3\u0cbf\u0c97\u0cbe\u0c97\u0cbf \u0caa\u0ccd\u0cb0\u0ccb\u0c9f\u0ccb\u0c95\u0cbe\u0cb2\u0ccd<\/strong><\/h2>\n

[\/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|20px|35px||true\u201d custom_padding_phone=\u201d|35px|11px|35px|false|true\u201d animation_direction=\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=\u201d4.16=_ \u0cb9\u0cbf\u0ca8\u0ccd\u0ca8\u0cc6\u0cb2\u0cc6\u201d \u0cb9\u0cbf\u0ca8\u0ccd\u0ca8\u0cc6\u0cb2\u0cc6 background_repeat=\u201drepeat\u201d custom_padding=\u201d||24px||\u201d global_colors_info=\u201d{}\u201d]<\/p>\n

\u0cac\u0cc6\u0cb3\u0cb5\u0ca3\u0cbf\u0c97\u0cc6 \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae<\/h4>\n

DMEM - \u0ca5\u0cb0\u0ccd\u0cae\u0ccb\u0cab\u0cbf\u0cb6\u0cb0\u0ccd #11960-044 (\u0c8e\u0cb2\u0ccd-\u0c97\u0ccd\u0cb2\u0cc1\u0c9f\u0cbe\u0cae\u0cbf\u0ca8\u0ccd \u0c87\u0cb2\u0ccd\u0cb2\u0ca6\u0cc6 \u0cb9\u0cc6\u0c9a\u0ccd\u0c9a\u0cbf\u0ca8 \u0c97\u0ccd\u0cb2\u0cc2\u0c95\u0ccb\u0cb8\u0ccd)<\/p>\n

15% FBS \u0cab\u0cc6\u0c9f\u0cb2\u0ccd \u0cac\u0ccb\u0cb5\u0cbf\u0ca8\u0ccd \u0cb8\u0cc0\u0cb0\u0cae\u0ccd - \u0ca5\u0cb0\u0ccd\u0cae\u0ccb\u0cab\u0cbf\u0cb6\u0cb0\u0ccd #10437-028<\/p>\n

1% (1X) \u0caa\u0cc6\u0ca8\u0ccd\u0cb8\u0cbf\u0cb2\u0cbf\u0ca8\u0ccd-\u0cb8\u0ccd\u0c9f\u0ccd\u0cb0\u0cc6\u0caa\u0ccd\u0c9f\u0cca\u0cae\u0cc8\u0cb8\u0cbf\u0ca8\u0ccd - \u0ca5\u0cb0\u0ccd\u0cae\u0ccb\u0cab\u0cbf\u0cb6\u0cb0\u0ccd #15140-122<\/p>\n

1% (1X) GlutaMAX \u2013 ThermoFisher #35050-061<\/p>\n

[\/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||=20pdpx\u201d \u0c95\u0cb8\u0ccd\u0c9f\u0cae\u0ccd 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=\u201d4.16\u2033background_size=\u201dinitial\u201d background_position=\u201dtop_left\u201dback_peleat custom_padding=\u201d||24px||\u201d global_colors_info=\u201d{}\u201d]<\/p>\n

\u0c9f\u0ccd\u0cb0\u0cbf\u0caa\u0ccd\u0cb8\u0cbf\u0ca8\u0ccd<\/strong><\/h4>\n

\u0c9f\u0ccd\u0cb0\u0cbf\u0caa\u0ccd\u0cb8\u0cbf\u0ca8\u0ccd EDTA C 0.25% (ThermoFisher #25200-056)<\/p>\n

\u0cb9\u0ccd\u0caf\u0cbe\u0c82\u0c95\u0ccd\u0cb8\u0ccd \u0cb8\u0cae\u0ca4\u0ccb\u0cb2\u0cbf\u0ca4 \u0c89\u0caa\u0ccd\u0caa\u0cc1 \u0caa\u0cb0\u0cbf\u0cb9\u0cbe\u0cb0<\/strong><\/h4>\n

HBSS- (ThermoFisher #14170 (1X) (-) \u0c95\u0ccd\u0caf\u0cbe\u0cb2\u0ccd\u0cb8\u0cbf\u0caf\u0c82 \u0c95\u0ccd\u0cb2\u0ccb\u0cb0\u0cc8\u0ca1\u0ccd, (-) \u0cae\u0cc6\u0c97\u0ccd\u0ca8\u0cc0\u0cb8\u0cbf\u0caf\u0cae\u0ccd \u0c95\u0ccd\u0cb2\u0ccb\u0cb0\u0cc8\u0ca1\u0ccd, (-) \u0cae\u0cc6\u0c97\u0ccd\u0ca8\u0cc0\u0cb8\u0cbf\u0caf\u0cae\u0ccd \u0cb8\u0cb2\u0ccd\u0cab\u0cc7\u0c9f\u0ccd<\/p>\n

\u0c98\u0ca8\u0cc0\u0c95\u0cb0\u0ca3\u0c95\u0ccd\u0c95\u0cbe\u0c97\u0cbf DMSO<\/strong><\/h4>\n

<\/u><\/strong>\u0cb8\u0cc6\u0cb2\u0ccd \u0c95\u0cb2\u0ccd\u0c9a\u0cb0\u0ccd \u0c97\u0ccd\u0cb0\u0cc7\u0ca1\u0ccd DMSO - \u0cb8\u0cbf\u0c97\u0ccd\u0cae\u0cbe #D2438<\/p>\n

[\/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_margin=\u201d||35px\u201d custom_padding=\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_\u201d_3 \u0cb5\u0cbf\u0cb6\u0cc7\u0cb7\u0ca4\u0cc64 _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

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PRF \u0cb8\u0cc6\u0cb2\u0ccd \u0cb2\u0cc8\u0ca8\u0ccd\u200c\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0c95\u0cb0\u0c97\u0cbf\u0cb8\u0cc1\u0cb5\u0cc1\u0ca6\u0cc1<\/strong><\/h4>\n
    \n
  1. 37\u02daC \u0ca8\u0cc0\u0cb0\u0cbf\u0ca8 \u0cb8\u0ccd\u0ca8\u0cbe\u0ca8\u0ca6\u0cb2\u0ccd\u0cb2\u0cbf \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb5\u0cc7\u0c97\u0cb5\u0cbe\u0c97\u0cbf \u0c95\u0cb0\u0c97\u0cbf\u0cb8\u0cbf.<\/li>\n
  2. 70% \u0c8e\u0ca5\u0cc6\u0ca8\u0cbe\u0cb2\u0ccd\u0ca8\u0cca\u0c82\u0ca6\u0cbf\u0c97\u0cc6 \u0cac\u0cbe\u0c9f\u0cb2\u0cbf\u0caf \u0cb9\u0cca\u0cb0\u0cad\u0cbe\u0c97\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0c92\u0cb0\u0cc6\u0cb8\u0cbf.<\/li>\n
  3. \u0c95\u0cb0\u0c97\u0cbf\u0ca6 \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 5 \u0cae\u0cbf\u0cb2\u0cbf \u0cac\u0cc6\u0cb3\u0cb5\u0ca3\u0cbf\u0c97\u0cc6 \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb9\u0cca\u0c82\u0ca6\u0cbf\u0cb0\u0cc1\u0cb5 T25 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd\u200c\u0c97\u0cc6 \u0cb5\u0cb0\u0ccd\u0c97\u0cbe\u0caf\u0cbf\u0cb8\u0cbf.<\/li>\n
  4. \u0cb0\u0cbe\u0ca4\u0ccd\u0cb0\u0cbf\u0caf\u0cbf\u0ca1\u0cc0 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd \u0c85\u0ca8\u0ccd\u0ca8\u0cc1 37\u02daC \u0c87\u0ca8\u0ccd\u0c95\u0ccd\u0caf\u0cc1\u0cac\u0cc7\u0c9f\u0cb0\u0ccd\u0ca8\u0cb2\u0ccd\u0cb2\u0cbf \u0c87\u0cb0\u0cbf\u0cb8\u0cbf.<\/li>\n
  5. \u0cae\u0cb0\u0cc1\u0ca6\u0cbf\u0ca8, \u0c9c\u0cc0\u0cb5\u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cc1 \u0cb2\u0c97\u0ca4\u0ccd\u0ca4\u0cbf\u0cb8\u0cbf\u0cb0\u0cc1\u0cb5\u0cc1\u0ca6\u0ca8\u0ccd\u0ca8\u0cc1 \u0c96\u0c9a\u0cbf\u0ca4\u0caa\u0ca1\u0cbf\u0cb8\u0cbf\u0c95\u0cca\u0cb3\u0ccd\u0cb3\u0cb2\u0cc1 \u0cb8\u0cc2\u0c95\u0ccd\u0cb7\u0ccd\u0cae\u0ca6\u0cb0\u0ccd\u0cb6\u0c95\u0ca6 \u0c85\u0ca1\u0cbf\u0caf\u0cb2\u0ccd\u0cb2\u0cbf \u0c97\u0cae\u0ca8\u0cbf\u0cb8\u0cbf.<\/li>\n
  6. \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0ca4\u0cc6\u0c97\u0cc6\u0ca6\u0cc1\u0cb9\u0cbe\u0c95\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0ca4\u0cbe\u0c9c\u0cbe \u0cac\u0cc6\u0cb3\u0cb5\u0ca3\u0cbf\u0c97\u0cc6 \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0ca6\u0cca\u0c82\u0ca6\u0cbf\u0c97\u0cc6 \u0cac\u0ca6\u0cb2\u0cbe\u0caf\u0cbf\u0cb8\u0cbf.<\/li>\n<\/ol>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n
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    \u0c89\u0caa\u0cb8\u0c82\u0cb8\u0ccd\u0c95\u0cc3\u0ca4\u0cbf PRF \u0cb8\u0cc6\u0cb2\u0ccd \u0cb2\u0cc8\u0ca8\u0ccd\u200c\u0c97\u0cb3\u0cc1<\/strong><\/h4>\n
      \n
    1. \u0cb8\u0c82\u0c97\u0cae\u0cb5\u0cbe\u0ca6\u0cbe\u0c97 \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb5\u0cbf\u0cad\u0c9c\u0cbf\u0cb8\u0cac\u0cc7\u0c95\u0cc1.<\/li>\n
    2. \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb5\u0cbf\u0cad\u0c9c\u0cbf\u0cb8\u0cb2\u0cc1 (\u0caa\u0cb0\u0cbf\u0cae\u0cbe\u0ca3\u0c97\u0cb3\u0cc1 \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cc1 T25 \u0ca8\u0cb2\u0ccd\u0cb2\u0cbf\u0cb5\u0cc6 \u0c8e\u0c82\u0ca6\u0cc1 \u0c8a\u0cb9\u0cbf\u0cb8\u0cb2\u0cbe\u0c97\u0cbf\u0ca6\u0cc6):
      a) \u0cac\u0cb0\u0ca1\u0cbe\u0ca6 \u0ca4\u0c82\u0ca4\u0ccd\u0cb0\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0cac\u0cb3\u0cb8\u0cbf, \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0ca4\u0cc6\u0c97\u0cc6\u0ca6\u0cc1\u0cb9\u0cbe\u0c95\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 2-3 \u0cae\u0cbf\u0cb2\u0cbf \u0cb8\u0ccd\u0c9f\u0cc6\u0cb0\u0cc8\u0cb2\u0ccd HBSS \u0ca8\u0cca\u0c82\u0ca6\u0cbf\u0c97\u0cc6 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd \u0c85\u0ca8\u0ccd\u0ca8\u0cc1 \u0ca4\u0cca\u0cb3\u0cc6\u0caf\u0cbf\u0cb0\u0cbf. HBSS \u0ca4\u0cc6\u0c97\u0cc6\u0ca6\u0cc1\u0cb9\u0cbe\u0c95\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0ca4\u0ccd\u0caf\u0c9c\u0cbf\u0cb8\u0cbf.
      \u0cac\u0cbf) 1 \u0cae\u0cbf\u0cb2\u0cbf \u0c9f\u0ccd\u0cb0\u0cbf\u0caa\u0ccd\u0cb8\u0cbf\u0ca8\u0ccd \u0cb8\u0cc7\u0cb0\u0cbf\u0cb8\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 2-3 \u0ca8\u0cbf\u0cae\u0cbf\u0cb7\u0c97\u0cb3 \u0c95\u0cbe\u0cb2 \u0c95\u0cbe\u0cb5\u0cc1\u0c95\u0cca\u0ca1\u0cbf. \u0c9c\u0cc0\u0cb5\u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cc1 \u0cb8\u0cc1\u0ca4\u0ccd\u0ca4\u0cc1\u0cb5\u0cb0\u0cbf\u0caf\u0cb2\u0cc1, \u0c8e\u0ca4\u0ccd\u0ca4\u0cb2\u0cc1 \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0ca4\u0cc7\u0cb2\u0cb2\u0cc1 \u0caa\u0ccd\u0cb0\u0cbe\u0cb0\u0c82\u0cad\u0cbf\u0cb8\u0cbf\u0cb5\u0cc6\u0caf\u0cc7 \u0c8e\u0c82\u0ca6\u0cc1 \u0ca8\u0cbf\u0cb0\u0ccd\u0ca7\u0cb0\u0cbf\u0cb8\u0cb2\u0cc1 \u0ca4\u0cb2\u0cc6\u0c95\u0cc6\u0cb3\u0c97\u0cbe\u0ca6 \u0cb8\u0cc2\u0c95\u0ccd\u0cb7\u0ccd\u0cae\u0ca6\u0cb0\u0ccd\u0cb6\u0c95\u0ca6 \u0c85\u0ca1\u0cbf\u0caf\u0cb2\u0ccd\u0cb2\u0cbf \u0caa\u0cb0\u0cc0\u0c95\u0ccd\u0cb7\u0cbf\u0cb8\u0cac\u0cc7\u0c95\u0cc1. 3 \u0ca8\u0cbf\u0cae\u0cbf\u0cb7\u0c97\u0cb3 \u0ca8\u0c82\u0ca4\u0cb0 \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cac\u0cc7\u0cb0\u0ccd\u0caa\u0ca1\u0cbf\u0cb8\u0ca6\u0cbf\u0ca6\u0ccd\u0ca6\u0cb0\u0cc6, \u0ca8\u0cc0\u0cb5\u0cc1 \u0c87\u0ca8\u0ccd\u0ca8\u0cca\u0c82\u0ca6\u0cc1 1-2 \u0ca8\u0cbf\u0cae\u0cbf\u0cb7\u0c97\u0cb3 \u0c95\u0cbe\u0cb2 \u0c95\u0cbe\u0cb5\u0cc1\u0c95\u0cca\u0ca1\u0cac\u0cb9\u0cc1\u0ca6\u0cc1.
      \u0cb8\u0cbf) \u0c8e\u0cb2\u0ccd\u0cb2\u0cbe \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb9\u0cca\u0cb0\u0cb9\u0cbe\u0c95\u0cb2\u0cc1 \u0c95\u0ccd\u0caf\u0cbe\u0caa\u0ccd \u0c85\u0ca8\u0ccd\u0ca8\u0cc1 \u0cac\u0cbf\u0c97\u0cbf\u0c97\u0cca\u0cb3\u0cbf\u0cb8\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd \u0c85\u0ca8\u0ccd\u0ca8\u0cc1 \u0c85\u0c95\u0ccd\u0c95\u0caa\u0c95\u0ccd\u0c95\u0c95\u0ccd\u0c95\u0cc6 \u0ca8\u0cbf\u0ca7\u0cbe\u0ca8\u0cb5\u0cbe\u0c97\u0cbf \u0ca4\u0cbf\u0cb0\u0cc1\u0c97\u0cbf\u0cb8\u0cbf. \u0c85\u0c97\u0ca4\u0ccd\u0caf\u0cb5\u0cbf\u0ca6\u0ccd\u0ca6\u0cb0\u0cc6 \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb8\u0ca1\u0cbf\u0cb2\u0c97\u0cca\u0cb3\u0cbf\u0cb8\u0cb2\u0cc1 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd \u0c85\u0ca8\u0ccd\u0ca8\u0cc1 \u0cac\u0ca6\u0cbf\u0caf\u0cb2\u0ccd\u0cb2\u0cbf \u0cb2\u0c98\u0cc1\u0cb5\u0cbe\u0c97\u0cbf \u0c9f\u0ccd\u0caf\u0cbe\u0caa\u0ccd \u0cae\u0cbe\u0ca1\u0cac\u0cb9\u0cc1\u0ca6\u0cc1.
      \u0ca1\u0cbf) \u0c9f\u0ccd\u0cb0\u0cbf\u0caa\u0ccd\u0cb8\u0cbf\u0ca8\u0ccd \u0c85\u0ca8\u0ccd\u0ca8\u0cc1 \u0ca8\u0cbf\u0cb7\u0ccd\u0c95\u0ccd\u0cb0\u0cbf\u0caf\u0c97\u0cca\u0cb3\u0cbf\u0cb8\u0cb2\u0cc1 \u0c9c\u0cc0\u0cb5\u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cc1 \u0ca4\u0cc7\u0cb2\u0cc1\u0ca4\u0ccd\u0ca4\u0cbf\u0cb0\u0cc1\u0cb5 \u0ca4\u0c95\u0ccd\u0cb7\u0ca3 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd\u200c\u0c97\u0cc6 4 \u0cae\u0cbf\u0cb2\u0cbf \u0cb9\u0cca\u0cb8 \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb8\u0cc7\u0cb0\u0cbf\u0cb8\u0cbf. \u0c8e\u0cb2\u0ccd\u0cb2\u0cbe \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0caa\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c9f\u0cbf\u0c95\u0ccd\u200c\u0ca8\u0cbf\u0c82\u0ca6 \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0ca6\u0ccd\u0cb0\u0cbe\u0cb5\u0ca3\u0ca6\u0cb2\u0ccd\u0cb2\u0cbf \u0ca4\u0cca\u0cb3\u0cc6\u0caf\u0cb2\u0cc1 \u0cb9\u0cca\u0cb8 \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0ca6\u0cca\u0c82\u0ca6\u0cbf\u0c97\u0cc6 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd\u200c\u0ca8 \u0cac\u0ca6\u0cbf\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb9\u0cb2\u0cb5\u0cbe\u0cb0\u0cc1 \u0cac\u0cbe\u0cb0\u0cbf \u0ca4\u0cca\u0cb3\u0cc6\u0caf\u0cbf\u0cb0\u0cbf. \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb9\u0cca\u0c82\u0ca6\u0cbf\u0cb0\u0cc1\u0cb5 \u0c8e\u0cb2\u0ccd\u0cb2\u0cbe \u0ca6\u0ccd\u0cb0\u0cb5\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0ca4\u0cc6\u0c97\u0cc6\u0ca6\u0cc1\u0cb9\u0cbe\u0c95\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 15ml \u0cb6\u0c82\u0c95\u0cc1\u0cb5\u0cbf\u0ca8\u0cbe\u0c95\u0cbe\u0cb0\u0ca6 (\u0c85\u0ca5\u0cb5\u0cbe \u0ca8\u0cc0\u0cb5\u0cc1 3 \u0c85\u0ca5\u0cb5\u0cbe \u0cb9\u0cc6\u0c9a\u0ccd\u0c9a\u0cbf\u0ca8 \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd\u200c\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0caa\u0cc2\u0cb2\u0ccd \u0cae\u0cbe\u0ca1\u0cc1\u0ca4\u0ccd\u0ca4\u0cbf\u0ca6\u0ccd\u0ca6\u0cb0\u0cc6 50 \u0cae\u0cbf\u0cb2\u0cbf) \u0c97\u0cc6 \u0cb5\u0cb0\u0ccd\u0c97\u0cbe\u0caf\u0cbf\u0cb8\u0cbf.
      \u0c87) \u0c95\u0ccd\u0cb0\u0cbf\u0cae\u0cbf\u0ca8\u0cbe\u0cb6\u0c95 \u0ca4\u0c82\u0ca4\u0ccd\u0cb0\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0cac\u0cb3\u0cb8\u0cbf, \u0cb9\u0cc6\u0cae\u0ccb\u0cb8\u0cc8\u0c9f\u0ccb\u0cae\u0cc0\u0c9f\u0cb0\u0ccd\u200c\u0ca8\u0cb2\u0ccd\u0cb2\u0cbf \u0c8e\u0ca3\u0cbf\u0cb8\u0cb2\u0cc1 \u0c85\u0cb2\u0cbf\u0c95\u0ccb\u0c9f\u0ccd \u0c85\u0ca8\u0ccd\u0ca8\u0cc1 \u0ca4\u0cc6\u0c97\u0cc6\u0ca6\u0cc1\u0cb9\u0cbe\u0c95\u0cbf.
      \u0c8e\u0cab\u0ccd) \u0ca8\u0cc0\u0cb5\u0cc1 \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 \u0c8e\u0ca3\u0cbf\u0cb8\u0cc1\u0cb5\u0cbe\u0c97, \u0c89\u0cb3\u0cbf\u0ca6 \u0ca6\u0ccd\u0cb0\u0cbe\u0cb5\u0ca3\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0c95\u0ccd\u0cb2\u0cbf\u0ca8\u0cbf\u0c95\u0cb2\u0ccd \u0cb8\u0cc6\u0c82\u0c9f\u0ccd\u0cb0\u0cbf\u0cab\u0ccd\u0caf\u0cc2\u0c9c\u0ccd\u200c\u0ca8\u0cb2\u0ccd\u0cb2\u0cbf 5 \u0ca8\u0cbf\u0cae\u0cbf\u0cb7\u0c97\u0cb3 \u0c95\u0cbe\u0cb2 1000 \u0c86\u0cb0\u0ccd\u200c\u0caa\u0cbf\u0c8e\u0cae\u0ccd\u200c\u0ca8\u0cb2\u0ccd\u0cb2\u0cbf \u0ca4\u0cbf\u0cb0\u0cc1\u0c97\u0cbf\u0cb8\u0cac\u0cc7\u0c95\u0cc1.<\/li>\n
    3. \u0ca8\u0cbf\u0cae\u0ccd\u0cae \u0c95\u0ccb\u0cb6\u0c97\u0cb3 \u0cb8\u0c82\u0c96\u0ccd\u0caf\u0cc6\u0caf\u0ca8\u0ccd\u0ca8\u0cc1 \u0cb2\u0cc6\u0c95\u0ccd\u0c95\u0cbe\u0c9a\u0cbe\u0cb0 \u0cae\u0cbe\u0ca1\u0cbf\u0ca6 \u0ca8\u0c82\u0ca4\u0cb0, 2.5 x 10 \u0ca8\u0cb2\u0ccd\u0cb2\u0cbf \u0caa\u0ccd\u0cb2\u0cc7\u0c9f\u0ccd \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cc15<\/sup>\u00a0<\/span>T25 \u0cab\u0cbf\u0cb2\u0ccd\u0c9f\u0cb0\u0ccd \u0c9f\u0cbe\u0caa\u0ccd \u0cab\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c95\u0ccd \u0caa\u0ccd\u0cb0\u0ca4\u0cbf \u0c9c\u0cc0\u0cb5\u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cc1.<\/li>\n
    4. \u0ca4\u0cbe\u0c9c\u0cbe \u0cac\u0cc6\u0cb3\u0cb5\u0ca3\u0cbf\u0c97\u0cc6\u0caf \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0ca6\u0cca\u0c82\u0ca6\u0cbf\u0c97\u0cc6 \u0caa\u0ccd\u0cb0\u0ca4\u0cbf 2-3 \u0ca6\u0cbf\u0ca8\u0c97\u0cb3\u0cbf\u0c97\u0cca\u0cae\u0ccd\u0cae\u0cc6 \u0c9c\u0cc0\u0cb5\u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cbf\u0c97\u0cc6 \u0c86\u0cb9\u0cbe\u0cb0\u0cb5\u0ca8\u0ccd\u0ca8\u0cc1 \u0ca8\u0cc0\u0ca1\u0cac\u0cc7\u0c95\u0cc1.<\/li>\n<\/ol>\n

      \u0c98\u0ca8\u0cc0\u0c95\u0cb0\u0cbf\u0cb8\u0cc1\u0cb5\u0cbf\u0c95\u0cc6:
      <\/u><\/strong><\/h4>\n

      \u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0ca8\u0ccd\u0ca8\u0cc1 5 x 10 \u0c95\u0ccd\u0c95\u0cbf\u0c82\u0ca4 \u0c95\u0ca1\u0cbf\u0cae\u0cc6\u0caf\u0cbf\u0cb2\u0ccd\u0cb2\u0ca6\u0c82\u0ca4\u0cc6 \u0cab\u0ccd\u0cb0\u0cc0\u0c9c\u0ccd \u0cae\u0cbe\u0ca1\u0cac\u0cc7\u0c95\u0cc15<\/sup>\u00a0<\/span>10% DMSO \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 30% FBS \u0cb9\u0cca\u0c82\u0ca6\u0cbf\u0cb0\u0cc1\u0cb5 \u0cac\u0cc6\u0cb3\u0cb5\u0ca3\u0cbf\u0c97\u0cc6\u0caf \u0cae\u0cbe\u0ca7\u0ccd\u0caf\u0cae\u0ca6\u0cb2\u0ccd\u0cb2\u0cbf \u0c9c\u0cc0\u0cb5\u0c95\u0ccb\u0cb6\u0c97\u0cb3\u0cc1\/ml\/\u0c95\u0ccd\u0cb0\u0caf\u0ccb\u0cb5\u0cbf\u0caf\u0cb2\u0ccd \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0ca8\u0c82\u0ca4\u0cb0 \u0cb0\u0cbe\u0ca4\u0ccd\u0cb0\u0cbf -80\u02daC \u0ca8\u0cb2\u0ccd\u0cb2\u0cbf \u0c90\u0cb8\u0cca\u0caa\u0ccd\u0cb0\u0cca\u0caa\u0ca8\u0cbe\u0cb2\u0ccd \u0c98\u0ca8\u0cc0\u0c95\u0cb0\u0cbf\u0cb8\u0cc1\u0cb5 \u0c95\u0cca\u0ca0\u0ca1\u0cbf\u0caf\u0cb2\u0ccd\u0cb2\u0cbf \u0c87\u0cb0\u0cbf\u0cb8\u0cb2\u0cbe\u0c97\u0cc1\u0ca4\u0ccd\u0ca4\u0ca6\u0cc6. \u0cae\u0cb0\u0cc1\u0ca6\u0cbf\u0ca8 \u0ca6\u0ccd\u0cb0\u0cb5 \u0cb8\u0cbe\u0cb0\u0c9c\u0ca8\u0c95\u0c95\u0ccd\u0c95\u0cc6 \u0cb5\u0cb0\u0ccd\u0c97\u0cbe\u0caf\u0cbf\u0cb8\u0cbf.<\/p>\n

      \u0c89\u0caa\u0cb8\u0c82\u0cb8\u0ccd\u0c95\u0cc3\u0ca4\u0cbf \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0c98\u0ca8\u0cc0\u0c95\u0cb0\u0cbf\u0cb8\u0cc1\u0cb5 \u0cab\u0cc8\u0cac\u0ccd\u0cb0\u0cca\u0cac\u0ccd\u0cb2\u0cbe\u0cb8\u0ccd\u0c9f\u0ccd\u200c\u0c97\u0cb3\u0cbf\u0c97\u0cbe\u0c97\u0cbf \u0caa\u0ccd\u0cb0\u0ccb\u0c9f\u0ccb\u0c95\u0cbe\u0cb2\u0ccd<\/a><\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n

      [\/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||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 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 _builder_version=\u201d4.16\u2033 global_colors_info=\u201d{}\u201d]<\/p>\n

      \u0cb9\u0cc6\u0c9a\u0ccd\u0c9a\u0cbf\u0ca8 \u0cae\u0cbe\u0cb9\u0cbf\u0ca4\u0cbf\u0c97\u0cbe\u0c97\u0cbf \u0ca6\u0caf\u0cb5\u0cbf\u0c9f\u0ccd\u0c9f\u0cc1 \u0cb8\u0c82\u0caa\u0cb0\u0ccd\u0c95\u0cbf\u0cb8\u0cbf:<\/strong><\/h4>\n

      [\/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 locked=\u201doff\u201d global_colors_info=\u201d{}\u201d][et_pbner 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<\/p>\n

      \u0cb2\u0cc6\u0cb8\u0ccd\u0cb2\u0cbf B. \u0c97\u0cbe\u0cb0\u0ccd\u0ca1\u0ca8\u0ccd, MD, PhD<\/h6>\n

      \u0cac\u0ccd\u0cb0\u0ccc\u0ca8\u0ccd \u0cb5\u0cbf\u0cb6\u0ccd\u0cb5\u0cb5\u0cbf\u0ca6\u0ccd\u0caf\u0cbe\u0ca8\u0cbf\u0cb2\u0caf\u0ca6 \u0caa\u0cc0\u0ca1\u0cbf\u0caf\u0cbe\u0c9f\u0ccd\u0cb0\u0cbf\u0c95\u0ccd\u0cb8\u0ccd \u0cb8\u0c82\u0cb6\u0ccb\u0ca7\u0ca8\u0cc6\u0caf \u0caa\u0ccd\u0cb0\u0cbe\u0ca7\u0ccd\u0caf\u0cbe\u0caa\u0c95 \u0cb5\u0cbe\u0cb0\u0cc6\u0ca8\u0ccd \u0c86\u0cb2\u0ccd\u0caa\u0cb0\u0ccd\u0c9f\u0ccd \u0cb5\u0cc8\u0ca6\u0ccd\u0caf\u0c95\u0cc0\u0caf \u0cb6\u0cbe\u0cb2\u0cc6 \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0caa\u0cc0\u0ca1\u0cbf\u0caf\u0cbe\u0c9f\u0ccd\u0cb0\u0cbf\u0c95\u0ccd\u0cb8\u0ccd \u0cb5\u0cbf\u0cad\u0cbe\u0c97, \u0cb9\u0ccd\u0caf\u0cbe\u0cb8\u0ccd\u0cac\u0ccd\u0cb0\u0cca \u0cae\u0c95\u0ccd\u0c95\u0cb3 \u0c86\u0cb8\u0ccd\u0caa\u0ca4\u0ccd\u0cb0\u0cc6, \u0caa\u0ccd\u0cb0\u0cbe\u0cb5\u0cbf\u0ca1\u0cc6\u0ca8\u0ccd\u0cb8\u0ccd, \u0c85\u0cb0\u0cbf\u0cb5\u0cb3\u0cbf\u0c95\u0cc6 RI \u0cb5\u0cbf\u0cad\u0cbe\u0c97, \u0cae\u0c95\u0ccd\u0c95\u0cb3 \u0c86\u0cb8\u0ccd\u0caa\u0ca4\u0ccd\u0cb0\u0cc6 \u0cac\u0ccb\u0cb8\u0ccd\u0c9f\u0ca8\u0ccd \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0cb9\u0cbe\u0cb0\u0ccd\u0cb5\u0cb0\u0ccd\u0ca1\u0ccd \u0cb5\u0cc8\u0ca6\u0ccd\u0caf\u0c95\u0cc0\u0caf \u0cb6\u0cbe\u0cb2\u0cc6, \u0cac\u0ccb\u0cb8\u0ccd\u0c9f\u0ca8\u0ccd, MA \u0cb5\u0cc8\u0ca6\u0ccd\u0caf\u0c95\u0cc0\u0caf \u0ca8\u0cbf\u0cb0\u0ccd\u0ca6\u0cc7\u0cb6\u0c95, \u0caa\u0ccd\u0cb0\u0cca\u0c9c\u0cc6\u0cb0\u0cbf\u0caf\u0cbe \u0cb8\u0c82\u0cb6\u0ccb\u0ca7\u0ca8\u0cbe \u0caa\u0ccd\u0cb0\u0ca4\u0cbf\u0cb7\u0ccd\u0ca0\u0cbe\u0ca8<\/p>\n

      \u0ca6\u0cc2\u0cb0\u0cb5\u0cbe\u0ca3\u0cbf: 978-535-2594
      \u0cab\u0ccd\u0caf\u0cbe\u0c95\u0ccd\u0cb8\u0ccd: 508-543-0377
      <\/span>      lgordon@progeriaresearch.org<\/a><\/p>\n

      [\/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||=20pdpx\u201d \u0c95\u0cb8\u0ccd\u0c9f\u0cae\u0ccd 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|\u0ca1\u0cc6\u0cb8\u0ccd\u0c95\u0ccd\u200c\u0c9f\u0cbe\u0caa\u0ccd\u201d hover_enabled=\u201d0\u2033 global_colors_info=\u201d{}\u201d sticky_enabled=\u201d0\u2033]<\/p>\n

      \u0cb5\u0cc6\u0c82\u0ca1\u0cbf \u0ca8\u0cbe\u0cb0\u0ccd\u0cb0\u0cbf\u0cb8\u0ccd<\/span><\/h6>\n
      PRF \u0cb8\u0cc6\u0cb2\u0ccd \u0cae\u0ca4\u0ccd\u0ca4\u0cc1 \u0c9f\u0cbf\u0cb6\u0ccd\u0caf\u0cc2 \u0cac\u0ccd\u0caf\u0cbe\u0c82\u0c95\u0ccd
      \u0ca6\u0cc2\u0cb0\u0cb5\u0cbe\u0ca3\u0cbf: 401-274-1122 x 48063
      wnorris@brownhealth.org<\/a>
      <\/span><\/div>\n

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      \u0cb8\u0cc8\u0ca8\u0ccd \u0c85\u0caa\u0ccd \u0cae\u0cbe\u0ca1\u0cbf<\/h2>\n

      \u0ca8\u0cae\u0ccd\u0cae<\/h2>\n

      \u0cb8\u0cc1\u0ca6\u0ccd\u0ca6\u0cbf\u0caa\u0ca4\u0ccd\u0cb0!<\/h2>\n

      [\/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=\u201d\u0c88\u0c97 \u0ca6\u0cc7\u0ca3\u0cbf\u0c97\u0cc6 \u0ca8\u0cc0\u0ca1\u0cbf\u201d admin_label=\u201d\u0c92\u0c9f\u0ccd\u0c9f\u0cbf\u0c97\u0cc6, \u0ca8\u0cbe\u0cb5\u0cc1 \u0c9a\u0cbf\u0c95\u0cbf\u0ca4\u0ccd\u0cb8\u0cc6 \u0c95\u0c82\u0ca1\u0cc1\u0c95\u0cca\u0cb3\u0ccd\u0cb3\u0cc1\u0ca4\u0ccd\u0ca4\u0cc7\u0cb5\u0cc6! \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_T3\u201d2_slide 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\u201d0tabletz_0 border_radii=\u201don|25px|25px|25px|25px\u201d global_colors_info=\u201d{}\u201d button_bg_color__hover_enabled=\u201don\u201d button_bg_color__hover=\u201d#8fd2ed\u201d button_border_color__hover_enabled\u201d]<\/p>\n

      \u0c92\u0c9f\u0ccd\u0c9f\u0cbf\u0c97\u0cc6, \u0ca8\u0cbe\u0cb5\u0cc1<\/h2>\n

      \u0ca4\u0cbf\u0ca8\u0ccd\u0ca8\u0cc1\u0cb5\u0cc6<\/em><\/h2>\n

      \u0c9a\u0cbf\u0c95\u0cbf\u0ca4\u0ccd\u0cb8\u0cc6 \u0cb9\u0cc1\u0ca1\u0cc1\u0c95\u0cbf!<\/h2>\n

      [\/et_pb_cta][\/et_pb_column][et_pb_column type=\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 \u0cb8\u0ccd\u0c9f\u0ccd\u0cb0\u0cbf\u0caa\u0ccd \u0c85\u0ca1\u0cbf\u0c9f\u0cbf\u0caa\u0ccd\u0caa\u0ca3\u0cbf \u0caa\u0c9f\u0ccd\u0c9f\u0cbf\u0caf \u0ca8\u0c95\u0cb2\u0cc1 _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_ction<\/p>","protected":false},"excerpt":{"rendered":"

      [\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=\"Fibroblast Cell Culture Protocols\" font_container=\"tag:h1|text_align:center\" use_theme_fonts=\"yes\"][vc_column_text]\r\n

      PROTOCOL FOR SUBCULTURING AND FREEZING FIBROBLASTS\u00a0<\/strong><\/p>\r\n[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column width=\"1\/2\"][vc_column_text]Growth Media<\/u><\/strong>\r\n\r\nDMEM - ThermoFisher #11960-044 (high glucose without L-glutamine)\r\n\r\n15% FBS Fetal Bovine Serum \u2013 ThermoFisher #10437-028\r\n\r\n1% (1X) Penicillin-Streptomycin - ThermoFisher #15140-122\r\n\r\n1% (1X) GlutaMAX \u2013 ThermoFisher #35050-061[\/vc_column_text][\/vc_column][vc_column width=\"1\/2\"][vc_column_text]Trypsin<\/u><\/strong>\r\n\r\nTrypsin EDTA C 0.25% (ThermoFisher #25200-056)\r\n\r\nHank\u2019s Balanced Salt Solution\r\n<\/u><\/strong>HBSS- (ThermoFisher #14170 (1X) (-) calcium chloride, (-) magnesium chloride, (-) magnesium sulfate\r\n\r\nDMSO for Freezing\r\n<\/u><\/strong>Cell Culture Grade DMSO - Sigma #D2438[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]Thawing PRF cell lines<\/u><\/strong>\r\n

        \r\n \t
      1. Thaw cells rapidly in a 37\u02daC water bath.<\/li>\r\n \t
      2. Wipe the outside of the vial with 70% ethanol.<\/li>\r\n \t
      3. Transfer the thawed cells to a T25 flask containing 5 ml of growth media.<\/li>\r\n \t
      4. Place flask in 37\u02daC incubator overnight.<\/li>\r\n \t
      5. The following day, observe under the microscope to ensure cells have attached.<\/li>\r\n \t
      6. Remove media and replace with fresh growth media.<\/li>\r\n<\/ol>\r\n[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]Subculturing PRF cell lines<\/u><\/strong>\r\n
          \r\n \t
        1. Cells should be split when confluent.<\/li>\r\n \t
        2. To split cells (volumes given are assuming cells are in a T25):\r\na)\u00a0 Using sterile technique, remove media and rinse flask with 2-3 ml of sterile HBSS. Remove and discard HBSS.\r\nb)\u00a0 Add 1 ml trypsin and incubate for 2-3 minutes. Cells should be checked under an inverted microscope to determine if they have begun to round up, lift and float.\u00a0 If cells are not detached after 3 minutes, you may incubate another 1-2 minutes.\r\nc)\u00a0 Tighten cap and gently tip flask from side to side to dislodge all cells. Flask may be tapped lightly on side to loosen cells if needed.\r\nd)\u00a0 Add 4 ml of new media to flask as soon as cells are floating to inactivate the trypsin. Rinse down sides of flask several times with the new media to wash all cells off the plastic and into the solution. Remove all liquid containing cells and transfer to a 15ml conical (or 50 ml if you are pooling 3 or more flasks).\r\ne)\u00a0 Using sterile technique, remove an aliquot for counting on a hemocytometer.\r\nf)\u00a0 While you are counting cells, the rest of the solution should be spun in the clinical centrifuge for 5 minutes at 1000 rpm.<\/li>\r\n \t
        3. After calculating your cell count, plate cells at 2.5 x 105<\/sup> cells per T25 filter top flask.<\/li>\r\n \t
        4. Cells should be fed every 2-3 days with fresh growth medium.<\/li>\r\n<\/ol>\r\n[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]Freezing:\r\n<\/u><\/strong>Cells should be frozen at no less than 5 x 105<\/sup> cells\/ml\/cryovial in growth media containing 10% DMSO and 30% FBS and subsequently placed in an isopropanol freezing chamber at -80\u02daC overnight.\u00a0 Transfer to the liquid nitrogen the next day.\r\n\r\n\"\"\u00a0Protocol for Subculturing and Freezing Fibroblasts<\/a>[\/vc_column_text][\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]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\nLeslie_Gordon@brown.edu\u00a0<\/a>\r\n
          Wendy Norris<\/span><\/div>\r\n
          PRF Cell and Tissue Bank\r\nPhone: 401-274-1122 x 48063\r\nwnorris@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-1006","page","type-page","status-publish","hentry"],"yoast_head":"\nFibroblast | The Progeria Research Foundation<\/title>\n<meta name=\"description\" content=\"For info regarding Fibroblast Cell Culture Protocols, please contact Leslie Gordon at Leslie_Gordon@brown.edu, or Wendy Norris at wnorris@lifespan.org.\" \/>\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\/kn\/fibroblast-cell-culture-protocols\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Fibroblast | The Progeria Research Foundation\" 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