{"id":1008,"date":"2017-02-27T21:23:43","date_gmt":"2017-02-27T21:23:43","guid":{"rendered":"https:\/\/www.progeriaresearch.org\/?page_id=1008"},"modified":"2024-10-23T11:04:58","modified_gmt":"2024-10-23T15:04:58","slug":"immortalized-cell-culture-protocols","status":"publish","type":"page","link":"https:\/\/www.progeriaresearch.org\/ko\/immortalized-cell-culture-protocols\/","title":{"rendered":"\ubd88\uba78\ud654\ub41c \uc138\ud3ec \ubc30\uc591 \ud504\ub85c\ud1a0\ucf5c"},"content":{"rendered":"

[et_pb_section fb_built="1" \uc804\uccb4 \ub108\ube44="\ucf1c\uc9d0" \ube44\ud65c\uc131\ud654\ub428_\ucf1c\uc9d0="\uaebc\uc9d0|\uaebc\uc9d0|\uaebc\uc9d0" _\ube4c\ub354_\ubc84\uc804="4.16" \ud14c\ub450\ub9ac_\ub108\ube44_\ubc14\ub2e5="55px" \ud14c\ub450\ub9ac_\uc0c9\uc0c1_\ubc14\ub2e5="#29327a" \uc7a0\uae40="\uaebc\uc9d0" \uae00\ub85c\ubc8c_\uc0c9\uc0c1_\uc815\ubcf4="{}"][et_pb_\uc804\uccb4 \ub108\ube44_\ud5e4\ub354 _\ube4c\ub354_\ubc84\uc804="4.16" \uc81c\ubaa9_\uae00\uaf34="|||||||||" \uc81c\ubaa9_\uae00\uaf34_\ud06c\uae30="55" \ubc30\uacbd_\uc0c9\uc0c1="#29327a" \ubc30\uacbd_\uc774\ubbf8\uc9c0="https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2020\/12\/Aluisio.jpg" \ubc30\uacbd_\uc704\uce58="center_left" \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529="9vw||9vw||true" \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529_\ud0dc\ube14\ub9bf="" \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529_\ud734\ub300\ud3f0="|56px||" \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529_\ub9c8\uc9c0\ub9c9_\ud3b8\uc9d1="\ub370\uc2a4\ud06c\ud1b1\uc5d0\uc11c" \uc81c\ubaa9_\uae00\uaf34_\ud06c\uae30_\ud0dc\ube14\ub9bf="45px" \uc81c\ubaa9_\uae00\uaf34_\ud06c\uae30_\ud734\ub300\ud3f0="40px" \uc81c\ubaa9_\uae00\uaf34_\ud06c\uae30_\ub9c8\uc9c0\ub9c9_\ud3b8\uc9d1="\ud734\ub300\ud3f0\uc5d0\uc11c" z_index_tablet="500" \uc0ac\uc6a9\uc790 \uc815\uc758_css_main_element="\ubc30\uacbd-\uc704\uce58: \uac00\uc6b4\ub370 18% !\uc911\uc694;" global_colors_info="{}"]<\/p>\n

\ubd88\uba78\ud654\ub41c \uc138\ud3ec<\/h1>\n

\ubb38\ud654 \ud504\ub85c\ud1a0\ucf5c<\/h1>\n

 <\/p>\n

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\n[\/et_pb_sidebar][\/et_pb_column][et_pb_column type="3_4" specialty_columns="3" _builder_version="4.16" custom_padding="|||" global_colors_info="{}" custom_padding__hover="|||"][et_pb_row_inner _builder_version="4.16" background_color="#00b2e2" custom_margin="41px||" custom_margin_tablet="0px||" custom_margin_phone="" custom_margin_last_edited="on|phone" custom_padding="39.4375px|20px|0|20px|false|true" animation_style="slide" animation_direction="top" animation_intensity_slide="25%" box_shadow_style="preset1" box_shadow_blur="38px" box_shadow_spread="-12px" locked="off" global_colors_info="{}"][et_pb_column_inner saved_specialty_column_type="3_4" _builder_version="4.16" custom_padding="|||" global_colors_info="{}" custom_padding__hover="|||"][et_pb_text admin_label="\ud504\ub85c\uac8c\ub9ac\uc544 \uc5f0\uad6c \uc7ac\ub2e8\uc758 \uc791\uc5c5..." _builder_version="4.16" header_2_line_height="1.2em" background_layout="dark" custom_margin="||35px" custom_padding="|||" custom_padding_tablet="|50px||50px||true" custom_padding_phone="" custom_padding_last_edited="on|\ub370\uc2a4\ud06c\ud0d1" header_font_size_tablet="" header_font_size_phone="" header_font_size_last_edited="on|\ub370\uc2a4\ud06c\ud0d1" header_2_font_size_tablet="25px" header_2_font_size_phone="" header_2_font_size_last_edited="on|\ud0dc\ube14\ub9bf" header_4_font_size_tablet="" header_4_font_size_phone="19px" header_4_font_size_last_edited="on|\ud734\ub300\ud3f0" header_4_line_height_tablet="" header_4_line_height_phone="1.2em" header_4_line_height_last_edited="on|\ud734\ub300\ud3f0" z_index_tablet="500" \uae00\ub85c\ubc8c_\uc0c9\uc0c1_\uc815\ubcf4="{}"]<\/p>\n

\ubd88\uba78\ud654\ub41c \uc138\ud3ec \ubc30\uc591 \ud504\ub85c\ud1a0\ucf5c<\/h2>\n

[\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner \ucee4\uc2a4\ud140 \ud328\ub529_\ub9c8\uc9c0\ub9c9_\ud3b8\uc9d1="on|\ud734\ub300\ud3f0" _\ube4c\ub354_\ubc84\uc804="4.16" \ucee4\uc2a4\ud140 \ud328\ub529="||0px|||" \ucee4\uc2a4\ud140 \ud328\ub529_\ud0dc\ube14\ub9bf="|35px||35px||true" \ucee4\uc2a4\ud140 \ud328\ub529_\ud734\ub300\ud3f0="" \ud14c\ub450\ub9ac_\ub108\ube44_\uc704\ucabd="10px" \ud14c\ub450\ub9ac_\uc0c9\uc0c1_\uc704\ucabd="#8fd2ed" \uae00\ub85c\ubc8c_\uc0c9\uc0c1_\uc815\ubcf4="{}"][et_pb_column_inner \uc800\uc7a5\ub41c_\ud2b9\uc218_\uc5f4_\uc720\ud615="3_4" _\ube4c\ub354_\ubc84\uc804="4.16" \ucee4\uc2a4\ud140 \ud328\ub529="|||" global_colors_info="{}" \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529__\ud638\ubc84="|||"][et_pb_text _builder_version="4.16" background_size="initial" background_position="top_left" background_repeat="repeat" \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529="||0px|||" global_colors_info="{}"]<\/p>\n

\ud558\ubc84\ub4dc \ub300\ud559\uc758 \ub9c8\ud2f4 \ub3c4\ub974\ud504 \ubc15\uc0ac\uac00 PRF \uc138\ud3ec \ubc0f \uc870\uc9c1 \uc740\ud589\uc5d0 \ubd88\uba78\ud654\ub41c \uc138\ud3ec\uc8fc\ub97c \uc0dd\uc131\ud558\uc5ec \uc81c\uacf5\ud574 \uc8fc\uc2e0 \ub370 \ub300\ud574 \uac10\uc0ac\ub4dc\ub9bd\ub2c8\ub2e4. \uac10\uc0ac\ud569\ub2c8\ub2e4.<\/p>\n

PRF \uc138\ud3ec\uc8fc\ub294 \ub2e4\uc74c \ud504\ub85c\ud1a0\ucf5c\uc744 \uc0ac\uc6a9\ud558\uc5ec \ubd88\uba78\ud654\ub418\uc5c8\uc2b5\ub2c8\ub2e4.<\/p>\n

\ub808\ud2b8\ub85c\ubc14\uc774\ub7ec\uc2a4 \uac10\uc5fc<\/strong><\/h4>\n

A. \ubc14\uc774\ub7ec\uc2a4 \uc0dd\uc131:<\/strong> \ud615\uc9c8\uac10\uc5fc 12-24\uc2dc\uac04 \uc804\uc5d0 HEK293 \ud328\ud0a4\uc9d5 \uc138\ud3ec\ub97c 60-80% \ucee8\ud50c\ub8e8\uc5b8\uc2dc\uc5d0\uc11c 100mm \ud50c\ub808\uc774\ud2b8\uc5d0 \ub3c4\ub9d0\ud588\uc2b5\ub2c8\ub2e4. \uac01 \ud50c\ub808\uc774\ud2b8\ub294 12\u03bcg\uc758 \uc5ed\uc804\uc0ac \ubc14\uc774\ub7ec\uc2a4 \uad6c\uc131\uccb4 pBABE-puro(SV40T)\uc640 12\u03bcg\uc758 pCL-Ampho \uc5ed\uc804\uc0ac \ubc14\uc774\ub7ec\uc2a4 \ud328\ud0a4\uc9d5 \ubca1\ud130\ub85c \uacf5\ub3d9 \ud615\uc9c8\uac10\uc5fc\ub418\uc5c8\uc2b5\ub2c8\ub2e4. \ud615\uc9c8\uac10\uc5fc 8-10\uc2dc\uac04 \ud6c4\uc5d0 \ubc30\uc591 \ubc30\uc9c0\ub97c \ud761\uc778\ud558\uace0 10ml\uc758 \uc644\uc804 \ubc30\uc9c0\ub85c \uad50\uccb4\ud588\uc2b5\ub2c8\ub2e4. \ubc30\uc591\ubb3c\uc744 \ucd94\uac00\ub85c 48-72\uc2dc\uac04 \ub3d9\uc548 \ubc30\uc591\ud558\uc5ec \ucd5c\uc801\uc758 \ubc14\uc774\ub7ec\uc2a4 \uc5ed\uac00\ub97c \uc5bb\uc5c8\uc2b5\ub2c8\ub2e4.<\/p>\n

B. \ud45c\uc801 \uc138\ud3ec \uac10\uc5fc:<\/strong> \ud45c\uc801 \uc138\ud3ec\ub294 \uac10\uc5fc 12-18\uc2dc\uac04 \uc804\uc5d0 100mm \ud50c\ub808\uc774\ud2b8\uc5d0 \ub3c4\ub9d0\ud588\uc2b5\ub2c8\ub2e4. \ud3ec\uc7a5 \uc138\ud3ec\uc5d0\uc11c \ubc30\uc9c0\ub97c \uc218\uc9d1\ud558\uc5ec 0.45-\u03bcm \uc140\ub8f0\ub85c\uc2a4 \uc544\uc138\ud14c\uc774\ud2b8 \ub610\ub294 \ud3f4\ub9ac\uc124\ud3f0 \ud544\ud130\ub85c \uc5ec\uacfc\ud588\uc2b5\ub2c8\ub2e4. \uc138\ud3ec\uac00 40-60% \ud569\ub958\ub418\uba74 \ucd5c\uc885 \ub18d\ub3c4\uac00 10\u03bcg\/ml \ud3f4\ub9ac\ube0c\ub80c\uc778 8ml \ubc14\uc774\ub7ec\uc2a4 \ud568\uc720 \ubc30\uc9c0\ub97c \ucca8\uac00\ud588\uc2b5\ub2c8\ub2e4. 3\ud68c \uac10\uc5fc\uc744 \uc5f0\uc18d\uc801\uc73c\ub85c \uc218\ud589\ud588\uc73c\uba70, \uc57d 12\uc2dc\uac04 \uac04\uaca9\uc744 \ub450\uc5c8\uc2b5\ub2c8\ub2e4. \ubc30\uc9c0\ub294 24\uc2dc\uac04 \ubc30\uc591 \ud6c4 \uad50\uccb4\ud588\uc2b5\ub2c8\ub2e4.<\/p>\n

C. \uc548\uc815\ub41c \uc138\ud3ec\uc8fc \uc120\ud0dd:<\/strong> \uac10\uc5fc 72\uc2dc\uac04 \ud6c4, \uc138\ud3ec\ub97c \ud2b8\ub9bd\uc2e0 \ucc98\ub9ac\ud558\uace0 1:3\uc73c\ub85c \ubd84\ud560\ud558\uc5ec 3 \u03bcg\/ml \ud478\ub85c\ub9c8\uc774\uc2e0\uacfc \ud56d\uc0dd\uc81c\uac00 \uc5c6\ub294 \ub300\uc870\uad70 \ud50c\ub808\uc774\ud2b8 1\uac1c\uac00 \uc788\ub294 \ub450 \uac1c\uc758 100 mm \ud50c\ub808\uc774\ud2b8\ub85c \uc62e\uacbc\uc2b5\ub2c8\ub2e4. \ubc30\uc591 \ubc30\uc9c0\ub294 \uc57d 2~4\uc8fc \ub3d9\uc548 2~3\uc77c\ub9c8\ub2e4 \uad50\uccb4\ud558\uc5ec \uc548\uc815\uc801\uc778 \uc138\ud3ec\uc8fc\ub97c \uc0dd\uc0b0\ud588\uc2b5\ub2c8\ub2e4.<\/p>\n

 <\/p>\n

[\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner column_structure="1_2,1_2" custom_padding_last_edited="on|phone" _builder_version="4.16" custom_margin="|||||" custom_padding="0px|35px|0|0px|false|false" custom_padding_tablet="|35px|20px|35px||true" custom_padding_phone="|35px|11px|35px|false|true" animation_direction="top" global_colors_info="{}"][et_pb_column_inner type="1_2" saved_specialty_column_type="3_4" _builder_version="4.16" custom_padding="|20px||" custom_padding_tablet="||20px" custom_padding_phone="" custom_padding_last_edited="on|desktop" global_colors_info="{}" custom_padding__hover="|||"][et_pb_text _builder_version="4.16" background_size="initial" background_position="top_left" background_repeat="repeat" custom_padding="||24px|||" global_colors_info="{}"]<\/p>\n

\uc131\uc7a5 \ub9e4\uccb4<\/h4>\n

DMEM- Invitrogen #11960-044(L-\uae00\ub8e8\ud0c0\ubbfc\uc774 \uc5c6\ub294 \uace0\ub18d\ub3c4 \ud3ec\ub3c4\ub2f9)<\/span><\/p>\n

15% FBS \ud0dc\uc544\uc6b0\ud608\uccad<\/span><\/p>\n

1%(1x) \ud398\ub2c8\uc2e4\ub9b0-\uc2a4\ud2b8\ub819\ud1a0\ub9c8\uc774\uc2e0(Invitrogen#15140-122)<\/span><\/p>\n

1%(1X) L-\uae00\ub8e8\ud0c0\ubbfc(Invitrogen#25030-081)<\/span><\/p>\n

0.75 \u03bcg\/ml \ud478\ub85c\ub9c8\uc774\uc2e0(InvivoGen #ant-pr-1)<\/span><\/p>\n

[\/et_pb_text][\/et_pb_column_inner][et_pb_column_inner type="1_2" saved_specialty_column_type="3_4" _builder_version="4.16" custom_padding="|||20px" custom_padding_tablet="20px|||0px" custom_padding_phone="" custom_padding_last_edited="on|phone" global_colors_info="{}" custom_padding__hover="|||"][et_pb_text _builder_version="4.16" background_size="initial" background_position="top_left" background_repeat="repeat" custom_padding="||24px|||" global_colors_info="{}"]<\/p>\n

\ud2b8\ub9bd\uc2e0<\/strong><\/h4>\n

\ud2b8\ub9bd\uc2e0 EDTA C .25%(\uc778\ube44\ud2b8\ub85c\uac90#25200-056)<\/span><\/p>\n

\ud578\ud06c\uc758 \uade0\ud615 \uc18c\uae08 \uc194\ub8e8\uc158<\/strong><\/h4>\n

HBSS- (Invitrogen#14170 (1X) (-) \uc5fc\ud654\uce7c\uc298, (-) \uc5fc\ud654\ub9c8\uadf8\ub124\uc298, (-) \ud669\uc0b0\ub9c8\uadf8\ub124\uc298)<\/span><\/p>\n

\ub3d9\uacb0\uc744 \uc704\ud55c DMSO<\/strong><\/h4>\n

\uc138\ud3ec \ubc30\uc591 \ub4f1\uae09 DMSO<\/span><\/p>\n

[\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner \ucee4\uc2a4\ud140 \ud328\ub529_\ub9c8\uc9c0\ub9c9_\ud3b8\uc9d1="on|\ud734\ub300\ud3f0" _\ube4c\ub354_\ubc84\uc804="4.16" \ucee4\uc2a4\ud140_\ub9c8\uc9c4="15px||35px|||" \ucee4\uc2a4\ud140_\ud328\ub529="0px|35px|0|0px|false|false" \ucee4\uc2a4\ud140_\ud328\ub529_\ud14c\uc774\ube14="|35px|20px|35px||true" \ucee4\uc2a4\ud140_\ud328\ub529_\ud3f0="|35px|11px|35px|false|true" \uc560\ub2c8\uba54\uc774\uc158_\ubc29\ud5a5="\uc704" \uae00\ub85c\ubc8c_\uc0c9\uc0c1_\uc815\ubcf4="{}"][et_pb_column_inner \uc800\uc7a5\ub41c_\ud2b9\uc218_\uc5f4_\uc720\ud615="3_4" _\ube4c\ub354_\ubc84\uc804="4.16" \ucee4\uc2a4\ud140_\ud328\ub529="|20px||" custom_padding_tablet="||20px" custom_padding_phone="" custom_padding_last_edited="on|desktop" global_colors_info="{}" custom_padding__hover="|||"][et_pb_text _builder_version="4.16" custom_margin="||35px" global_colors_info="{}"]<\/p>\n

\uc138\ud3ec\ub294 \uc57d 5 x 10\uc758 \ub18d\ub3c4\ub85c \uac74\uc870 \uc5bc\uc74c \uc704\uc5d0 \ub3d9\uacb0\ub418\uc5b4 \ub3c4\ucc29\ud569\ub2c8\ub2e4.5<\/sup>\uc138\ud3ec\/\ubc14\uc774\uc54c<\/b><\/p>\n

[\/et_pb_text][et_pb_text _builder_version="4.16" background_size="initial" background_position="top_left" background_repeat="repeat" custom_padding="||0px|||" global_colors_info="{}"]<\/p>\n

\uc12c\uc720\uc544\uc138\ud3ec \ud574\ub3d9 \ud504\ub85c\ud1a0\ucf5c<\/b><\/h4>\n
    \n
  1. 37\u00b0C \uc218\uc870\uc5d0\uc11c \uc138\ud3ec\ub97c \ube60\ub974\uac8c \ub179\uc785\ub2c8\ub2e4.<\/li>\n
  2. 70% \uc5d0\ud0c4\uc62c\ub85c \ubc14\uc774\uc54c \uc678\ubd80\ub97c \ub2e6\uc2b5\ub2c8\ub2e4.<\/li>\n
  3. \ud574\ub3d9\ub41c \uc138\ud3ec\ub97c \ud4e8\ub85c\ub9c8\uc774\uc2e0\uc774 \uc5c6\ub294 5ml\uc758 \uc131\uc7a5 \ubc30\uc9c0\uac00 \ub4e4\uc5b4 \uc788\ub294 T25 \ud50c\ub77c\uc2a4\ud06c\ub85c \uc62e\uae41\ub2c8\ub2e4.<\/li>\n
  4. \ud50c\ub77c\uc2a4\ud06c\ub97c 37\u00b0C \uc778\ud050\ubca0\uc774\ud130\uc5d0 \ubc24\uc0c8 \ub123\uc5b4\ub461\ub2c8\ub2e4.<\/li>\n
  5. \ub2e4\uc74c \ub0a0, \ud604\ubbf8\uacbd\uc73c\ub85c \uad00\ucc30\ud558\uc5ec \uc138\ud3ec\uac00 \ubd80\ucc29\ub418\uc5c8\ub294\uc9c0 \ud655\uc778\ud558\uc138\uc694.<\/li>\n
  6. \ubc30\uc9c0\ub97c \uc81c\uac70\ud558\uace0 \uc0c8\ub85c\uc6b4 \uc131\uc7a5 \ubc30\uc9c0\ub85c \uad50\uccb4\ud569\ub2c8\ub2e4.<\/li>\n<\/ol>\n

    PRF \uc138\ud3ec\uc8fc \ud558\uc704 \ubc30\uc591<\/b><\/h4>\n

    1) \uc138\ud3ec\uac00 \ud655\ub9bd\ub418\uace0 \uc131\uc7a5\ud558\uba74 0.75 \u03bcg\/ml puromycin\uc774 \ud3ec\ud568\ub41c \ubc30\uc9c0\ub97c \uc0ac\uc6a9\ud558\uae30 \uc2dc\uc791\ud569\ub2c8\ub2e4. 0.75 \u03bcg\/ml puromycin\uc774 \ud3ec\ud568\ub41c \ubc30\uc9c0\uc5d0\uc11c \uc138\ud3ec\ub97c \uacc4\uc18d \uc720\uc9c0\ud558\uc2ed\uc2dc\uc624.<\/p>\n

    2) \uc138\ud3ec\ub294 \ud569\ub958\ud560 \ub54c \ubd84\ud560\ub418\uc5b4\uc57c \ud569\ub2c8\ub2e4.<\/p>\n

    3) \uc140 \ubd84\ud560(\uc8fc\uc5b4\uc9c4 \ubcfc\ub968\uc740 \uc140\uc774 T25\uc5d0 \uc788\ub2e4\uace0 \uac00\uc815\ud568):<\/p>\n

    A. \uba78\uade0 \uae30\uc220\uc744 \uc0ac\uc6a9\ud558\uc5ec \ubc30\uc9c0\ub97c \uc81c\uac70\ud558\uace0 2-3ml\uc758 \uba78\uade0 HBSS\ub85c \ud50c\ub77c\uc2a4\ud06c\ub97c \ud5f9\uad7d\ub2c8\ub2e4. HBSS\ub97c \uc81c\uac70\ud558\uace0 \ud3d0\uae30\ud569\ub2c8\ub2e4.<\/p>\n

    B. \ud2b8\ub9bd\uc2e0 1ml\ub97c \ub123\uace0 2-3\ubd84\uac04 \ubc30\uc591\ud569\ub2c8\ub2e4. \uc138\ud3ec\ub97c \uc5ed\ud604\ubbf8\uacbd\uc73c\ub85c \uac80\uc0ac\ud558\uc5ec \ub465\uae00\uac8c \ubb49\uccd0\uc9c0\uace0, \ub4e4\uc5b4\uc62c\ub824\uc9c0\uace0, \ub5a0\ub2e4\ub2c8\uae30 \uc2dc\uc791\ud588\ub294\uc9c0 \ud655\uc778\ud574\uc57c \ud569\ub2c8\ub2e4. 3\ubd84 \ud6c4\uc5d0\ub3c4 \uc138\ud3ec\uac00 \ubd84\ub9ac\ub418\uc9c0 \uc54a\uc73c\uba74 1-2\ubd84 \ub354 \ubc30\uc591\ud560 \uc218 \uc788\uc2b5\ub2c8\ub2e4.<\/p>\n

    C. \ucea1\uc744 \uc870\uc774\uace0 \ud50c\ub77c\uc2a4\ud06c\ub97c \uc88c\uc6b0\ub85c \uc0b4\uc9dd \uae30\uc6b8\uc5ec \ubaa8\ub4e0 \uc138\ud3ec\ub97c \uc81c\uac70\ud569\ub2c8\ub2e4. \ud544\uc694\ud55c \uacbd\uc6b0 \ud50c\ub77c\uc2a4\ud06c\ub97c \uac00\ubccd\uac8c \ub450\ub4dc\ub824 \uc138\ud3ec\ub97c \ub290\uc2a8\ud558\uac8c \ud560 \uc218 \uc788\uc2b5\ub2c8\ub2e4.<\/p>\n

    D. \uc138\ud3ec\uac00 \ub5a0\ub2e4\ub2c8\ub294 \uc989\uc2dc \ud50c\ub77c\uc2a4\ud06c\uc5d0 \uc0c8\ub85c\uc6b4 \ubc30\uc9c0 4ml\ub97c \ub123\uc5b4 \ud2b8\ub9bd\uc2e0\uc744 \ubd88\ud65c\uc131\ud654\ud569\ub2c8\ub2e4. \ud50c\ub77c\uc2a4\ud06c \uce21\uba74\uc744 \uc0c8\ub85c\uc6b4 \ubc30\uc9c0\ub85c \uc5ec\ub7ec \ubc88 \ud5f9\uad6c\uc5b4 \ud50c\ub77c\uc2a4\ud2f1\uc5d0\uc11c \ubaa8\ub4e0 \uc138\ud3ec\ub97c \uc53b\uc5b4\ub0b4\uace0 \uc6a9\uc561\uc73c\ub85c \uc62e\uae41\ub2c8\ub2e4. \uc138\ud3ec\uac00 \ub4e4\uc5b4 \uc788\ub294 \ubaa8\ub4e0 \uc561\uccb4\ub97c \uc81c\uac70\ud558\uace0 15ml \uc6d0\ubfd4\ud615(\ub610\ub294 3\uac1c \uc774\uc0c1\uc758 \ud50c\ub77c\uc2a4\ud06c\ub97c \ubaa8\uc744 \uacbd\uc6b0 50ml)\uc73c\ub85c \uc62e\uae41\ub2c8\ub2e4.<\/p>\n

    E. \ubb34\uade0 \uae30\uc220\uc744 \uc0ac\uc6a9\ud558\uc5ec \ud608\uc561\uc744 \ucc44\ucde8\ud558\uc5ec \ud608\uad6c\uacc4\uc0b0\uae30\ub85c \uce21\uc815\ud569\ub2c8\ub2e4.<\/p>\n

    F. \uc138\ud3ec\ub97c \uc138\ub294 \ub3d9\uc548 \ub098\uba38\uc9c0 \uc6a9\uc561\uc740 \uc784\uc0c1\uc6a9 \uc6d0\uc2ec\ubd84\ub9ac\uae30\uc5d0\uc11c 1000rpm\uc73c\ub85c 5\ubd84 \ub3d9\uc548 \ud68c\uc804\uc2dc\ucf1c\uc57c \ud569\ub2c8\ub2e4.<\/p>\n

    4) \uc138\ud3ec \uc218\ub97c \uacc4\uc0b0\ud55c \ud6c4, \uc138\ud3ec\ub97c 2.5 x 105<\/sup>\u00a0T 25 \ud544\ud130 \uc0c1\ubd80 \ud50c\ub77c\uc2a4\ud06c\ub2f9 \uc138\ud3ec.<\/p>\n

    5) \uc138\ud3ec\uc5d0\ub294 0.75 \u03bcg\/ml\uc758 \ud4e8\ub85c\ub9c8\uc774\uc2e0\uc774 \ud568\uc720\ub41c \uc2e0\uc120\ud55c \uc131\uc7a5 \ubc30\uc9c0\ub97c 2~3\uc77c\ub9c8\ub2e4 \uacf5\uae09\ud574\uc57c \ud569\ub2c8\ub2e4.<\/p>\n

    \ub3d9\uacb0:<\/h4>\n

    <\/b>\uc138\ud3ec\ub294 \ucd5c\uc18c 5x 105<\/sup>\u00a0<\/span>\uc138\ud3ec \/ml\/cryovial\u00a0\uc131\uc7a5 \ub9e4\uccb4\uc5d0\uc11c<\/span>\u00a0<\/span>10% DMSO \ubc0f 30% FBS\ub97c \ud568\uc720\u00a0<\/span>\ud478\ub85c\ub9c8\uc774\uc2e0 \uc5c6\uc774<\/strong>\u00a0<\/span>\uadf8\ub9ac\uace0 \uadf8 \ud6c4 -80\u00b0C\uc758 \uc774\uc18c\ud504\ub85c\ud310\uc62c \ub0c9\ub3d9\uc2e4\uc5d0 \ubc24\uc0c8 \ub450\uc5c8\ub2e4. \ub2e4\uc74c \ub0a0 \uc561\uccb4 \uc9c8\uc18c\ub85c \uc62e\uae34\ub2e4.<\/p>\n

    [\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner \ucee4\uc2a4\ud140 \ud328\ub529_\ub9c8\uc9c0\ub9c9_\ud3b8\uc9d1="on|\ud734\ub300\ud3f0" _\ube4c\ub354_\ubc84\uc804="4.16" \ucee4\uc2a4\ud140 \ud328\ub529="||0px|||" \ucee4\uc2a4\ud140 \ud328\ub529_\ud0dc\ube14\ub9bf="|35px||35px||true" \ucee4\uc2a4\ud140 \ud328\ub529_\ud734\ub300\ud3f0="" \ud14c\ub450\ub9ac_\ub108\ube44_\uc704\ucabd="10px" \ud14c\ub450\ub9ac_\uc0c9\uc0c1_\uc704\ucabd="#8fd2ed" \uc7a0\uae08="\uaebc\uc9d0" \uae00\ub85c\ubc8c_\uc0c9\uc0c1_\uc815\ubcf4="{}"][et_pb_column_inner \uc800\uc7a5\ub41c_\ud2b9\uc218_\uc5f4_\uc720\ud615="3_4" _\ube4c\ub354_\ubc84\uc804="4.16" \ucee4\uc2a4\ud140 \ud328\ub529="|||" global_colors_info=\u201d{}\u201d \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529__\ud638\ubc84=\u201d|||\u201d][et_pb_text _\ube4c\ub354_\ubc84\uc804=\u201d4.16\u2033 global_colors_info=\u201d{}\u201d]<\/p>\n

    \uc790\uc138\ud55c \ub0b4\uc6a9\uc740 \ub2e4\uc74c \uc8fc\uc18c\ub85c \ubb38\uc758\ud558\uc138\uc694.<\/strong><\/h4>\n

    [\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row_inner column_structure="1_2,1_2" custom_padding_last_edited="on|phone" _builder_version="4.16" custom_padding="|35px|0|0px|false|false" custom_padding_tablet="|35px||35px||true" custom_padding_phone="" animation_direction="top" locked="off" global_colors_info="{}"][et_pb_column_inner type="1_2" saved_specialty_column_type="3_4" _builder_version="4.16" custom_padding="|20px||" custom_padding_tablet="||20px" custom_padding_phone="" custom_padding_last_edited="on|desktop" global_colors_info="{}" custom_padding__hover="|||"][et_pb_text _builder_version="4.19.5" global_colors_info="{}"]<\/p>\n

    Leslie B. Gordon, MD, PhD<\/h6>\n

    \ube0c\ub77c\uc6b4 \ub300\ud559\uad50\uc758 Warren Alpert \uc758\ub300 \uc18c\uc544\uacfc \uc5f0\uad6c \uad50\uc218, \ub85c\ub4dc\uc544\uc77c\ub79c\ub4dc \uc8fc \ud504\ub85c\ube44\ub358\uc2a4\uc5d0 \uc788\ub294 Hasbro \uc5b4\ub9b0\uc774 \ubcd1\uc6d0 \uc18c\uc544\uacfc, \ubcf4\uc2a4\ud134 \uc5b4\ub9b0\uc774 \ubcd1\uc6d0 \ub9c8\ucde8\uacfc, \ub9e4\uc0ac\ucd94\uc138\uce20 \uc8fc \ubcf4\uc2a4\ud134\uc5d0 \uc788\ub294 Harvard \uc758\ub300, Progeria Research Foundation \uc758\ub8cc \ucc45\uc784\uc790<\/p>\n

    \uc804\ud654: 978-535-2594
    \ud329\uc2a4: 508-543-0377
    <\/span>lgordon@progeriaresearch.org<\/a><\/p>\n

    [\/et_pb_text][\/et_pb_column_inner][et_pb_column_inner type="1_2" saved_specialty_column_type="3_4" _builder_version="4.16" custom_padding="|||20px" custom_padding_tablet="20px|||0px" custom_padding_phone="" custom_padding_last_edited="on|phone" global_colors_info="{}" custom_padding__hover="|||"][et_pb_text _builder_version="4.27.2" custom_padding="||0px|||" custom_padding_last_edited="off|desktop" \ud638\ubc84 \ud65c\uc131\ud654="0" global_colors_info="{}" sticky_enabled="0"]<\/p>\n

    \uc6ec\ub514 \ub178\ub9ac\uc2a4<\/span><\/h6>\n
    PRF \uc138\ud3ec \ubc0f \uc870\uc9c1 \uc740\ud589
    \uc804\ud654: 401-274-1122 x 48063
    wnorris@brownhealth.org<\/a><\/a><\/span><\/div>\n

    [\/et_pb_text][\/et_pb_column_inner][\/et_pb_row_inner][\/et_pb_column][\/et_pb_section][et_pb_section fb_built="1" module_class="footer" _builder_version="4.21.0" background_color="#29327a" custom_margin="-2px|||||" custom_padding="0|0px|0|0px|false|false" z_index_tablet="500" border_width_top="12px" border_color_top="#00b2e2" global_module="133" locked="off" global_colors_info="{}"][et_pb_row_inner column_structure="1_3,1_3,1_3" disabled_on="on|on|on" admin_label="\ud589" _builder_version="4.21.0" _module_preset="default" min_height="277px" custom_margin="|20px|30px||false|false" custom_padding="0px|30px|0px||false|false" disabled="on" global_colors_info="{}"][et_pb_column_inner type="1_3" saved_specialty_column_type="3_4" _builder_version="4.21.0" _module_preset="\uae30\ubcf8\uac12" global_colors_info="{}"][et_pb_image src="https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2023\/08\/Startbanner-1.png" title_text="Startbanner" url="https:\/\/www.gravoc.com\/" align="right" _builder_version="4.21.0" _module_preset="\uae30\ubcf8\uac12" custom_margin="|-50px||20px|false|false" global_colors_info="{}"][\/et_pb_image][\/et_pb_column_inner][et_pb_column_inner type="1_3" saved_specialty_column_type="3_4" _builder_version="4.21.0" _module_preset="default" global_colors_info="{}"][\/et_pb_column_inner][et_pb_column_inner type="1_3" saved_specialty_column_type="3_4" _builder_version="4.21.0" _module_preset="default" global_colors_info="{}"][et_pb_image src="https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2023\/07\/RFRFinish.png" title_text="RFRFinish" _builder_version="4.21.0" _module_preset="\uae30\ubcf8" custom_margin="|20px||-50px|false|false" global_colors_info="{}"][\/et_pb_image][\/et_pb_column_inner][\/et_pb_row_inner][et_pb_row column_structure="1_4,1_4,1_2" make_equal="on" module_class=" et_pb_row_fullwidth" _builder_version="4.16" width="89%" width_tablet="80%" width_phone="" width_last_edited="on|desktop" max_width="89%" max_width_tablet="80%" max_width_phone="" max_width_last_edited="on|\ub370\uc2a4\ud06c\ud1b1" z_index_tablet="500" make_fullwidth="on" width_unit="off" custom_width_percent="100%" global_colors_info="{}"][et_pb_column type="1_4" _builder_version="4.16" custom_padding="|||" global_colors_info="{}" custom_padding__hover="|||"][et_pb_cta button_url="https:\/\/www.progeriaresearch.org\/newsletter-signup\/" button_text="\uc9c0\uae08 \uac00\uc785\ud558\uc138\uc694" admin_label="\ub274\uc2a4\ub808\ud130 \uac00\uc785" module_class="sign-btn" _builder_version="4.16" header_font_size="25px" background_color="#29327a" animation_style="slide" animation_direction="left" animation_intensity_slide="25%" header_font_size_tablet="" header_font_size_phone="30px" header_font_size_last_edited="on|desktop" z_index_tablet="500" border_radii="on|25px|25px|25px|25px" global_colors_info="{}" button_bg_color__hover_enabled="on" button_bg_color__hover="#8fd2ed" button_border_color__hover_enabled="on"]<\/p>\n

    \uac00\uc785\ud558\uae30<\/h2>\n

    \uc6b0\ub9ac\ub97c \uc704\ud574<\/h2>\n

    \ud68c\ub78c \uc2e0\ubb38!<\/h2>\n

    [\/et_pb_cta][\/et_pb_column][et_pb_column type="1_4" _builder_version="4.16" custom_padding="|||" global_colors_info="{}" custom_padding__hover="|||"][et_pb_cta button_url="https:\/\/progeriaresearch.donorsupport.co\/-\/XZHJVWZR" button_text="\uc9c0\uae08 \uae30\ubd80\ud558\uae30" admin_label="\ud568\uaed8\ub77c\uba74 \uc6b0\ub9ac\ub294 \uce58\ub8cc\ubc95\uc744 \ucc3e\uc744 \uac83\uc785\ub2c8\ub2e4!" module_class="sign-btn" _builder_version="4.16" header_font_size="25px" background_color="#29327a" animation_style="slide" animation_direction="left" animation_intensity_slide="25%" header_font_size_tablet="" header_font_size_phone="30px" header_font_size_last_edited="on|desktop" body_font_size_tablet="" body_font_size_phone="" body_font_size_last_edited="on|desktop" z_index_tablet="500" border_radii="on|25px|25px|25px|25px" global_colors_info="{}" button_bg_color__hover_enabled="on" \ubc84\ud2bc_bg_color__hover="#8fd2ed" \ubc84\ud2bc_\ud14c\ub450\ub9ac_\uc0c9\uc0c1__hover_\ud65c\uc131\ud654="\ucf1c\uc9d0"]<\/p>\n

    \uc6b0\ub9ac\ub294 \ud568\uaed8<\/h2>\n

    \ud560 \uac83\uc774\ub2e4<\/em><\/h2>\n

    \uce58\ub8cc\ubc95\uc744 \ucc3e\uc544\ub77c!<\/h2>\n

    [\/et_pb_cta][\/et_pb_column][et_pb_column type="1_2" _builder_version="4.16" custom_padding="|||" global_colors_info="{}" custom_padding__hover="|||"][et_pb_image src="https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2024\/08\/2024-strip-footer-strip-copy.png" title_text="2024 \uc2a4\ud2b8\ub9bd \ud478\ud130 \uc2a4\ud2b8\ub9bd \ubcf5\uc0ac" _builder_version="4.27.0" _module_preset="\uae30\ubcf8\uac12" custom_margin="35px||||false|false" global_colors_info="{}"][\/et_pb_image][\/et_pb_column][\/et_pb_row][\/et_pb_section]<\/p>","protected":false},"excerpt":{"rendered":"

    [et_pb_section fb_built="1" \uc804\uccb4 \ub108\ube44="\ucf1c\uc9d0" \ube44\ud65c\uc131\ud654\ub428_\ucf1c\uc9d0="\uaebc\uc9d0|\uaebc\uc9d0|\uaebc\uc9d0" _\ube4c\ub354_\ubc84\uc804="4.16" \ud14c\ub450\ub9ac_\ub108\ube44_\ubc14\ub2e5="55px" \ud14c\ub450\ub9ac_\uc0c9\uc0c1_\ubc14\ub2e5="#29327a" \uc7a0\uae40="\uaebc\uc9d0" \uae00\ub85c\ubc8c_\uc0c9\uc0c1_\uc815\ubcf4="{}"][et_pb_\uc804\uccb4 \ub108\ube44_\ud5e4\ub354 _\ube4c\ub354_\ubc84\uc804="4.16" \uc81c\ubaa9_\uae00\uaf34="|||||||||" title_font_size="55" background_color="#29327a" background_image="https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2020\/12\/Aluisio.jpg" background_position="center_left" custom_padding="9vw||9vw||true" custom_padding_tablet="" custom_padding_phone="|56px||" custom_padding_last_edited="on|desktop" title_font_size_tablet="45px" title_font_size_phone="40px" title_font_size_last_edited="on|phone" z_index_tablet="500" custom_css_main_element="background-position: center 18% !important;" global_colors_info="{}"] \ubd88\uba78\ud654\ub41c \uc138\ud3ec \ubc30\uc591 \ud504\ub85c\ud1a0\ucf5c [\/et_pb_fullwidth_header][\/et_pb_section][et_pb_section fb_built="1" use_custom_gutter="on" gutter_width="1" specialty="on" padding_left_1="35px" padding_left_2="35px" padding_2_tablet="|||0px" padding_2_phone="" padding_2_last_edited="on|desktop" module_class_1="sidebar-secondary-nav" module_class="handprint-bg" _builder_version="4.16" background_image="https:\/\/www.progeriaresearch.org\/wp-content\/uploads\/2019\/04\/blue-handprint-only.png" parallax="on" parallax_method="off" \ub0b4\ubd80_\ub108\ube44="100%" \ub0b4\ubd80_\ucd5c\ub300_\ub108\ube44="100%" \uc0ac\uc6a9\uc790 \uc815\uc758_\ud328\ub529="0|0px|54px|0px|false|false" z_index_tablet="500" \ud14c\ub450\ub9ac_\ub108\ube44_\uc704\ucabd="10px" \ud14c\ub450\ub9ac_\uc0c9\uc0c1_\uc704\ucabd="#8fd2ed" [\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=\"Immortalized Cell Culture Protocols\" font_container=\"tag:h1|text_align:center\" use_theme_fonts=\"yes\"][vc_column_text]\r\n

    Immortalized Cell Culture Protocol<\/h2>\r\nWe thank Dr Martin Dorf at Harvard University for generously generating and providing the\u00a0immortalized cell lines to the PRF Cell & Tissue Bank. Thank you.\r\n\r\nPRF cell lines were immortalized using the following protocol:\r\n

    Retrovirus Infection<\/strong><\/h3>\r\nA. Producing Virus:<\/strong> 12-24 hr before transfection, packaging HEK293 cells were plated on a 100-mm plate at 60-80% confluency. Each plate was co-transfected with 12\u03bcg retroviral construct\u00a0pBABE-puro (SV40T) and 12\u03bcg pCL-Ampho Retrovirus Packaging Vector. The culture medium\u00a0was aspirated 8-10 hr after transfection, and replaced with 10ml of complete medium. The\u00a0culture was incubated for an additional 48-72 hr to attain optimal viral titer.\r\n\r\nB. Infecting Target Cells:<\/strong> The target cells were plated on a 100 mm plate 12-18 hr before\u00a0infection. The medium from packaging cells was collected and filtered through a 0.45-\u03bcm\u00a0cellulose acetate or polysulfonic filter. Once the cells were 40-60% confluent, 8 ml virus-containing medium with a final concentration of 10 \u03bcg\/ml polybrene was added. Three rounds\u00a0of infection were performed sequentially, ~12 hrs apart. The medium was replaced after 24 hr\u00a0incubation.\r\n\r\nC. Selection of Stable Cell Lines:<\/strong> 72 hr after infection, the cells were trypsinized, split at 1:3 and\u00a0transferred to two 100 mm plates in the presence of 3 \u03bcg\/ml puromycin, plus one control plate\u00a0without antibiotic. The culture medium was changed every 2 or 3 days for approximately 2 to 4\u00a0weeks to produce stable cell lines.\r\n

    Protocol for subculturing and freezing immortalized fibroblasts<\/b>\u00a0<\/b><\/h2>\r\n\r\n\r\n\r\n\r\n\r\n\r\n\r\n\r\n
    Media<\/strong><\/td>\r\n\u00a0 Trypsin<\/strong><\/td>\r\n<\/tr>\r\n
    DMEM- Invitrogen #11960-044 (high glucose without L-glutamine)<\/td>\r\n\u00a0 Trypsin EDTA C .25% (Invitrogen#25200-056)<\/td>\r\n<\/tr>\r\n
    15% FBS Fetal Bovine Serum<\/td>\r\n\u00a0 Hank\u2019s Balanced Salt Solution<\/b><\/td>\r\n<\/tr>\r\n
    1% (1x) Penicillin-Streptomycin (Invitrogen#15140-122)<\/td>\r\n\u00a0 HBSS- (Invitrogen#14170 (1X)\u00a0 (-) calcium Chloride,\u00a0 (-)magnesium chloride,\u00a0 (-) magnesium sulfate)<\/td>\r\n<\/tr>\r\n
    1% (1X) L-glutamine (Invitrogen#25030-081)<\/td>\r\n\u00a0 DMSO for Freezing<\/b><\/td>\r\n<\/tr>\r\n
    0.75 \u03bcg\/ml puromycin (InvivoGen #ant-pr-1)<\/td>\r\n\u00a0 Cell Culture Grade DMSO<\/td>\r\n<\/tr>\r\n<\/tbody>\r\n<\/table>\r\nCells will arrive frozen on dry ice at a concentration of approximately 5 x 105<\/sup>cells\/vial<\/b>\r\n

    Protocol for thawing fibroblasts<\/b><\/h3>\r\n
      \r\n \t
    1. Thaw cells rapidly in a 37\u00b0C 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 without puromycin.<\/li>\r\n \t
    4. Place flask in 37\u00b0C 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

      Subculturing PRF cell lines<\/b><\/h3>\r\n1) When cells are established and growing, begin to use media containing 0.75 \u03bcg\/ml puromycin. Continue to maintain the cells in media containing 0.75 \u03bcg\/ml puromycin.\r\n\r\n2) Cells should be split when confluent.\r\n\r\n3) To split cells (volumes given are assuming cells are in a T25):\r\n\r\nA. Using sterile technique, remove media and rinse flask with 2-3 ml of sterile HBSS. Remove and discard HBSS.\r\n\r\nB. 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. If cells are not detached after 3 minutes, you may incubate another 1-2 minutes\r\n\r\nC. 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\n\r\nD. 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 in to 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\n\r\nE. Using sterile technique, remove an aliquot for counting on a hemacytometer.\r\n\r\nF. While you are counting cells, the rest of the solution should be spun in the clinical centrifuge for 5 minutes at 1000 rpms.\r\n\r\n4) After calculating your cell count, plate cells at 2.5 x\u00a0105<\/sup>\u00a0cells per T 25 filter top flask.\r\n\r\n5) Cells should be fed every 2-3 days with fresh growth medium containing 0.75 \u03bcg\/ml puromycin.\r\n\r\nFreezing:\r\n<\/b>Cells should be frozen at no less than 5x 105<\/sup> cells \/ml\/cryovial\u00a0in growth media<\/span> containing 10% DMSO and 30% FBS without puromycin<\/strong> and subsequently placed in an isopropanol freezing chamber at -80\u00b0C overnight. Transfer to the liquid nitrogen the next day.\r\n

      For Further Information Please Contact:<\/b><\/h4>\r\nLeslie B. Gordon, MD, PhD\r\nProfessor of Pediatrics Research\r\nWarren Alpert Medical School of Brown University and Department of Pediatrics, Hasbro Children's Hospital, Providence, RI\r\nDepartment of Anesthesia, Children's Hospital Boston and Harvard Medical School, Boston, MA\r\nMedical Director, The Progeria Research Foundation\r\nPhone: 978-535-2594\r\nFax: 508-543-0377\r\nLeslie_Gordon@brown.edu\u00a0<\/a>\r\n<\/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-1008","page","type-page","status-publish","hentry"],"yoast_head":"\nImmortalized | The Progeria Research Foundation<\/title>\n<meta name=\"description\" content=\"Dr Martin Dorf at Harvard University has generously generated and provided the immortalized cell lines to the PRF Cell & Tissue Bank.\" \/>\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\/ko\/immortalized-cell-culture-protocols\/\" \/>\n<meta property=\"og:locale\" content=\"ko_KR\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Immortalized | The Progeria Research Foundation\" \/>\n<meta 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