{"id":10816,"date":"2021-01-28T16:03:57","date_gmt":"2021-01-28T07:03:57","guid":{"rendered":"http:\/\/itsbio.co.kr\/?page_id=10816"},"modified":"2021-01-28T16:04:00","modified_gmt":"2021-01-28T07:04:00","slug":"cb-ipsc","status":"publish","type":"page","link":"http:\/\/itsbio.co.kr\/?page_id=10816","title":{"rendered":"CB-iPSC"},"content":{"rendered":"<p>[et_pb_section fb_built=&#8221;1&#8243; _builder_version=&#8221;4.4.2&#8243; vertical_offset_tablet=&#8221;0&#8243; horizontal_offset_tablet=&#8221;0&#8243; z_index_tablet=&#8221;0&#8243; custom_css_main_element=&#8221;padding:0;||margin:0;&#8221; box_shadow_horizontal_tablet=&#8221;0px&#8221; box_shadow_vertical_tablet=&#8221;0px&#8221; box_shadow_blur_tablet=&#8221;40px&#8221; box_shadow_spread_tablet=&#8221;0px&#8221;][et_pb_row _builder_version=&#8221;4.4.2&#8243; vertical_offset_tablet=&#8221;0&#8243; horizontal_offset_tablet=&#8221;0&#8243; z_index_tablet=&#8221;0&#8243; custom_css_main_element=&#8221;padding-top:0;||padding-bottom:0;||marign-top:0;||marign-bottom:0;&#8221; 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All cell lines are tested to be free of any transgene vector insertion by PCR analysis and thus are authentic footprint-free iPSCs. These lines have all been passaged long-term (up to P30) without differentiation in optimized human iPSC culture media in the presence of inactivated REF or MEF feeder cells. These lines may have greater propensity to differentiate into Hematopoietic Stem Cells (HSCs) or other mesoderm cells due to epigenetic memory. Great cell model systems for any investigative research work including on iPSCs, HSCs, and disease modeling.<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">SKU<\/th>\n<td class=\"col data\" data-th=\"SKU\">T2010<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Species<\/th>\n<td class=\"col data\" data-th=\"Species\">Human (H. sapiens)<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Tissue\/Organ\/Organ System<\/th>\n<td class=\"col data\" data-th=\"Tissue\/Organ\/Organ System\">Blood<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Vector Map<\/th>\n<td class=\"col data\" data-th=\"Vector Map\"><a href=\"https:\/\/www.abmgood.com\/vectormap\/index\/index\/?name=pEV-SFFV-Oct4-2A-Sox2\" target=\"_blank\" rel=\"noopener\">pEV-SFFV-Oct4-2A-Sox2<\/a><\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Caution<\/th>\n<td class=\"col data\" data-th=\"Caution\">For Research Use Only, not for therapeutic or diagnostic purposes.<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Cell Type<\/th>\n<td class=\"col data\" data-th=\"Cell Type\">Ready-to-Use Stem Cells<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Propagation Requirements<\/th>\n<td class=\"col data\" data-th=\"Propagation Requirements\">The basal medium for this cell line is Prigrow X series medium available at\u00a0<strong>abm<\/strong>, Cat. No.\u00a0<a href=\"https:\/\/www.abmgood.com\/prigrow-x-series-medium-for-t2010-cell-line-tm2010.html\">TM2010<\/a>.<br \/>\nCarbon dioxide (CO<sub>2<\/sub>): 5%, Temperature:37.0\u00b0C.<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Unit quantity<\/th>\n<td class=\"col data\" data-th=\"Unit quantity\">Vial<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Passage Number<\/th>\n<td class=\"col data\" data-th=\"Passage Number\">10<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t<div id=\"tab-108212\" class=\"sp-tab__tab-pane \" role=\"tabpanel\">\n\t\t\t\t\t<div class=\"sp-tab-content \"><p><span class=\"abm-document-subtitle\">Supporting Protocol<\/span><br \/>\n<span class=\"abm-document-subtitle\">MSDS<\/span><br \/>\n<span class=\"abm-document-subtitle\">QC<\/span><br \/>\n<span class=\"abm-document-subtitle\">Other<\/span><\/p>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t<div id=\"tab-108213\" class=\"sp-tab__tab-pane \" role=\"tabpanel\">\n\t\t\t\t\t<div class=\"sp-tab-content \"><style>#sp-ea-10822 .spcollapsing { height: 0; overflow: hidden; transition-property: height;transition-duration: 500ms;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single {border: 1px solid #e2e2e2; }#sp-ea-10822.sp-easy-accordion>.sp-ea-single>.ea-header a {color: #444;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single>.sp-collapse>.ea-body {background: #fff; color: #444;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single {background: #eee;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single>.ea-header a .ea-expand-icon.fa { float: left; color: #444;font-size: 16px;}<\/style><div id=\"sp_easy_accordion-1781929002\"><div id=\"sp-ea-10822\" class=\"sp-ea-one sp-easy-accordion\" data-ex-icon=\"fa-minus\" data-col-icon=\"fa-plus\"  data-ea-active=\"ea-click\"  data-ea-mode=\"vertical\" data-preloader=\"\" data-scroll-active-item=\"\" data-offset-to-scroll=\"0\"><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108220 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> How do I culture these cells?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108220\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>Human iPSCs can be cultured with the Human iPSC Culture Medium Kit (Cat.No.: TM020) together with feeder cells such as MITC Inactivated Rat Embryonic Fibroblast (REF) Feeder Cells (T2017) or MITC Treated Immortalized Mouse Embryonic Fibroblast Feeder Cells (T2009).<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108221 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> How do I ensure the survival of first passage?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108221\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>To improve survival, it is recommended to use ROCK inhibitor (10uM) and conditioned medium immediately upon thawing. Conditioned medium can be made by adding Human iPSC Culture Medium (TM020) to inactivated MEF (T2009) or REF (T2017) for one day and harvest the supernatant.<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108222 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> How do I ensure survival of first passage?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108222\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>To improve survival, it is recommended to use ROCK inhibitor (10uM) and conditioned medium upon thawing. Conditioned medium can be prepared by adding the Human iPSC Culture Medium (TM020) to inactivated MEF (T2009) or REF (T2017) for one day and harvest the supernatant.<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108223 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> What is the protocol for thawing and culturing iPSCs?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108223\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>1) One day before thawing or passaging iPSCs, prepare feeders in 0.1% gelatin-precoated 6-well TC plates (37C for ~1 hour). Seed ~3x10^5 feeder cells per well. 2) Thaw CB-iPSCs by immersing the vial in 37C water bath for 1-2 minutes. Gently pipette 2-3 times and transfer all the cells to a 15 or 50-ml tube with 5-10 ml TM020 medium. 3) Spin down at 200g for 3-5 minutes. Carefully aspirate all the supernatent and resuspend cells in 2 ml TM020 medium. Add Rock inhibitor to increase cell survival. Aspirate MEF medium in the wells with feeder and transfer thawed cells to the feeder for culture. 4) Starting the next day, change 2 ml medium every day or every other day without Rock inhibitor. 1-2 days later, you should be able to see survived small colonies. 5) Every 5-7 days, split iPSCs. Aspirate out medium and add 1 ml 0.5 mM EDTA\/PBS. Incubate 3-5 minutes at 37C. When colonies start to curl up, use 1 ml pipettor to gently break down colonies into clumps of 10-50 cells. transfer 10-25% colonies into a new well precoated with feeders. 6) After 1-2 passages on feeder cells, iPSCs can be cultured with Matrigel-precoated 6-wells using TeSR medium.<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108224 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> What is the protocol for freezing iPSCs?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108224\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>1) 4-5 days after cell split, when iPSCs reach a confluency of 30-60%, aspirate out spend medium and add 1ml 0.5mM EDTA\/PBS. Incubate cells at 37C for ~5min. 2) When colonies start to detach, gently harvest all the cells into a 5-ml or 15-ml or 50-ml tube by pippeting 1-2 times. Avoid multiple pippetting. Breaking down clumps into single cells may substantially decrease cell survival. May add iPSC medium to harvest all the cells. 3) Spin down at 200g for 2-3 minutes. 4) Carefully aspirate out supernatent and gently resuspend cells in 0.5ml iPSC medium. 5) Add 0.5ml freezing medium (TM023) and ROCK inhibitor. Mix well by flipping the vial and transfer the medium to a cryovial. 6) Finally transfer the vial to -80C freezer for short-term storage (days to weeks). After cells are frozen, you may then transfer the vial to liquid nitrogen tank for long-term storage (years).<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108225 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> What is the recommended seeding density for iPSC?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108225\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>IPSCs require seeder cells before thawing and plating the IPSC. Seed ~3x10^5 feeder cells in each well of a 6-well plate. After thawing the iPSCs, put all the cells from the vial into 1 well. After several days, you may start splitting the iPSCs into multiple wells depending on how many cells survived. The density mentioned above is for the seeder cells which should be at 3x10^5. Afterwards, please plate the entire IPSC into one well at 10^6 concentration.<\/p>\n<\/div><\/div><\/div><\/div><\/div>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t<div id=\"tab-108214\" class=\"sp-tab__tab-pane \" role=\"tabpanel\">\n\t\t\t\t\t<div class=\"sp-tab-content \"><p>There are no references for this product yet!<\/p>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\n<p>[\/et_pb_text][\/et_pb_column][\/et_pb_row][\/et_pb_section]<\/p>\n","protected":false},"excerpt":{"rendered":"<p>HOME abm Cell Biology Stem Cells and iPSCs CB-iPSC CB-iPSC CAT.NO UNIT T2010 Vial <style id=\"sp_tab_dynamic_style10821\">#sp-wp-tabs-wrapper_10821 .sp-tab__nav {justify-content: start;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom {display: flex;flex-direction: column-reverse;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom > ul {border-top: 1px solid #cccccc;border-bottom: 0;margin-top: 0;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom > ul > li label.sp-tab__active {border-color: transparent #cccccc#cccccc;margin-top: -1px;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom > ul > li label,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom > ul > li a,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom > ul > .sp-tab__nav-item {border-top: 0;border-top-left-radius: 0;border-top-right-radius: 0;border-bottom-left-radius: 2px;border-bottom-right-radius: 2px;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom > ul {border-bottom: none;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default.sp-tab__horizontal-bottom .sp-tab__tab-content .sp-tab__tab-pane {border-top: 1px solid #cccccc;border-bottom: 0;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default .sp-tab__tab-content .sp-tab-content > ul,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default .sp-tab__tab-content .sp-tab-content > ol {border-bottom: none;}#sp-wp-tabs-wrapper_10821 > .sp-tab__nav-tabs .sp-tab__nav-link.sp-tab__active .sp-tab__tab_title,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default label > .sp-tab__card-header {color: #444;}#sp-wp-tabs-wrapper_10821 > .sp-tab__nav-tabs > .sp-tab__nav-item.show .sp-tab__nav-link,#sp-wp-tabs-wrapper_10821 > .sp-tab__nav-tabs > .sp-tab__nav-item .sp-tab__nav-link.sp-tab__active,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default label > .sp-tab__card-header {background-color: #fff;}#sp-wp-tabs-wrapper_10821 .sp-tab__nav-tabs .sp-tab__nav-item.show .sp-tab__nav-link,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default ul li label.sp-tab__active {border-color: #cccccc#cccccc transparent;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul > li > label,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul > li > a {cursor: pointer;border-color: #cccccc;padding-top: 15px;padding-right: 15px;padding-bottom: 15px;padding-left: 15px;}#sp-wp-tabs-wrapper_10821 > .sp-tab__nav-tabs .sp-tab__nav-link {border: 1px solid #cccccc;height: 100%;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul > li label,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul > li a,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul > .sp-tab__nav-item {border-top-left-radius: 2px;border-top-right-radius: 2px;}#sp-wp-tabs-wrapper_10821 .sp-tab__nav-tabs .sp-tab__nav-item {margin-bottom: -1px; }#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul .sp-tab__nav-item,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default label.collapsed > .sp-tab__card-header {background-color: #ffffff;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul .sp-tab__nav-item {margin-right: 0px;margin-top: 5px;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul .sp-tab__nav-item label:hover .sp-tab__tab_title,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul .sp-tab__nav-item a:hover .sp-tab__tab_title,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default label.collapsed .sp-tab__card-header:hover {color: #444;transition: .3s;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul .sp-tab__nav-item:hover,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default label.collapsed > .sp-tab__card-header:hover {background-color: #fff;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul .sp-tab__nav-item:last-child {margin-right: 0;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > .sp-tab__tab-content .sp-tab__tab-pane {border: 1px solid #cccccc;padding-top: 20px;padding-right: 20px;padding-bottom: 20px;padding-left: 20px;border-top: 0px;background-color: #ffffff;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul {border-bottom: 1px solid #cccccc;}@media(max-width:480px) {#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul li.sp-tab__nav-item {width: 100%;margin-right: 0px;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul li.sp-tab__nav-item:last-child {margin-bottom: -1px;}}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > ul .sp-tab__nav-item .sp-tab__tab_title,#sp-wp-tabs-wrapper_10821.sp-tab__lay-default label > .sp-tab__card-header {font-weight: 600;font-style: normal;font-size: 16px;line-height: 22px;letter-spacing: 0px;color: #444;margin: 0px;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > .sp-tab__tab-content .sp-tab__tab-pane {font-weight: 400;font-style: normal;font-size: 16px;line-height: 24px;letter-spacing: 0px;color: #444;}#sp-wp-tabs-wrapper_10821.sp-tab__lay-default > .sp-tab__tab-content .sp-tab__tab-pane ul li a, #sp-wp-tabs-wrapper_10821.sp-tab__lay-default .sp-tab__tab-content .sp-tab__tab-pane ol li a {color: #444;}<\/style>\t\t<div id=\"sp-wp-tabs-wrapper_10821\" class=\"sp-tab__lay-default\" data-preloader=\"1\" data-activemode=\"tabs-activator-event-click\">\n\t\t\t<div class=\"sp-tab__preloader\">\n\t\t<div class=\"sp-tab__spinner\">\n\t\t\t<div class=\"rect1\"><\/div>\n\t\t\t<div class=\"rect2\"><\/div>\n\t\t\t<div class=\"rect3\"><\/div>\n\t\t\t<div class=\"rect4\"><\/div>\n\t\t\t<div class=\"rect5\"><\/div>\n\t\t<\/div>\n\t<\/div>\n\t<ul class=\"sp-tab__nav sp-tab__nav-tabs\" id=\"sp-tab__ul\" role=\"tablist\">\n\t\t\t\t<li class=\"sp-tab__nav-item\" role=\"tab\">\n\t\t\t\t<label class=\"sp-tab__nav-link sp-tab__active\" data-sptoggle=\"tab\" for=\"#tab-108211\" role=\"tab\" aria-controls=tab-108211 aria-selected=true tabindex=0>\n\t\t\t\t\t<span class=\"tab_title_area\"><H4 class=\"sp-tab__tab_title\">Specifications<\/H4><\/span>\n\t\t\t\t<\/label>\n\t\t\t<\/li>\n\t\t\t\t\t\t<li class=\"sp-tab__nav-item\" role=\"tab\">\n\t\t\t\t<label class=\"sp-tab__nav-link\" data-sptoggle=\"tab\" for=\"#tab-108212\" role=\"tab\" aria-controls=tab-108212 aria-selected=true tabindex=0>\n\t\t\t\t\t<span class=\"tab_title_area\"><H4 class=\"sp-tab__tab_title\">Documents<\/H4><\/span>\n\t\t\t\t<\/label>\n\t\t\t<\/li>\n\t\t\t\t\t\t<li class=\"sp-tab__nav-item\" role=\"tab\">\n\t\t\t\t<label class=\"sp-tab__nav-link\" data-sptoggle=\"tab\" for=\"#tab-108213\" role=\"tab\" aria-controls=tab-108213 aria-selected=true tabindex=0>\n\t\t\t\t\t<span class=\"tab_title_area\"><H4 class=\"sp-tab__tab_title\">FAQs<\/H4><\/span>\n\t\t\t\t<\/label>\n\t\t\t<\/li>\n\t\t\t\t\t\t<li class=\"sp-tab__nav-item\" role=\"tab\">\n\t\t\t\t<label class=\"sp-tab__nav-link\" data-sptoggle=\"tab\" for=\"#tab-108214\" role=\"tab\" aria-controls=tab-108214 aria-selected=true tabindex=0>\n\t\t\t\t\t<span class=\"tab_title_area\"><H4 class=\"sp-tab__tab_title\">References<\/H4><\/span>\n\t\t\t\t<\/label>\n\t\t\t<\/li>\n\t\t\t<\/ul>\n<div class=\"sp-tab__tab-content\">\n\t\t\t\t<div id=\"tab-108211\" class=\"sp-tab__tab-pane sp-tab__show sp-tab__active\" role=\"tabpanel\">\n\t\t\t\t\t<div class=\"sp-tab-content \"><table id=\"product-attribute-specs-table\" class=\"data table additional-attributes product-attribute-specs-table\">\n<tbody>\n<tr class=\"configProduct\">\n<th class=\"label\">Description<\/th>\n<td class=\"col data\" data-th=\"Description\">Cord blood iPSCs induced with non-viral EBNA1-based episomal vector pEV-SFFV-OS. All cell lines are tested to be free of any transgene vector insertion by PCR analysis and thus are authentic footprint-free iPSCs. These lines have all been passaged long-term (up to P30) without differentiation in optimized human iPSC culture media in the presence of inactivated REF or MEF feeder cells. These lines may have greater propensity to differentiate into Hematopoietic Stem Cells (HSCs) or other mesoderm cells due to epigenetic memory. Great cell model systems for any investigative research work including on iPSCs, HSCs, and disease modeling.<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">SKU<\/th>\n<td class=\"col data\" data-th=\"SKU\">T2010<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Species<\/th>\n<td class=\"col data\" data-th=\"Species\">Human (H. sapiens)<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Tissue\/Organ\/Organ System<\/th>\n<td class=\"col data\" data-th=\"Tissue\/Organ\/Organ System\">Blood<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Vector Map<\/th>\n<td class=\"col data\" data-th=\"Vector Map\"><a href=\"https:\/\/www.abmgood.com\/vectormap\/index\/index\/?name=pEV-SFFV-Oct4-2A-Sox2\" target=\"_blank\" rel=\"noopener\">pEV-SFFV-Oct4-2A-Sox2<\/a><\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Caution<\/th>\n<td class=\"col data\" data-th=\"Caution\">For Research Use Only, not for therapeutic or diagnostic purposes.<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Cell Type<\/th>\n<td class=\"col data\" data-th=\"Cell Type\">Ready-to-Use Stem Cells<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Propagation Requirements<\/th>\n<td class=\"col data\" data-th=\"Propagation Requirements\">The basal medium for this cell line is Prigrow X series medium available at\u00a0<strong>abm<\/strong>, Cat. No.\u00a0<a href=\"https:\/\/www.abmgood.com\/prigrow-x-series-medium-for-t2010-cell-line-tm2010.html\">TM2010<\/a>.<br \/>\nCarbon dioxide (CO<sub>2<\/sub>): 5%, Temperature:37.0\u00b0C.<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Unit quantity<\/th>\n<td class=\"col data\" data-th=\"Unit quantity\">Vial<\/td>\n<\/tr>\n<tr class=\"configProduct\">\n<th class=\"label\">Passage Number<\/th>\n<td class=\"col data\" data-th=\"Passage Number\">10<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t<div id=\"tab-108212\" class=\"sp-tab__tab-pane \" role=\"tabpanel\">\n\t\t\t\t\t<div class=\"sp-tab-content \"><p><span class=\"abm-document-subtitle\">Supporting Protocol<\/span><br \/>\n<span class=\"abm-document-subtitle\">MSDS<\/span><br \/>\n<span class=\"abm-document-subtitle\">QC<\/span><br \/>\n<span class=\"abm-document-subtitle\">Other<\/span><\/p>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t<div id=\"tab-108213\" class=\"sp-tab__tab-pane \" role=\"tabpanel\">\n\t\t\t\t\t<div class=\"sp-tab-content \"><style>#sp-ea-10822 .spcollapsing { height: 0; overflow: hidden; transition-property: height;transition-duration: 500ms;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single {border: 1px solid #e2e2e2; }#sp-ea-10822.sp-easy-accordion>.sp-ea-single>.ea-header a {color: #444;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single>.sp-collapse>.ea-body {background: #fff; color: #444;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single {background: #eee;}#sp-ea-10822.sp-easy-accordion>.sp-ea-single>.ea-header a .ea-expand-icon.fa { float: left; color: #444;font-size: 16px;}<\/style><div id=\"sp_easy_accordion-1781929002\"><div id=\"sp-ea-10822\" class=\"sp-ea-one sp-easy-accordion\" data-ex-icon=\"fa-minus\" data-col-icon=\"fa-plus\"  data-ea-active=\"ea-click\"  data-ea-mode=\"vertical\" data-preloader=\"\" data-scroll-active-item=\"\" data-offset-to-scroll=\"0\"><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108220 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> How do I culture these cells?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108220\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>Human iPSCs can be cultured with the Human iPSC Culture Medium Kit (Cat.No.: TM020) together with feeder cells such as MITC Inactivated Rat Embryonic Fibroblast (REF) Feeder Cells (T2017) or MITC Treated Immortalized Mouse Embryonic Fibroblast Feeder Cells (T2009).<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108221 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> How do I ensure the survival of first passage?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108221\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>To improve survival, it is recommended to use ROCK inhibitor (10uM) and conditioned medium immediately upon thawing. Conditioned medium can be made by adding Human iPSC Culture Medium (TM020) to inactivated MEF (T2009) or REF (T2017) for one day and harvest the supernatant.<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108222 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> How do I ensure survival of first passage?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108222\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>To improve survival, it is recommended to use ROCK inhibitor (10uM) and conditioned medium upon thawing. Conditioned medium can be prepared by adding the Human iPSC Culture Medium (TM020) to inactivated MEF (T2009) or REF (T2017) for one day and harvest the supernatant.<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108223 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> What is the protocol for thawing and culturing iPSCs?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108223\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>1) One day before thawing or passaging iPSCs, prepare feeders in 0.1% gelatin-precoated 6-well TC plates (37C for ~1 hour). Seed ~3x10^5 feeder cells per well. 2) Thaw CB-iPSCs by immersing the vial in 37C water bath for 1-2 minutes. Gently pipette 2-3 times and transfer all the cells to a 15 or 50-ml tube with 5-10 ml TM020 medium. 3) Spin down at 200g for 3-5 minutes. Carefully aspirate all the supernatent and resuspend cells in 2 ml TM020 medium. Add Rock inhibitor to increase cell survival. Aspirate MEF medium in the wells with feeder and transfer thawed cells to the feeder for culture. 4) Starting the next day, change 2 ml medium every day or every other day without Rock inhibitor. 1-2 days later, you should be able to see survived small colonies. 5) Every 5-7 days, split iPSCs. Aspirate out medium and add 1 ml 0.5 mM EDTA\/PBS. Incubate 3-5 minutes at 37C. When colonies start to curl up, use 1 ml pipettor to gently break down colonies into clumps of 10-50 cells. transfer 10-25% colonies into a new well precoated with feeders. 6) After 1-2 passages on feeder cells, iPSCs can be cultured with Matrigel-precoated 6-wells using TeSR medium.<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108224 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> What is the protocol for freezing iPSCs?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108224\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>1) 4-5 days after cell split, when iPSCs reach a confluency of 30-60%, aspirate out spend medium and add 1ml 0.5mM EDTA\/PBS. Incubate cells at 37C for ~5min. 2) When colonies start to detach, gently harvest all the cells into a 5-ml or 15-ml or 50-ml tube by pippeting 1-2 times. Avoid multiple pippetting. Breaking down clumps into single cells may substantially decrease cell survival. May add iPSC medium to harvest all the cells. 3) Spin down at 200g for 2-3 minutes. 4) Carefully aspirate out supernatent and gently resuspend cells in 0.5ml iPSC medium. 5) Add 0.5ml freezing medium (TM023) and ROCK inhibitor. Mix well by flipping the vial and transfer the medium to a cryovial. 6) Finally transfer the vial to -80C freezer for short-term storage (days to weeks). After cells are frozen, you may then transfer the vial to liquid nitrogen tank for long-term storage (years).<\/p>\n<\/div><\/div><\/div><div class=\"ea-card  sp-ea-single\"><h3 class=\"ea-header\"><a class=\"collapsed\" data-sptoggle=\"spcollapse\" data-sptarget=#collapse108225 href=\"javascript:void(0)\"  aria-expanded=\"false\"><i class=\"ea-expand-icon fa fa-plus\"><\/i> What is the recommended seeding density for iPSC?<\/a><\/h3><div class=\"sp-collapse spcollapse spcollapse\" id=\"collapse108225\" data-parent=#sp-ea-10822><div class=\"ea-body\"><p>IPSCs require seeder cells before thawing and plating the IPSC. Seed ~3x10^5 feeder cells in each well of a 6-well plate. After thawing the iPSCs, put all the cells from the vial into 1 well. After several days, you may start splitting the iPSCs into multiple wells depending on how many cells survived. The density mentioned above is for the seeder cells which should be at 3x10^5. Afterwards, please plate the entire IPSC into one well at 10^6 concentration.<\/p>\n<\/div><\/div><\/div><\/div><\/div>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t<div id=\"tab-108214\" class=\"sp-tab__tab-pane \" role=\"tabpanel\">\n\t\t\t\t\t<div class=\"sp-tab-content \"><p>There are no references for this product yet!<\/p>\n<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t<\/p>\n","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":"","_et_gb_content_width":"","footnotes":""},"_links":{"self":[{"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=\/wp\/v2\/pages\/10816"}],"collection":[{"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=10816"}],"version-history":[{"count":3,"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=\/wp\/v2\/pages\/10816\/revisions"}],"predecessor-version":[{"id":10823,"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=\/wp\/v2\/pages\/10816\/revisions\/10823"}],"wp:attachment":[{"href":"http:\/\/itsbio.co.kr\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=10816"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}