{"id":750,"date":"2022-12-13T02:30:26","date_gmt":"2022-12-13T02:30:26","guid":{"rendered":"https:\/\/www.celetrix.com\/?page_id=750"},"modified":"2023-01-17T09:49:00","modified_gmt":"2023-01-17T09:49:00","slug":"human-cultured-t-cells","status":"publish","type":"page","link":"https:\/\/www.celetrix.com\/zh\/recommend-parameters\/human-cultured-t-cells\/","title":{"rendered":"T cell, cultured or stimulated"},"content":{"rendered":"<div data-elementor-type=\"wp-page\" data-elementor-id=\"750\" class=\"elementor elementor-750\" data-elementor-post-type=\"page\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-57c9258 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"57c9258\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-top-column elementor-element elementor-element-392691a\" data-id=\"392691a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-38a0b89 elementor-widget elementor-widget-bdt-breadcrumbs\" data-id=\"38a0b89\" data-element_type=\"widget\" data-widget_type=\"bdt-breadcrumbs.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class='bdt-breadcrumbs-wrapper'><ul class=\"bdt-breadcrumb breadcrumb-align-left\"><li><a href=\"https:\/\/www.celetrix.com\/zh\/\"><span class=\"bdt-breadcrumbs-home-icon\"> <i aria-hidden=\"true\" class=\"fas fa-home\"><\/i> <\/span>Electroporation by Celetrix<\/a><\/li>\n<li><span> \u9875\u9762<\/span><\/li>\n<\/ul><\/div>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-2f4fa3a elementor-widget elementor-widget-heading\" data-id=\"2f4fa3a\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<h2 class=\"elementor-heading-title elementor-size-default\">Human cultured (stimulated) T cells<\/h2>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-9f80412 elementor-widget elementor-widget-text-editor\" data-id=\"9f80412\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t<p>\u00a0 \u00a0 \u00a0 \u00a0When stimulated by CD3 and CD28 antibodies, T cells grow much larger than fresh PBMC cells. Therefore the voltages used for cultured T cell electroporation is much lower than fresh PBMCs.<\/p><p>\u00a0 \u00a0 \u00a0 \u00a0We have noticed that T cells cultured in different labs require different voltages for electroporation. The variation may be caused by different brands of culture mediums or different CD3\/CD28 antibody and IL-2 combination.<\/p><p>\u00a0 \u00a0 \u00a0 \u00a0 To achieve a good transfection efficiency for cultured T cells while maintaining a good cell proliferative potential, it is imperative that the electroporation voltages be tested out.\u00a0<\/p><p>\u00a0 \u00a0 \u00a0 \u00a0 A special notice for using transposon system with T cells: we noticed that a sleeping beauty transposon system works at a much lower efficiency in stimulated T cells than in unstimulated PBMCs. If you are working with a transposon system such as sleeping beauty or piggybac and having trouble with expression in cultured T cells, try your expression system in fresh PBMCs and it might work much better.<\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-b21ab43 elementor-widget elementor-widget-image\" data-id=\"b21ab43\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<img fetchpriority=\"high\" decoding=\"async\" width=\"400\" height=\"136\" src=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell--e1670898910637.png\" class=\"attachment-full size-full wp-image-753\"  srcset=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell--e1670898910637.png 400w, https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell--e1670898910637-300x102.png 300w, https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell--e1670898910637-18x6.png 18w\" sizes=\"(max-width: 400px) 100vw, 400px\" title=\"T cell, cultured or stimulated\u63d2\u56fe\" alt=\"T cell, cultured or stimulated\u63d2\u56fe\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-ed76460 elementor-widget elementor-widget-image\" data-id=\"ed76460\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" width=\"400\" height=\"190\" src=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell-flowchat-e1670899025553.png\" class=\"attachment-full size-full wp-image-757\"  srcset=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell-flowchat-e1670899025553.png 400w, https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell-flowchat-e1670899025553-300x143.png 300w, https:\/\/www.celetrix.com\/wp-content\/uploads\/2022\/12\/\u57f9\u517bT-cell-flowchat-e1670899025553-18x9.png 18w\" sizes=\"(max-width: 400px) 100vw, 400px\" title=\"T cell, cultured or stimulated\u63d2\u56fe1\" alt=\"T cell, cultured or stimulated\u63d2\u56fe1\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-eb6e9fe elementor-widget elementor-widget-text-editor\" data-id=\"eb6e9fe\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t<p><strong>\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0Transfection efficiency with GFP plasmid after 5 days is over 90%<\/strong><\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-d9fdd2f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"d9fdd2f\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-dad0ca8\" data-id=\"dad0ca8\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-5c0162d elementor-widget elementor-widget-text-editor\" data-id=\"5c0162d\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t<h5 style=\"text-align: center;\"><strong>T cell, cultured or stimulated<br \/>in 20ul tubes<\/strong><\/h5><p>\u00a0 Cell Number\uff1a1-2*10^6 cells<\/p><p>\u00a0 Plasmid\uff1a2ug<\/p><p>\u00a0 E buffer\uff1aCeletrix buffer<\/p><p>\u00a0 Suggested parameter\uff1aCell Line Mode\uff0c480V-540V<\/p><p>\u00a0 Culture medium: antiCD3\/antiCD28\/IL2\u00a0\u00a0(antibiotics free)<\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-2610744\" data-id=\"2610744\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-c424b09 elementor-widget elementor-widget-text-editor\" data-id=\"c424b09\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t<h5 style=\"text-align: center;\"><strong>T cell, cultured or stimulated<br>in 100ul tubes<\/strong><\/h5>\n<p>&nbsp; Cell Number\uff1a5-10*10^6 cells<\/p>\n<p>&nbsp; Plasmid\uff1a10ug<\/p>\n<p>&nbsp; E buffer\uff1aCeletrix buffer<\/p>\n<p>&nbsp; Suggested parameter\uff1aCell Line Mode\uff0c480V-540V<\/p>\n<p>&nbsp; Culture medium: antiCD3\/antiCD28\/IL2&nbsp;&nbsp;(antibiotics free)<\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-top-column elementor-element elementor-element-da60676\" data-id=\"da60676\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-678ff1b elementor-nav-menu__align-start elementor-nav-menu--dropdown-none elementor-widget elementor-widget-nav-menu\" data-id=\"678ff1b\" data-element_type=\"widget\" data-settings=\"{&quot;layout&quot;:&quot;vertical&quot;,&quot;submenu_icon&quot;:{&quot;value&quot;:&quot;&lt;i class=\\&quot;fas fa-caret-down\\&quot; 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Videos<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-107\"><a href=\"https:\/\/www.celetrix.com\/zh\/recommend-parameters\/\" class=\"elementor-item\" tabindex=\"-1\">Recommend Parameters<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-106\"><a href=\"https:\/\/www.celetrix.com\/zh\/optimization-strategy\/\" class=\"elementor-item\" tabindex=\"-1\">Optimization Strategy<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-104\"><a href=\"https:\/\/www.celetrix.com\/zh\/experimental-overview\/\" class=\"elementor-item\" tabindex=\"-1\">Experimental Overview<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-108\"><a href=\"https:\/\/www.celetrix.com\/zh\/special-cells\/\" class=\"elementor-item\" tabindex=\"-1\">Special Cells<\/a><\/li>\n<\/ul>\t\t\t<\/nav>\n\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<\/div>","protected":false},"excerpt":{"rendered":"<p>Electroporation by Celetrix Page Human cultured (stimulated) T cells \u00a0 \u00a0 \u00a0 \u00a0When stimulated by CD3 and CD28 antibodies, T cells grow much larger than fresh PBMC cells. Therefore the voltages used for cultured T cell electroporation is much lower than fresh PBMCs. \u00a0 \u00a0 \u00a0 \u00a0We have noticed that T cells cultured in different [&hellip;]<\/p>","protected":false},"author":1,"featured_media":0,"parent":51,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"elementor_header_footer","meta":{"om_disable_all_campaigns":false,"_uf_show_specific_survey":0,"_uf_disable_surveys":false,"footnotes":""},"class_list":["post-750","page","type-page","status-publish","hentry"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages\/750","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/comments?post=750"}],"version-history":[{"count":19,"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages\/750\/revisions"}],"predecessor-version":[{"id":1390,"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages\/750\/revisions\/1390"}],"up":[{"embeddable":true,"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages\/51"}],"wp:attachment":[{"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/media?parent=750"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}