{"id":677,"date":"2022-12-12T02:56:17","date_gmt":"2022-12-12T02:56:17","guid":{"rendered":"https:\/\/www.celetrix.com\/?page_id=677"},"modified":"2023-01-17T09:42:00","modified_gmt":"2023-01-17T09:42:00","slug":"pbmc","status":"publish","type":"page","link":"https:\/\/www.celetrix.com\/zh\/recommend-parameters\/pbmc\/","title":{"rendered":"Human PBMC"},"content":{"rendered":"<div data-elementor-type=\"wp-page\" data-elementor-id=\"677\" class=\"elementor elementor-677\" data-elementor-post-type=\"page\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-ae2641f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ae2641f\" 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-e87960c\" data-id=\"e87960c\" 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-31d195a elementor-widget elementor-widget-bdt-breadcrumbs\" data-id=\"31d195a\" 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-65a2c72 elementor-widget elementor-widget-heading\" data-id=\"65a2c72\" 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 PBMC electroporation\n<\/h2>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-8fcbbec elementor-widget elementor-widget-text-editor\" data-id=\"8fcbbec\" 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 The electroporation conditions for PBMC (peripheral blood mononuclear cells), purified unstimulated T cells or NK cells are similar. Unstimulated T cells have a small cell diameter of only 7-8um and stimulated T cells grow to 10-12um diameter. Because of the small size, PBMC or unstimulated T cells require a much higher electroporaiton voltage than cell lines or stimulated T cells. The voltage is typically 1200V-1400V and only 20ul and 100ul tubes can be used under PBMC mode.<\/p><p>\u00a0 \u00a0 \u00a0PBMCs can also be electroporated in the Legacy mode as we used in the previous Model LE\/EX machines. Under Legacy mode, PBMCs need to be cooled down to 4oC by inserting the electroporation tube completely into ice in an upward direction for 4-6 minutes. After cooling, the cells can be electroporated immediately without wiping off the water drops on the tube. The parameter is 800V-860V 20ms. Model EX+ machine can process 600ul, or up to 200M PBMCs in Legacy mode. The cooling time for 600ul tube is 7-8 minutes and the parameter is 1350-1400V 20ms.<\/p><p>\u00a0 \u00a0 \u00a0Fresh PBMC cells survive quite well after electroporation. The true survival rate is over 60% calculated by the ratio of survived cells\/initial cells. Frozen-thawed PBMC cells need about 30-40V more voltage than fresh PBMC cells.<\/p><p>\u00a0 \u00a0 \u00a0Cell density affects electroporation voltage. For example, increasing from 2M cells to 4M cells in 20ul electroporation will require a voltage increase of about 20V. Donor diference also affects electroporation voltage. There might be +\/- 20V difference for 20ul tubes due to donor difference.\u00a0<\/p><p>\u00a0 \u00a0 \u00a0Transfection efficiency is also heavily influenced by plasmid quality. Good quality GFP plasmid can produce over 80% transfection efficiency. mRNA typically has higher electroporation efficiency and for PBMC, mRNA transfection efficiency is usually &gt;92% with good viability.2<\/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-37be3fd elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"37be3fd\" 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-efd2a2c\" data-id=\"efd2a2c\" 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-9f972de elementor-widget elementor-widget-text-editor\" data-id=\"9f972de\" 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>PBMC in 20ul tubes<\/strong><\/h5><p>\u00a0 Cell Number\uff1a2-10*10^6 cells<\/p><p>\u00a0 Plasmid\uff1a2ug<\/p><p>\u00a0 E buffer\uff1aCeletrix buffer<\/p><p>\u00a0 Suggested parameter\uff1aPBMC Mode\uff0c1200V-1400V<\/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-5441cf1\" data-id=\"5441cf1\" 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-9588bc6 elementor-widget elementor-widget-text-editor\" data-id=\"9588bc6\" 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>PBMC in 100ul tubes<\/strong><\/h5>\n<p>&nbsp; Cell Number\uff1a1-5*10^7 cells<\/p>\n<p>&nbsp; Plasmid\uff1a10ug<\/p>\n<p>&nbsp; E buffer\uff1aCeletrix buffer<\/p>\n<p>&nbsp; Suggested parameter\uff1aPBMC Mode\uff0c1200V-1400V<\/p>\n<p>&nbsp; Culture medium: antiCD3\/antiCD28\/IL2 (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-7e34e47\" data-id=\"7e34e47\" 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-c8a93c4 elementor-nav-menu__align-start elementor-nav-menu--dropdown-none elementor-widget elementor-widget-nav-menu\" data-id=\"c8a93c4\" 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 PBMC electroporation \u00a0 \u00a0 The electroporation conditions for PBMC (peripheral blood mononuclear cells), purified unstimulated T cells or NK cells are similar. Unstimulated T cells have a small cell diameter of only 7-8um and stimulated T cells grow to 10-12um diameter. Because of the small size, PBMC or unstimulated T [&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-677","page","type-page","status-publish","hentry"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages\/677","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=677"}],"version-history":[{"count":30,"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages\/677\/revisions"}],"predecessor-version":[{"id":1386,"href":"https:\/\/www.celetrix.com\/zh\/wp-json\/wp\/v2\/pages\/677\/revisions\/1386"}],"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=677"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}