{"id":35,"date":"2021-10-28T08:15:28","date_gmt":"2021-10-28T08:15:28","guid":{"rendered":"https:\/\/celetrix.com\/celetrix\/?page_id=35"},"modified":"2026-10-23T03:46:29","modified_gmt":"2026-10-23T03:46:29","slug":"stem-cell-therapy","status":"publish","type":"page","link":"https:\/\/www.celetrix.com\/zh\/stem-cell-therapy\/","title":{"rendered":"Stem Cell Research"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-page\" data-elementor-id=\"35\" class=\"elementor elementor-35\" data-elementor-post-type=\"page\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-01b6076 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"01b6076\" 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-15be5a8\" data-id=\"15be5a8\" 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-4cd6619 elementor-widget elementor-widget-bdt-breadcrumbs\" data-id=\"4cd6619\" 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-79cc77d elementor-widget elementor-widget-heading\" data-id=\"79cc77d\" 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<h1 class=\"elementor-heading-title elementor-size-default\">High-Efficiency Electroporation of Human iPSCs<\/h1>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-5a9b531 elementor-widget elementor-widget-text-editor\" data-id=\"5a9b531\" 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>Transfect human iPSCs at &gt;80% efficiency with &gt;85% survival, with no feeder cells needed. Peer-reviewed studies show precise CRISPR editing of iPSC disease models with Celetrix.<\/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-51b6da5 elementor-widget elementor-widget-heading\" data-id=\"51b6da5\" 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\">Results in Human iPSCs<\/h2>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-05bcb39 elementor-widget elementor-widget-text-editor\" data-id=\"05bcb39\" 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>Feeder cells are not required during electroporation. After 3 days, cells grow into compact colonies typical of undifferentiated iPSCs.<\/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-8064c32 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"8064c32\" 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-abf3b8b\" data-id=\"abf3b8b\" 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-b500de8 elementor-widget elementor-widget-image\" data-id=\"b500de8\" 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<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img fetchpriority=\"high\" decoding=\"async\" width=\"412\" height=\"200\" src=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-ipsc-gfp-transfection-24h.jpg\" class=\"attachment-full size-full wp-image-1919\" alt=\"Side-by-side brightfield and fluorescence micrographs showing rapid, high-intensity GFP gene expression and excellent cell viability in human iPSCs 24 hours after electroporation with Celetrix electroporators.\" srcset=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-ipsc-gfp-transfection-24h.jpg 412w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-ipsc-gfp-transfection-24h-300x146.jpg 300w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-ipsc-gfp-transfection-24h-18x9.jpg 18w\" sizes=\"(max-width: 412px) 100vw, 412px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">24 h: GFP expression in ACS-1030 iPSCs<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\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-2974a30\" data-id=\"2974a30\" 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-aea082e elementor-widget elementor-widget-image\" data-id=\"aea082e\" 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<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" width=\"414\" height=\"200\" src=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-undifferentiated-ipsc-colony-72h.jpg\" class=\"attachment-full size-full wp-image-1923\" alt=\"Microscopy images demonstrating sustained GFP expression in intact human iPSC colonies with typical undifferentiated morphology 72 hours after Celetrix electroporation.\" srcset=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-undifferentiated-ipsc-colony-72h.jpg 414w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-undifferentiated-ipsc-colony-72h-300x145.jpg 300w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-acs1030-undifferentiated-ipsc-colony-72h-18x9.jpg 18w\" sizes=\"(max-width: 414px) 100vw, 414px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">72 h: undifferentiated iPSC colonies<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\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<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-071ace9 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"071ace9\" 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-d0c88ef\" data-id=\"d0c88ef\" 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-5853115 elementor-widget elementor-widget-image\" data-id=\"5853115\" 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<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" width=\"436\" height=\"200\" src=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-electroporation-cell-survival-flow-cytometry.jpg\" class=\"attachment-full size-full wp-image-1927\" alt=\"Flow cytometry scatter plots showing high cell survival rate of over 85% for human iPSC after electroporation using Celetrix electroporation system.\" srcset=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-electroporation-cell-survival-flow-cytometry.jpg 436w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-electroporation-cell-survival-flow-cytometry-300x138.jpg 300w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-electroporation-cell-survival-flow-cytometry-18x8.jpg 18w\" sizes=\"(max-width: 436px) 100vw, 436px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Cell survival &gt;85%<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\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-06f0a72\" data-id=\"06f0a72\" 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-56a2379 elementor-widget elementor-widget-image\" data-id=\"56a2379\" 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<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" width=\"430\" height=\"200\" src=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-gfp-transfection-efficiency-flow-cytometry.jpg\" class=\"attachment-full size-full wp-image-1931\" alt=\"Flow cytometry dot plots comparing control and transfected human iPSC with over 80 percent GFP transfection efficiency using the Celetrix electroporation system.\" srcset=\"https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-gfp-transfection-efficiency-flow-cytometry.jpg 430w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-gfp-transfection-efficiency-flow-cytometry-300x140.jpg 300w, https:\/\/www.celetrix.com\/wp-content\/uploads\/celetrix-hipsc-gfp-transfection-efficiency-flow-cytometry-18x8.jpg 18w\" sizes=\"(max-width: 430px) 100vw, 430px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">GFP+ cells &gt;80% vs. control<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\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<div class=\"elementor-element elementor-element-e5c8513 elementor-widget elementor-widget-heading\" data-id=\"e5c8513\" 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\">Published iPSC Gene Editing with Celetrix<\/h2>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-540e64e elementor-widget elementor-widget-text-editor\" data-id=\"540e64e\" 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 style=\"margin:0 0 14px;\">Researchers at Johns Hopkins University School of Medicine used Celetrix electroporation to create precisely edited iPSC disease models.<\/p><div style=\"border:1px solid #dee2e6;border-left:4px solid #0078B5;padding:14px 18px;margin-bottom:14px;\"><p style=\"margin:0 0 4px;font-size:14px;color:#666;\">Scientific Reports, 2021<\/p><p style=\"margin:0 0 6px;font-weight:700;\"><a href=\"https:\/\/doi.org\/10.1038\/s41598-021-89312-2\" target=\"_blank\" rel=\"noopener\" style=\"color:#0078B5;\">Use of single guided Cas9 nickase to facilitate precise and efficient genome editing in human iPSCs \u2197<\/a><\/p><p style=\"margin:0 0 8px;font-size:14px;\">Li PP, Margolis RL. <em>Sci Rep<\/em> 11:9865.<\/p><p style=\"margin:0;\">CRISPR knock-in of an expanded CAG repeat (SCA12 disease model) in human iPSCs electroporated with Celetrix. About 15% of selected clones were precisely edited, with no on-target indels or off-target changes.<\/p><\/div><div style=\"border:1px solid #dee2e6;border-left:4px solid #0078B5;padding:14px 18px;margin-bottom:14px;\"><p style=\"margin:0 0 4px;font-size:14px;color:#666;\">Stem Cell Research, 2021<\/p><p style=\"margin:0 0 6px;font-weight:700;\"><a href=\"https:\/\/doi.org\/10.1016\/j.scr.2021.102346\" target=\"_blank\" rel=\"noopener\" style=\"color:#0078B5;\">Generation of a human induced pluripotent stem cell line JHUi003-A with homozygous mutation for spinocerebellar ataxia type 12 using genome editing \u2197<\/a><\/p><p style=\"margin:0 0 8px;font-size:14px;\">Feng H, Li Q, Margolis RL, Li PP. <em>Stem Cell Res<\/em> 53:102346.<\/p><p style=\"margin:0;\">A homozygous SCA12 iPSC line (69 and 72 CAG repeats) generated with Celetrix electroporation. The line kept a normal karyotype, expressed pluripotency markers (Oct4, Sox2, Nanog) and differentiated into all three germ layers.<\/p><\/div>\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-058ad4a elementor-widget elementor-widget-heading\" data-id=\"058ad4a\" 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\">iPSC Knockout Protocol (Cas9 RNP)<\/h2>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-a1befae elementor-widget elementor-widget-text-editor\" data-id=\"a1befae\" 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 style=\"margin:0 0 10px;\">For a 20 \u00b5l Celetrix tube:<\/p><table class=\"cx-scale-table\" style=\"width:100%;border-collapse:collapse;table-layout:fixed;font-family:Montserrat,sans-serif;font-size:16px;\"><tbody><tr><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;width:26%;font-weight:600;\">Cells<\/td><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;\">1\u20132\u00d710<sup>6<\/sup> cells, pelleted<\/td><\/tr><tr><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;width:26%;font-weight:600;\">Cas9 protein<\/td><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;\">5 \u00b5g<\/td><\/tr><tr><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;width:26%;font-weight:600;\">sgRNA<\/td><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;\">50\u2013100 pmol (for multiple sgRNAs, add Cas9 protein last)<\/td><\/tr><tr><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;width:26%;font-weight:600;\">RNP assembly<\/td><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;\">Mix Cas9 and sgRNA in 22 \u00b5l Celetrix electroporation buffer; incubate 10 min at room temperature<\/td><\/tr><tr><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;width:26%;font-weight:600;\">Electroporation<\/td><td style=\"text-align:left;vertical-align:middle;padding:14px 16px;border-bottom:1px solid #dee2e6;overflow-wrap:anywhere;\">Add the RNP mix to the cell pellet, mix well and electroporate with the <a href=\"\/recommend-parameters\/human-ipsc\/\">recommended human iPSC parameters<\/a> (about 25 \u00b5l total)<\/td><\/tr><\/tbody><\/table><div style=\"background:#F5F8FB;border-left:4px solid #0078B5;padding:14px 18px;margin-top:16px;\"><p style=\"margin:0 0 6px;font-weight:700;color:#0078B5;\">Why RNP instead of Cas9 plasmid?<\/p><ul style=\"margin:0;padding-left:18px;\"><li>Typically &gt;80% knockout efficiency with our RNP protocol<\/li><li>Cas9 plasmid is toxic to cells and lowers survival<\/li><li>Less downstream screening and clone validation work<\/li><\/ul><\/div>\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-47389c6 elementor-widget elementor-widget-heading\" data-id=\"47389c6\" 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\">iPSC Electroporation FAQs<\/h2>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-27ab077 elementor-widget elementor-widget-accordion\" data-id=\"27ab077\" data-element_type=\"widget\" data-widget_type=\"accordion.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div class=\"elementor-accordion\">\n\t\t\t\t\t\t\t<div class=\"elementor-accordion-item\">\n\t\t\t\t\t<div id=\"elementor-tab-title-4151\" class=\"elementor-tab-title\" data-tab=\"1\" role=\"button\" aria-controls=\"elementor-tab-content-4151\" aria-expanded=\"false\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon elementor-accordion-icon-left\" aria-hidden=\"true\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-closed\"><svg class=\"e-font-icon-svg e-fas-plus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H272V64c0-17.67-14.33-32-32-32h-32c-17.67 0-32 14.33-32 32v144H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h144v144c0 17.67 14.33 32 32 32h32c17.67 0 32-14.33 32-32V304h144c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-opened\"><svg class=\"e-font-icon-svg e-fas-minus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h384c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t<a class=\"elementor-accordion-title\" tabindex=\"0\">What is the best way to transfect human iPSCs?<\/a>\n\t\t\t\t\t<\/div>\n\t\t\t\t\t<div id=\"elementor-tab-content-4151\" class=\"elementor-tab-content elementor-clearfix\" data-tab=\"1\" role=\"region\" aria-labelledby=\"elementor-tab-title-4151\"><p>Electroporation is the most efficient way to transfect human iPSCs, which are hard to transfect with lipid reagents. With Celetrix electroporation, human iPSCs reach &gt;80% GFP-positive cells with &gt;85% survival, and no feeder cells are needed.<\/p><\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t<div class=\"elementor-accordion-item\">\n\t\t\t\t\t<div id=\"elementor-tab-title-4152\" class=\"elementor-tab-title\" data-tab=\"2\" role=\"button\" aria-controls=\"elementor-tab-content-4152\" aria-expanded=\"false\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon elementor-accordion-icon-left\" aria-hidden=\"true\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-closed\"><svg class=\"e-font-icon-svg e-fas-plus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H272V64c0-17.67-14.33-32-32-32h-32c-17.67 0-32 14.33-32 32v144H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h144v144c0 17.67 14.33 32 32 32h32c17.67 0 32-14.33 32-32V304h144c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-opened\"><svg class=\"e-font-icon-svg e-fas-minus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h384c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t<a class=\"elementor-accordion-title\" tabindex=\"0\">How can I improve iPSC survival after electroporation?<\/a>\n\t\t\t\t\t<\/div>\n\t\t\t\t\t<div id=\"elementor-tab-content-4152\" class=\"elementor-tab-content elementor-clearfix\" data-tab=\"2\" role=\"region\" aria-labelledby=\"elementor-tab-title-4152\"><p>Dissociate iPSCs into single cells, electroporate promptly, move cells into warm medium right after the pulse, and add RevitaCell or a ROCK inhibitor to the recovery medium. Recover cells in antibiotic-free medium, then switch to regular medium with antibiotics at the first passage. With these steps, Celetrix electroporation keeps &gt;85% of iPSCs alive.<\/p><\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t<div class=\"elementor-accordion-item\">\n\t\t\t\t\t<div id=\"elementor-tab-title-4153\" class=\"elementor-tab-title\" data-tab=\"3\" role=\"button\" aria-controls=\"elementor-tab-content-4153\" aria-expanded=\"false\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon elementor-accordion-icon-left\" aria-hidden=\"true\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-closed\"><svg class=\"e-font-icon-svg e-fas-plus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H272V64c0-17.67-14.33-32-32-32h-32c-17.67 0-32 14.33-32 32v144H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h144v144c0 17.67 14.33 32 32 32h32c17.67 0 32-14.33 32-32V304h144c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-opened\"><svg class=\"e-font-icon-svg e-fas-minus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h384c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t<a class=\"elementor-accordion-title\" tabindex=\"0\">Do iPSCs stay pluripotent after electroporation?<\/a>\n\t\t\t\t\t<\/div>\n\t\t\t\t\t<div id=\"elementor-tab-content-4153\" class=\"elementor-tab-content elementor-clearfix\" data-tab=\"3\" role=\"region\" aria-labelledby=\"elementor-tab-title-4153\"><p>Yes. After Celetrix electroporation, iPSCs grow into undifferentiated colonies by 72 h. An iPSC line edited with Celetrix (<a href=\"https:\/\/doi.org\/10.1016\/j.scr.2021.102346\" target=\"_blank\" rel=\"noopener\">Feng et al., <em>Stem Cell Res<\/em> 2021<\/a>) kept a normal karyotype, expressed Oct4, Sox2 and Nanog, and differentiated into all three germ layers.<\/p><\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t<div class=\"elementor-accordion-item\">\n\t\t\t\t\t<div id=\"elementor-tab-title-4154\" class=\"elementor-tab-title\" data-tab=\"4\" role=\"button\" aria-controls=\"elementor-tab-content-4154\" aria-expanded=\"false\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon elementor-accordion-icon-left\" aria-hidden=\"true\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-closed\"><svg class=\"e-font-icon-svg e-fas-plus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H272V64c0-17.67-14.33-32-32-32h-32c-17.67 0-32 14.33-32 32v144H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h144v144c0 17.67 14.33 32 32 32h32c17.67 0 32-14.33 32-32V304h144c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-opened\"><svg class=\"e-font-icon-svg e-fas-minus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h384c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t<a class=\"elementor-accordion-title\" tabindex=\"0\">How do I deliver Cas9 RNP into iPSCs for gene knockout?<\/a>\n\t\t\t\t\t<\/div>\n\t\t\t\t\t<div id=\"elementor-tab-content-4154\" class=\"elementor-tab-content elementor-clearfix\" data-tab=\"4\" role=\"region\" aria-labelledby=\"elementor-tab-title-4154\"><p>Pre-assemble 5 \u00b5g Cas9 protein with 50\u2013100 pmol sgRNA in 22 \u00b5l Celetrix electroporation buffer for 10 min at room temperature. Add the RNP to 1\u20132 million pelleted cells, mix and electroporate in a 20 \u00b5l Celetrix tube.<\/p><\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t<div class=\"elementor-accordion-item\">\n\t\t\t\t\t<div id=\"elementor-tab-title-4155\" class=\"elementor-tab-title\" data-tab=\"5\" role=\"button\" aria-controls=\"elementor-tab-content-4155\" aria-expanded=\"false\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon elementor-accordion-icon-left\" aria-hidden=\"true\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-closed\"><svg class=\"e-font-icon-svg e-fas-plus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H272V64c0-17.67-14.33-32-32-32h-32c-17.67 0-32 14.33-32 32v144H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h144v144c0 17.67 14.33 32 32 32h32c17.67 0 32-14.33 32-32V304h144c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-opened\"><svg class=\"e-font-icon-svg e-fas-minus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h384c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t<a class=\"elementor-accordion-title\" tabindex=\"0\">Can electroporation be used for CRISPR knock-in in iPSCs?<\/a>\n\t\t\t\t\t<\/div>\n\t\t\t\t\t<div id=\"elementor-tab-content-4155\" class=\"elementor-tab-content elementor-clearfix\" data-tab=\"5\" role=\"region\" aria-labelledby=\"elementor-tab-title-4155\"><p>Yes. Researchers co-delivered Cas9 nickase\/sgRNA, donor and BCL-XL plasmids into human iPSCs in a single Celetrix electroporation and obtained precisely edited clones without on-target indels (<em>Scientific Reports<\/em> 2021; <em>Stem Cell Research<\/em> 2021).<\/p><\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t<div class=\"elementor-accordion-item\">\n\t\t\t\t\t<div id=\"elementor-tab-title-4156\" class=\"elementor-tab-title\" data-tab=\"6\" role=\"button\" aria-controls=\"elementor-tab-content-4156\" aria-expanded=\"false\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon elementor-accordion-icon-left\" aria-hidden=\"true\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-closed\"><svg class=\"e-font-icon-svg e-fas-plus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H272V64c0-17.67-14.33-32-32-32h-32c-17.67 0-32 14.33-32 32v144H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h144v144c0 17.67 14.33 32 32 32h32c17.67 0 32-14.33 32-32V304h144c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t<span class=\"elementor-accordion-icon-opened\"><svg class=\"e-font-icon-svg e-fas-minus\" viewBox=\"0 0 448 512\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\"><path d=\"M416 208H32c-17.67 0-32 14.33-32 32v32c0 17.67 14.33 32 32 32h384c17.67 0 32-14.33 32-32v-32c0-17.67-14.33-32-32-32z\"><\/path><\/svg><\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/span>\n\t\t\t\t\t\t\t\t\t\t\t\t<a class=\"elementor-accordion-title\" tabindex=\"0\">How many iPSCs do I need for electroporation?<\/a>\n\t\t\t\t\t<\/div>\n\t\t\t\t\t<div id=\"elementor-tab-content-4156\" class=\"elementor-tab-content elementor-clearfix\" data-tab=\"6\" role=\"region\" aria-labelledby=\"elementor-tab-title-4156\"><p>A 20 \u00b5l Celetrix tube electroporates 1\u20132 million cells, and a 100 \u00b5l tube 5\u201310 million cells. See <a style=\"color: #0078b5;\" href=\"https:\/\/www.celetrix.com\/recommend-parameters\/\">Recommended Parameters<\/a> for settings.<\/p><\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t\t\t<script type=\"application\/ld+json\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@type\":\"FAQPage\",\"mainEntity\":[{\"@type\":\"Question\",\"name\":\"What is the best way to transfect human iPSCs?\",\"acceptedAnswer\":{\"@type\":\"Answer\",\"text\":\"<p>Electroporation is the most efficient way to transfect human iPSCs, which are hard to transfect with lipid reagents. With Celetrix electroporation, human iPSCs reach &gt;80% GFP-positive cells with &gt;85% survival, and no feeder cells are needed.<\\\/p>\"}},{\"@type\":\"Question\",\"name\":\"How can I improve iPSC survival after electroporation?\",\"acceptedAnswer\":{\"@type\":\"Answer\",\"text\":\"<p>Dissociate iPSCs into single cells, electroporate promptly, move cells into warm medium right after the pulse, and add RevitaCell or a ROCK inhibitor to the recovery medium. Recover cells in antibiotic-free medium, then switch to regular medium with antibiotics at the first passage. With these steps, Celetrix electroporation keeps &gt;85% of iPSCs alive.<\\\/p>\"}},{\"@type\":\"Question\",\"name\":\"Do iPSCs stay pluripotent after electroporation?\",\"acceptedAnswer\":{\"@type\":\"Answer\",\"text\":\"<p>Yes. After Celetrix electroporation, iPSCs grow into undifferentiated colonies by 72 h. An iPSC line edited with Celetrix (<a href=\\\"https:\\\/\\\/doi.org\\\/10.1016\\\/j.scr.2021.102346\\\" target=\\\"_blank\\\" rel=\\\"noopener\\\">Feng et al., <em>Stem Cell Res<\\\/em> 2021<\\\/a>) kept a normal karyotype, expressed Oct4, Sox2 and Nanog, and differentiated into all three germ layers.<\\\/p>\"}},{\"@type\":\"Question\",\"name\":\"How do I deliver Cas9 RNP into iPSCs for gene knockout?\",\"acceptedAnswer\":{\"@type\":\"Answer\",\"text\":\"<p>Pre-assemble 5 \\u00b5g Cas9 protein with 50\\u2013100 pmol sgRNA in 22 \\u00b5l Celetrix electroporation buffer for 10 min at room temperature. Add the RNP to 1\\u20132 million pelleted cells, mix and electroporate in a 20 \\u00b5l Celetrix tube.<\\\/p>\"}},{\"@type\":\"Question\",\"name\":\"Can electroporation be used for CRISPR knock-in in iPSCs?\",\"acceptedAnswer\":{\"@type\":\"Answer\",\"text\":\"<p>Yes. Researchers co-delivered Cas9 nickase\\\/sgRNA, donor and BCL-XL plasmids into human iPSCs in a single Celetrix electroporation and obtained precisely edited clones without on-target indels (<em>Scientific Reports<\\\/em> 2021; <em>Stem Cell Research<\\\/em> 2021).<\\\/p>\"}},{\"@type\":\"Question\",\"name\":\"How many iPSCs do I need for electroporation?\",\"acceptedAnswer\":{\"@type\":\"Answer\",\"text\":\"<p>A 20 \\u00b5l Celetrix tube electroporates 1\\u20132 million cells, and a 100 \\u00b5l tube 5\\u201310 million cells. See <a style=\\\"color: #0078b5;\\\" href=\\\"https:\\\/\\\/www.celetrix.com\\\/recommend-parameters\\\/\\\">Recommended Parameters<\\\/a> for settings.<\\\/p>\"}}]}<\/script>\n\t\t\t\t\t<\/div>\n\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-5e8a0ac elementor-align-center elementor-widget elementor-widget-button\" data-id=\"5e8a0ac\" data-element_type=\"widget\" data-widget_type=\"button.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t<div class=\"elementor-button-wrapper\">\n\t\t\t\t\t<a class=\"elementor-button elementor-button-link elementor-size-sm\" href=\"https:\/\/www.celetrix.com\/ordering\/\">\n\t\t\t\t\t\t<span class=\"elementor-button-content-wrapper\">\n\t\t\t\t\t\t\t\t\t<span class=\"elementor-button-text\">Request a Quote<\/span>\n\t\t\t\t\t<\/span>\n\t\t\t\t\t<\/a>\n\t\t\t\t<\/div>\n\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-top-column elementor-element elementor-element-f68dc84\" data-id=\"f68dc84\" 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-f5fe199 elementor-nav-menu__align-start elementor-nav-menu--dropdown-none elementor-widget elementor-widget-nav-menu\" data-id=\"f5fe199\" data-element_type=\"widget\" data-settings=\"{&quot;layout&quot;:&quot;vertical&quot;,&quot;submenu_icon&quot;:{&quot;value&quot;:&quot;&lt;svg aria-hidden=\\&quot;true\\&quot; class=\\&quot;e-font-icon-svg e-fas-caret-down\\&quot; viewBox=\\&quot;0 0 320 512\\&quot; xmlns=\\&quot;http:\\\/\\\/www.w3.org\\\/2000\\\/svg\\&quot;&gt;&lt;path d=\\&quot;M31.3 192h257.3c17.8 0 26.7 21.5 14.1 34.1L174.1 354.8c-7.8 7.8-20.5 7.8-28.3 0L17.2 226.1C4.6 213.5 13.5 192 31.3 192z\\&quot;&gt;&lt;\\\/path&gt;&lt;\\\/svg&gt;&quot;,&quot;library&quot;:&quot;fa-solid&quot;}}\" data-widget_type=\"nav-menu.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t<nav aria-label=\"Menu\" class=\"elementor-nav-menu--main elementor-nav-menu__container elementor-nav-menu--layout-vertical e--pointer-background e--animation-fade\">\n\t\t\t\t<ul id=\"menu-1-f5fe199\" class=\"elementor-nav-menu sm-vertical\"><li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-287\"><a href=\"https:\/\/www.celetrix.com\/zh\/genome-editing\/\" class=\"elementor-item\">Genome Editing<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-285\"><a href=\"https:\/\/www.celetrix.com\/zh\/antibody-production\/\" class=\"elementor-item\">Antibody production<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-288\"><a href=\"https:\/\/www.celetrix.com\/zh\/stem-cell-therapy\/\" class=\"elementor-item\">Stem Cell Research<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-286\"><a href=\"https:\/\/www.celetrix.com\/zh\/car-tcar-nk\/\" class=\"elementor-item\">CAR-T&#038;CAR-NK<\/a><\/li>\n<\/ul>\t\t\t<\/nav>\n\t\t\t\t\t\t<nav class=\"elementor-nav-menu--dropdown elementor-nav-menu__container\" aria-hidden=\"true\">\n\t\t\t\t<ul id=\"menu-2-f5fe199\" class=\"elementor-nav-menu sm-vertical\"><li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-287\"><a href=\"https:\/\/www.celetrix.com\/zh\/genome-editing\/\" class=\"elementor-item\" tabindex=\"-1\">Genome Editing<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-285\"><a href=\"https:\/\/www.celetrix.com\/zh\/antibody-production\/\" class=\"elementor-item\" tabindex=\"-1\">Antibody production<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-288\"><a href=\"https:\/\/www.celetrix.com\/zh\/stem-cell-therapy\/\" class=\"elementor-item\" tabindex=\"-1\">Stem Cell Research<\/a><\/li>\n<li class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-286\"><a href=\"https:\/\/www.celetrix.com\/zh\/car-tcar-nk\/\" class=\"elementor-item\" tabindex=\"-1\">CAR-T&#038;CAR-NK<\/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>\n\t\t","protected":false},"excerpt":{"rendered":"<p>Electroporation by Celetrix Page High-Efficiency Electroporation of Human iPSCs Transfect human iPSCs at &gt;80% efficiency with &gt;85% survival, with no feeder cells needed. Peer-reviewed studies show precise CRISPR editing of iPSC disease models with Celetrix. Results in Human iPSCs Feeder cells are not required during electroporation. 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