{"id":3373,"date":"2026-09-02T09:07:43","date_gmt":"2026-09-02T08:07:43","guid":{"rendered":"https:\/\/seamlessbio.de\/?page_id=3373"},"modified":"2026-09-02T09:08:26","modified_gmt":"2026-09-02T08:08:26","slug":"simplacex-hek293-serum-free-medium","status":"publish","type":"page","link":"https:\/\/seamlessbio.de\/de\/simplacex-hek293-serum-free-medium\/","title":{"rendered":"SimplaceX"},"content":{"rendered":"<div data-elementor-type=\"wp-page\" data-elementor-id=\"3373\" class=\"elementor elementor-3373\" data-elementor-post-type=\"page\">\n\t\t\t\t<div class=\"elementor-element elementor-element-be6b73b e-con e-atomic-element e-flexbox-base e-5c49b3f\" data-id=\"be6b73b\" data-element_type=\"e-flexbox\" data-e-type=\"e-flexbox\" data-interaction-id=\"be6b73b\">\n    \t\t<div class=\"elementor-element elementor-element-17512a0 elementor-widget elementor-widget-html\" data-id=\"17512a0\" data-element_type=\"widget\" data-e-type=\"widget\" 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.step{grid-template-columns:1fr}.sb-sfm .cta{padding:28px 20px}}\n<\/style>\n\n<div class=\"sb-sfm\">\n\n<div class=\"bc\">\n  <a href=\"https:\/\/seamlessbio.de\/de\/\">Startseite<\/a><span>\/<\/span>\n  <a href=\"https:\/\/seamlessbio.de\/de\/produkte\/\">Produkte<\/a><span>\/<\/span>\n  <a href=\"https:\/\/seamlessbio.de\/de\/zellkultur\/\">Zellkulturmedien<\/a><span>\/<\/span>\n  <a href=\"https:\/\/seamlessbio.de\/de\/serumfreies-medium-hek293\/\">Serumfreies Medium HEK293<\/a><span>\/<\/span>\n  <strong>SimplaceX<\/strong>\n<\/div>\n\n<div class=\"hero\">\n  <span class=\"ey\">Serum-Free \u00b7 Protein-Supplemented \u00b7 HEK293T \u00b7 EU Manufactured<\/span>\n  <h1>SimplaceX \u2014 Serum-Free Medium for HEK293T<\/h1>\n  <p class=\"sub\">Ready-to-use serum-free medium for HEK293T adherent culture \u2014 optimised for AAV production, lentiviral vector manufacturing and recombinant protein expression. No animal serum. No lot-to-lot FBS variability.<\/p>\n  <div class=\"hbtn\">\n    <a class=\"btn\" href=\"https:\/\/seamlessbio.de\/de\/kontakt\/\">Request Sample or Quote<\/a>\n    <a class=\"btn-o\" href=\"https:\/\/seamlessbio.de\/de\/serumfreies-medium-hek293\/\">All HEK293 Media<\/a>\n  <\/div>\n<\/div>\n\n<div class=\"strip\">\n  <div class=\"cs\"><b>Serumfrei<\/b><span>No FBS \u2014 no lot qualification, no TSE\/BSE documentation burden<\/span><\/div>\n  <div class=\"cs\"><b>AAV-Ready<\/b><span>Optimised for HEK293T triple transfection for AAV and lentiviral vector production<\/span><\/div>\n  <div class=\"cs\"><b>500 mL \/ 2 L<\/b><span>Available in research-scale formats \u2014 larger volumes on request<\/span><\/div>\n  <div class=\"cs\"><b>Data Sheet<\/b><span>Full technical data sheet included with every order<\/span><\/div>\n<\/div>\n\n<h2>What is SimplaceX?<\/h2>\n<p class=\"lede\">SimplaceX is a serum-free, protein-supplemented cell culture medium formulated for HEK293T adherent culture. It replaces FBS as the sole supplement \u2014 providing the growth factors, carrier proteins and lipids required for HEK293T proliferation and transfection efficiency, without animal serum and without the lot-to-lot variability that FBS introduces into AAV and lentiviral vector production.<\/p>\n\n<p>SimplaceX is designed as the first medium for HEK293T serum-free adaptation \u2014 the protein supplement package supports cell attachment and growth during the transition from FBS-containing medium, making it significantly easier to adapt HEK293T cells compared to fully chemically defined media. Once cells are fully adapted to SimplaceX, transition to SimplaceCDX (fully chemically defined) is straightforward if required for GMP or regulatory reasons.<\/p>\n\n<h2>Key Features<\/h2>\n<div class=\"grid\">\n  <div class=\"card\">\n    <h3>No FBS \u2014 no lot qualification<\/h3>\n    <p>Eliminate the FBS lot testing burden from your AAV production process. SimplaceX has a defined protein supplement package \u2014 no lot-to-lot variability in growth factor composition.<\/p>\n  <\/div>\n  <div class=\"card\">\n    <h3>Transfection-compatible<\/h3>\n    <p>Optimised for PEI-mediated triple transfection \u2014 the standard for HEK293T AAV production. Compatible with linear PEI 25 kDa, PEI-Max, and Ca-phosphate.<\/p>\n  <\/div>\n  <div class=\"card\">\n    <h3>Suspension cell rescue protocol<\/h3>\n    <p>HEK293T cells that detach during serum-free adaptation should be rescued, not discarded. SimplaceX is specifically formulated to support these cells when returned to the adherent culture.<\/p>\n  <\/div>\n<\/div>\n\n<h2>Produktinformationen<\/h2>\n<div class=\"tw\">\n  <table>\n    <thead><tr><th>Parameter<\/th><th>Spezifikation<\/th><\/tr><\/thead>\n    <tbody>\n      <tr><td>Format<\/td><td>Serum-free, protein-supplemented \u2014 ready-to-use liquid<\/td><\/tr>\n      <tr><td>Cell type<\/td><td>HEK293T (adherent) \u2014 also suitable for HEK293, HEK293FT<\/td><\/tr>\n      <tr><td>Hauptanwendung<\/td><td>AAV production, lentiviral vector manufacturing, recombinant protein expression<\/td><\/tr>\n      <tr><td>Volumes available<\/td><td>500 mL, 2,000 mL \u2014 larger volumes on request<\/td><\/tr>\n      <tr><td>Frei von tierischen Bestandteilen<\/td><td>No animal serum \u2014 protein supplement from defined sources<\/td><\/tr>\n      <tr><td>Transfektionsvertr\u00e4glichkeit<\/td><td>PEI (linear 25 kDa), PEI-Max, Ca-phosphate, Lipofectamine<\/td><\/tr>\n      <tr><td>pH<\/td><td>7.2\u20137.4<\/td><\/tr>\n      <tr><td>Osmolalit\u00e4t<\/td><td>290\u2013320 mOsm\/kg<\/td><\/tr>\n      <tr><td>Sterilit\u00e4t<\/td><td>0.2 \u00b5m sterile filtered \u2014 sterility tested per lot<\/td><\/tr>\n      <tr><td>Lagerung<\/td><td>2\u20138\u00b0C, protected from light \u2014 stable 12 months from manufacture<\/td><\/tr>\n      <tr><td>Dokumentation<\/td><td>Certificate of Analysis, data sheet \u2014 full regulatory documentation on request<\/td><\/tr>\n      <tr><td>GMP-Signalweg<\/td><td>In development \u2014 contact us for timeline<\/td><\/tr>\n    <\/tbody>\n  <\/table>\n<\/div>\n\n<h2>SimplaceX vs. FBS-Containing Medium<\/h2>\n<div class=\"tw\">\n  <table>\n    <thead><tr><th>Parameter<\/th><th>SimplaceX<\/th><th>DMEM + 10% FBS<\/th><\/tr><\/thead>\n    <tbody>\n      <tr><td>Chargen-zu-Chargen-Schwankungen<\/td><td>\u2705 Defined composition \u2014 no lot qualification needed<\/td><td>\u26a0\ufe0f FBS lot-dependent \u2014 qualification required per lot<\/td><\/tr>\n      <tr><td>Endotoxin<\/td><td>\u2705 &lt;1 EU\/mL \u2014 controlled<\/td><td>\u26a0\ufe0f Variable \u2014 up to 10 EU\/mL in standard FBS<\/td><\/tr>\n      <tr><td>TSE\/BSE-Dokumentation<\/td><td>\u2705 Not required<\/td><td>\u26a0\ufe0f Required \u2014 country-of-origin certification per lot<\/td><\/tr>\n      <tr><td>Nachgeschaltete Reinigung<\/td><td>\u2705 No bovine serum proteins to remove<\/td><td>\u26a0\ufe0f Bovine albumin co-purifies with AAV<\/td><\/tr>\n      <tr><td>GMP-Signalweg<\/td><td>\u2705 Simpler \u2014 animal-free raw material<\/td><td>\u26a0\ufe0f Complex \u2014 FBS highest-risk raw material category<\/td><\/tr>\n      <tr><td>Adaptation required<\/td><td>\u26a0\ufe0f 5\u201310 passages for full adaptation<\/td><td>\u2705 None<\/td><\/tr>\n      <tr><td>AAV titre (optimised)<\/td><td>\u2705 Equivalent to FBS after full adaptation<\/td><td>\u2705 Reference<\/td><\/tr>\n    <\/tbody>\n  <\/table>\n<\/div>\n\n<h2>HEK293T Adaptation Protocol \u2014 SimplaceX<\/h2>\n<p>HEK293T cells adapt to SimplaceX in 5\u201310 passages using a stepwise FBS reduction. The most important principle: <strong>never discard suspension cells during adaptation<\/strong> \u2014 centrifuge them and return them to the flask.<\/p>\n\n<div class=\"step\">\n  <div class=\"step-n\">1<\/div>\n  <div>\n    <h3>Passages 1\u20133 \u2014 Gradual FBS reduction<\/h3>\n    <p>P1: DMEM + 1% FBS + 1\u00d7 SimplaceX &nbsp;|&nbsp; P2\u20133: DMEM + 0.5% FBS + 1\u00d7 SimplaceX<\/p>\n    <p>Low FBS slows the transition deliberately \u2014 more stable adaptation, less cell stress.<\/p>\n  <\/div>\n<\/div>\n\n<div class=\"step\">\n  <div class=\"step-n\">2<\/div>\n  <div>\n    <h3>Passages 4\u20136 \u2014 Fully serum-free SimplaceX<\/h3>\n    <p>DMEM + 1\u00d7 SimplaceX. At every medium change: collect medium \u2192 centrifuge 300 \u00d7 g, 5 min \u2192 resuspend pellet in fresh SimplaceX \u2192 return to flask with trypsinised adherent cells.<\/p>\n  <\/div>\n<\/div>\n\n<div class=\"step\">\n  <div class=\"step-n\">3<\/div>\n  <div>\n    <h3>Detachment \u2014 serum-free protocol<\/h3>\n    <p><strong>Preferred:<\/strong> PBS + 5 mM EDTA \u2014 3\u20135 min at 37\u00b0C. No enzyme inactivation needed.<\/p>\n    <p><strong>Alternative:<\/strong> 0.05% Trypsin-EDTA \u2014 centrifuge immediately to pellet, remove all trypsin, resuspend in SimplaceX.<\/p>\n  <\/div>\n<\/div>\n\n<div class=\"step\">\n  <div class=\"step-n\">4<\/div>\n  <div>\n    <h3>Passages 7\u201325 \u2014 Growth rate recovery<\/h3>\n    <p>Doubling time recovers over 5\u201320 passages. Expect 28\u201342h during early serum-free phase, recovering to 18\u201324h (FBS equivalent) by passage 15\u201325. Continue suspension rescue protocol throughout.<\/p>\n  <\/div>\n<\/div>\n\n<div class=\"note\">\n  <b>Full adaptation protocol:<\/b> The complete step-by-step HEK293T serum-free adaptation protocol including troubleshooting is available as a separate guide \u2014 <a href=\"https:\/\/seamlessbio.de\/de\/ressourcen\/blog\/hek293t-serum-free-adaptation-simplacex\/\">download here<\/a>.\n<\/div>\n\n<h2>Anwendungen<\/h2>\n<div class=\"grid\">\n  <div class=\"card\">\n    <h3>AAV-Produktion<\/h3>\n    <p>Triple transient transfection in HEK293T adherent culture \u2014 AAV2, AAV5, AAV8, AAV9 and novel serotypes. No FBS lot qualification, no BVDV testing burden.<\/p>\n  <\/div>\n  <div class=\"card\">\n    <h3>Lentivirale Vektoren<\/h3>\n    <p>HEK293T triple plasmid transfection for 3rd and 4th generation lentiviral packaging systems \u2014 CAR-T, gene correction, base editing vector production.<\/p>\n  <\/div>\n  <div class=\"card\">\n    <h3>Recombinant Protein<\/h3>\n    <p>Transient or stable recombinant protein expression in HEK293T \u2014 downstream purification simplified by absence of bovine serum proteins.<\/p>\n  <\/div>\n<\/div>\n\n<div class=\"faq\">\n  <h2>H\u00e4ufig gestellte Fragen<\/h2>\n\n  <p class=\"fq\">How many passages does HEK293T adaptation to SimplaceX take?<\/p>\n  <p>Full adaptation typically takes 5\u201310 passages. Growth rate (doubling time) returns to FBS-equivalent levels over 5\u201320 further passages after initial adaptation. The suspension cell rescue protocol at every passage is the most important factor in reducing adaptation time.<\/p>\n\n  <p class=\"fq\">What is the expected AAV titre in SimplaceX compared to FBS?<\/p>\n  <p>After full adaptation, AAV titres in SimplaceX are within 1.5\u00d7 of optimised FBS (Low Endotoxin) in most HEK293T systems. Run a small-scale production test (one 15 cm dish) before committing to full-scale serum-free production.<\/p>\n\n  <p class=\"fq\">Can I use SimplaceX directly without FBS reduction steps?<\/p>\n  <p>Not recommended. A stepwise FBS reduction (1% \u2192 0.5% \u2192 0%) over 3 passages before switching fully to SimplaceX gives significantly better adaptation outcomes than a direct switch. Cells adapt more stably and recover growth rate faster with the gradual approach.<\/p>\n\n  <p class=\"fq\">What is the difference between SimplaceX and SimplaceCDX?<\/p>\n  <p>SimplaceX contains a defined protein supplement package \u2014 it is serum-free but not fully chemically defined. SimplaceCDX is fully chemically defined with no animal-derived or recombinant protein components \u2014 required for processes where CDM documentation is needed for GMP or regulatory filings. Adapt to SimplaceX first, then transition to SimplaceCDX if CDM is required.<\/p>\n\n  <p class=\"fq\">Is SimplaceX available with a data sheet?<\/p>\n  <p>Yes \u2014 a full technical data sheet is included with every order and available on request prior to purchase.<\/p>\n<\/div>\n\n<div class=\"rel\">\n  <h2>\u00c4hnliche Produkte &amp; Anleitungen<\/h2>\n  <div class=\"rg\">\n    <a href=\"https:\/\/seamlessbio.de\/de\/simplacecdx-hek293-chemically-defined-medium\/\">SimplaceCDX \u2014 Chemically Defined \u2192<\/a>\n    <a href=\"https:\/\/seamlessbio.de\/de\/serumfreies-medium-hek293\/\">All HEK293 Serum-Free Media \u2192<\/a>\n    <a href=\"https:\/\/seamlessbio.de\/de\/ressourcen\/blog\/hek293t-serum-free-adaptation-simplacex\/\">Adaptation Protocol Guide \u2192<\/a>\n    <a href=\"https:\/\/seamlessbio.de\/de\/ressourcen\/blog\/serum-free-medium-aav-production-complete-recipe\/\">Serum-Free AAV Medium Components \u2192<\/a>\n    <a href=\"https:\/\/seamlessbio.de\/de\/ressourcen\/blog\/fbs-vs-serum-free-aav-production-when-to-switch\/\">FBS vs Serum-Free \u2014 When to Switch \u2192<\/a>\n    <a href=\"https:\/\/seamlessbio.de\/de\/fbs-mit-geringem-endotoxingehalt\/\">FBS Low Endotoxin (if not yet ready for serum-free) \u2192<\/a>\n  <\/div>\n<\/div>\n\n<div class=\"cta\">\n  <h2>Request SimplaceX \u2014 Sample or Quote<\/h2>\n  <p>Available in 500 mL and 2,000 mL. Full data sheet included. Free test sample on request for adaptation evaluation.<\/p>\n  <div class=\"hbtn\" style=\"justify-content:center\">\n    <a class=\"btn\" href=\"https:\/\/seamlessbio.de\/de\/kontakt\/\">Request Sample or Quote<\/a>\n    <a class=\"btn-o\" href=\"mailto:info@seamlessbio.de?subject=SimplaceX%20Sample%20Request\">info@seamlessbio.de<\/a>\n  <\/div>\n<\/div>\n\n<\/div>\t\t\t\t<\/div>\n\t\t\n<\/div>\n\t\t<\/div>","protected":false},"excerpt":{"rendered":"<p>Home\/ Products\/ Cell Culture Media\/ Serum-Free Media HEK293\/ SimplaceX Serum-Free \u00b7 Protein-Supplemented \u00b7 HEK293T \u00b7 EU Manufactured SimplaceX \u2014 Serum-Free Medium for HEK293T Ready-to-use serum-free medium for HEK293T adherent culture 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