iPSCs revolutionized cell therapy by creating patient-specific pluripotent cells, but early viral vector methods carried insertional mutagenesis risks. Non-integrative mRNA-based reprogramming resolves this, delivering transient reprogramming factors for epigenetic remodeling, yielding genomically intact, stable iPSCs critical for clinical translation.
Creative Biolabs' Tailored mRNA-based iPSC Reprogramming Service delivers integration-free, clinical-compliant iPSCs through synthetic mRNA and AI-powered quality control. It mitigates conventional hazards, guarantees batch uniformity, facilitates GMP-scale expansion, and adheres to criteria for regenerative medicine and drug development.
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The key innovation resides in the use of synthetic, modified mRNA for expressing pluripotency factors in vitro—a foundational element of contemporary regenerative medicine.
Fig.1 The generation of iPSC cell lines requires corresponding strategy choices based on clinical conditions and goals, and also involves the participation of mRNA.1,3
Mechanism: Reprogramming involves delivering optimized synthetic mRNA transcripts (often incorporating modifications like pseudouridine to evade the innate immune response) into somatic cells. The mRNA is translated into the key transcription factors (e.g., Oct4, Sox2, Klf4), which initiate the complete epigenetic reset. Once the somatic cell identity is erased and pluripotency is established, the exogenous mRNA naturally degrades, leaving behind a footprint-free iPSC.
Our comprehensive process is structured to provide full transparency and control, guaranteeing a traceable, high-quality final product.
Carry out daily transfections with synthetic, self-amplifying mRNA encoding Yamanaka factors (e.g., OCT4, SOX2, KLF4, GLIS1) to achieve non-integrative reprogramming.
Automatically screen and isolate pluripotent colonies, then expand them under feeder-free conditions. Incorporate an interferon-gamma inhibitor during reprogramming to enhance efficiency, followed by its removal to clear residual mRNA factors.
Adopt a multi-tiered QC approach: karyotype analysis, detection of remaining undifferentiated cells (teratoma risk evaluation), pluripotency marker expression assays, and differentiation potential assessment. Outcome: Fully validated iPSC lines complying with ICH and regulatory criteria for genomic integrity and safety.
Perform large-scale expansion and cryopreservation of fully characterized, clinical-grade iPSC clones in line with GMP-aligned protocols.
Estimated Timeframe: The typical timeframe for this service ranges from 10 to 16 weeks, depending on the source material provided, the desired scale of the Master Cell Bank, and the scope of the final QC panel requested.
Creative Biolabs offers a Custom mRNA based iPS Cell Reprogramming Service that is fully optimized and adjustable to your specific clinical or research requirements, focusing on safety, quality, and industrial readiness.
Our core technology utilizes synthetic, modified mRNA to ensure zero risk of insertional mutagenesis and complete clearance of reprogramming factors.
Customized protocols optimized for various challenging starting materials, including PBMCs, dermal fibroblasts, keratinocytes, and urinary epithelial cells.
Ability to run the reprogramming process with various factor cocktails (OSKM, OSNL, etc.) and small molecule enhancers to maximize efficiency for specific cell types.
Customized post-reprogramming QC panels, including full karyotype analysis, SNP-array analysis, and Whole-Genome Sequencing (WGS), to guarantee chromosomal stability and rule out clonal drift.
Development of a robust MCB using feeder-free, xeno-free media systems, designed for immediate transition to closed-loop bioreactor expansion.
Seamless integration with our downstream services, including CRISPR/Cas9 gene editing (for isogenic correction) and controlled differentiation into target cell lineages.
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Reprogramming was carried out by transfecting BOEC with mixed RNA (including reprogramming factors such as OCT4 and SOX2, immune-escaping RNAs such as E3 and K3, and specific mature double-stranded microRNA clusters) for 8 consecutive days. On the 10th day, clones with iPSC characteristics (closely arranged, clearly demarcated cell clusters) emerged. After karyotype analysis, assessment of undifferentiated state, detection of three-lineage differentiation ability, and STR identification, the karyotype is normal, pluripotent markers such as SOX2 and NANOG are positive, and the blastocyst can differentiate into three germ layers.
Fig.2 BOEC was continuously transfected with an RNA mixture for 8 days, and clones similar to iPSC were selected to detect related markers.2,3
Yes, the service is truly footprint-free. Unlike episomal vectors, which use DNA plasmids that can persist and occasionally integrate into the genome, our synthetic mRNA naturally degrades after the reprogramming factors are expressed. This eliminates the risk of genomic integration, which significantly simplifies the regulatory pathway for clinical applications. Contact us to discuss our validation data.
Absolutely. Our mRNA platform is highly compatible with the introduction of auxiliary gene editing tools. We routinely integrate CRISPR/Cas9-mediated gene correction to create isogenic control lines or correct known patient mutations (e.g., LRRK2 for Parkinson's disease), providing you with the most accurate disease models for therapeutic validation.
Yes, the process is designed for scalability. The stability and genomic integrity attained via mRNA reprogramming render the resulting cell lines ideally suited for closed, large-scale bioreactor expansion — a requirement for GMP production. We deliver the Master Cell Bank with the necessary documentation to integrate directly into your clinical-scale production environment.
Creative Biolabs offers the industry's most advanced, footprint-free solution for the production of clinical-grade iPSCs. By leveraging superior mRNA technology and AI-integrated quality control, we transform the bottlenecks of genomic instability and manufacturing risk into a seamless, confident pathway for your cell therapy program.
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