The cell therapy delivery challenge
The possibility of treating disease by modifying a patient’s own cells has captivated researchers and clinicians alike. Cell therapy — including gene-modified approaches such as CAR T and engineered hematopoietic stem cells — is advancing modern medicine, bringing hope to patients with previously untreatable conditions, including cancer, autoimmune diseases, and genetic disorders. The potential of cell-based treatments is immense, from chimeric antigen receptor (CAR) T cell therapies to regenerative medicine. These therapies have shown remarkable success in treating late-stage blood cancers, driving significant growth in the global cell therapy market and fueling a surge in new development programs. But behind every breakthrough is a manufacturing story— one often marked by complexity, cost, and timelines that can stretch over years. Alongside the rapid advancements in cell therapy, we face a significant challenge: the efficient and reliable delivery of therapeutic nucleic acid payloads to target cells in ex vivo, gene-modified cell therapy manufacturing. Traditionally, electroporation (EP) and viral vectors (VV) are the two most widely used methods for gene delivery, each with well-known strengths and drawbacks. EP is often used to introduce messenger RNA (mRNA) or gene-editing tools like CRISPR-Cas9 into primary cells, particularly for transient expression, but it can cause high cell stress and variable outcomes. Among viral delivery platforms, lentiviral and other retroviral vectors are widely used for stable gene integration, while vectors such as adenoviral or adeno-associated viruses are typically applied for transient expression. Regardless of type, viral vector manufacturing often involves long lead times, high costs, and potential safety risks. There can also be short and long-term toxicities associated with persistent expression of the CAR transgene, which has limited access to these therapies.
Using lipid nanoparticles for cell therapy offers a third option. Like EP, LNPs can be used to deliver RNA payloads for the transient expression of a therapeutic gene or gene editing tools. But unlike EP, LNPs are gentler on cells, easier to scale, and simpler to integrate into manufacturing workflows. In some cases, LNPs can also be combined with viral vectors, for example, to achieve persistent expression of a therapeutic gene or targeted gene correction by gene editing. By rethinking delivery and understanding where LNPs complement the existing path or offer a new path for gene delivery, drug developers can unlock new opportunities to design new therapies, scale faster, and reduce costs.
Simplifying cell therapy delivery: Why LNPs are more than an alternative
Lipid nanoparticles represent more than just another tool in gene delivery; they bring a paradigm shift to the field. These nanoparticles are made of biocompatible lipids that encapsulate nucleic acids, enabling targeted delivery into cells through endocytosis. They fuse with the cell membrane and release cargo (Figure 1). Compared to electroporation and viral vectors, LNPs offer numerous advantages in both biological performance and process optimization.
Fig 1. LNP cellular transport pathway. In vivo, lipid nanoparticles typically bind apolipoprotein E (ApoE) in biological fluids, which facilitates interaction with LDL receptors (LDLR) for cellular uptake. While the illustration here shows direct interaction for simplicity, ApoE plays a key intermediary role in this process.
LNPs versus electroporation
- Enhanced cell viability and transfection efficiency: Electroporation often leads to considerable cell damage, resulting in lower yields of viable cells post-transfection. In contrast, LNPs maintain higher cell viability, ensuring more functional cells for downstream applications.
- Simpler cell culture workflow: While EP requires extensive pre- and post-transfection handling (buffer exchange, recovery steps, etc.), LNP delivery can be added directly to cells in one-step, reducing the need for volume reduction and, in many cases, eliminating post-transfection washes, though specific protocols may vary by application and kits.
Jeffrey Lam, Senior Product Manager, Nanomedicine Cell Therapy at Cytiva, emphasizes the operational benefits of using LNP. He shares, “Comparing the workflow between LNPs and electroporation, LNPs are far simpler—fewer washes, fewer manual steps, and the system remains closed. This all translates to lower error rates, contamination risk, batch failures, and high cell recovery.”
LNPs versus viral vectors
- Scalable production: Scaling up viral vector production is intricate and costly. LNPs, however, are chemically defined and have a more predictable scale-up process, making them a viable solution for both small-scale clinical trials and large-scale commercial production.
- Cost efficiency: Viral vectors are expensive to produce, involving complex and time-consuming methods. LNPs eliminate the need for viral production and clearance steps, reducing costs and improving scalability in cell therapy production.
- Closed, modular manufacturing systems: LNP delivery methods are more easily integrated into closed, automated systems, reducing contamination risk and simplifying process integration. This modularity supports flexible manufacturing approaches, from early development to commercial-scale production.
One of the key advantages of LNP delivery is its streamlined cell culture workflow. As Paula Marcus, Manager, Cell Therapy, Biologics and Applications at Cytiva, points out, “Unlike both lentiviral vector and electroporation, LNPs do not require volume reduction, or any additional processes performed on the cells prior to adding the LNPs. No washing or volume increase is required post addition, other than what is normally required for cell division.”
From promise to practice: Real-world applications of LNPs in cell therapy
Cell therapy is evolving, and so are genetic engineering tools. The scientific evidence supporting the use of LNPs in cell therapy is compelling. Several studies and publications highlight the superior performance of LNPs over traditional methods, particularly in terms of flexibility, cell viability, scalability, and process efficiency.
Let’s take a closer look at how LNPs are helping reshape cell therapy delivery through real applications that are already making an impact.
Case study 1 – Rethinking autologous CAR T cells: Precision control
Autologous CAR T therapies engineered using viral vectors for stable expression of CAR have been a game-changer for a small number of blood cancers. However, there remains a strong need to reduce toxicity and enable therapies for new cancer targets. That’s why we are seeing a shift towards RNA-based delivery for transient expression of CAR and other therapeutic proteins.
Researchers at the Fraunhofer IZI conducted a comparative study between LNPs and EP for CAR mRNA delivery (1). LNPs showed improved viability and similar transfection efficiencies compared to EP. Notably, they also demonstrated prolonged CAR T cell efficacy in vitro for LNPs due to extended mRNA persistence and CAR expression. Additionally, CAR expression and in vitro functionality of LNP-modified cells were comparable and less exhausted than lentiviral vector (LVV) transduced cells. The authors also highlight the proven cost- and time-effective manufacturability and scalability of LNPs, outlining a clear path to the clinic.
“LNP delivery enabled efficient CAR expression without the need for electroporation. The process was gentler on cells and led to improved gene expression and cell functionality.”—Dr. Sandy Tretbar, Fraunhofer IZI
Case study 2 – Towards multiplex cell engineering: Smarter CRISPR with LNPs
Gene editing in primary cells can be achieved by different enzymes, with CRISPR-Cas9 increasingly preferred due to ease of target selection and ability to make a range of edits, including knock-outs and knock-ins. The Cas9 protein, encoded from mRNA, is directed to the target DNA by a guide RNA, where it induces double-strand breaks that are mostly resolved through non-homologous end joining (NHEJ), or by homology-directed repair (HDR) in the presence of a DNA template. Many researchers are now trying to combine the benefits of CRISPR-Cas9 and LNP technologies to enable applications ranging from allogeneic T cells therapies to hematopoietic stem cells (HSC) therapies for rare diseases.
Using CRISPR/Cas9 in hematopoietic stem cells has the potential to advance the treatments of genetic blood disorders by correcting disease-causing mutations. Researchers at the SR-Tiget have published a study on developing a novel gene-edited HSC therapy for Hyper IgM syndrome [2]. In this study, they compared LNP and EP to deliver Cas9 mRNA and gRNA targeting CD40L, performed in combination with an adeno-associated virus (AAV) DNA template. RNA nuclease delivery using LNP showed less cell death and improved cell growth compared to EP, yielding a higher number of HDR-edited HSCs, a critical metric for these therapies. The authors use these benefits to discuss the potential of LNPs for concurrent or asynchronous delivery of editing reagents and nucleic acids, endowing cells with specific functions. Overall, this study provides strong evidence that LNPs have the potential to improve the safety and efficiency of ex vivo cell engineering [3].
“LNPs provide added value and versatility for efficient ex vivo hematopoietic cell engineering. Increasing cell recovery over electroporation may improve the efficacy and cost of these therapies by enabling multiplex engineering and shortening the manufacturing process.”—Dr. Samuele Ferrari, SR-Tiget
One of the most ambitious goals in cell therapy is to create a universal cell therapy product that is safe and effective across various types of cancer. An innovative example of this concept comes from researchers at the University of Pennsylvania with a tandem T cell and HSC product utilizing base editing, a derivative of CRISPR-Cas9 designed for making point mutations with reduced risk for chromosomal instability. T cells expressing CAR targeting CD45, a pan-leukocyte marker, and HSCs are separately base-edited with a protective CD45 mutation. This allows the CD45-epitope edited CAR T cells to be protected against fratricide and to re-populate a healthy hematopoietic cell population with the protected HSCs [4]. In follow-up work, the UPenn team is now comparing LNPs to EP for the CD45 base editing. Their results show high efficiency editing and improved cell yields in both T cells and HSCs, with a simplified manufacturing workflow compared to EP (5).
“We aim to base-edit CD45 in HSCs and T cells using LNPs. LNP delivery may provide a more streamlined approach to simplify CAR T cell manufacturing and to increase genetically engineered HSC output during manufacturing.”—Dr. Friederike Herbst, University of Pennsylvania
These case studies show that RNA-based LNPs aren’t just theoretical; they’re already enabling better cell engineering outcomes by giving researchers more ways to tailor therapies and bring them to more patients faster.
- Greater control through transient expression
- Better cell viability and yield compared to EP
- Flexibility to work alongside viral vectors
- Compatibility with GMP-compliant manufacturing
Application-ready kits to simplify and scale LNPs for cell therapy delivery
Application-ready LNP kits are helping simplify the delivery of RNA and gene modification reagents by offering reliable, streamlined solutions that support consistent results from early discovery to clinical production. To meet this need, Cytiva is dedicated to providing innovative tools and solutions to meet the growing demand for cell therapy. Our LNP kits are specially engineered to streamline the cell transfection during preclinical development and provide a path to the clinic.
LNP kits designed to support T cell and CD34+ HSC workflows
The GenVoy-ILM T Cell kits and CD34+ HSC LNP kits are ready-to-use reagents for efficient delivery of mRNA or Cas9 mRNA/sgRNA into primary human T cells and CD34+ hematopoietic stem cells, respectively, offering high gene expression or knockout efficiencies and maintaining viability. While widely used in gene editing workflows, researchers also explore these kits for broader mRNA delivery applications such as immune modulation or reprogramming. The kits support early-stage development with a path to clinic through scalable workflows and support from Cytiva’s services portfolio.
Each of these kits is:
- Intended for research use, with validated performance in key cell types
- Designed for compatibility with Cytiva’s LNP manufacturing and cell therapy manufacturing platforms
- Supported by extensive technical support, training resources, and a clear path to GMP through Cytiva’s services team
These solutions are designed to accelerate the transition from early discovery to clinical trials, streamlining your path from proof of concept to GMP manufacturing.
From discovery to delivery: Cytiva’s end-to-end expertise
Cytiva’s platforms and support services help researchers and manufacturers to seamlessly adopt LNP technology at every stage of the cell therapy development process. Our comprehensive portfolio includes everything you need—from LNP production to cell culture and transfection, backed by a team of professionals who understand the unique challenges of cell therapy manufacturing.
- Nanoparticle formulation: Cytiva’s NanoAssemblr™ instruments and GenVoy-ILM reagents are purpose-built for efficient LNP formulation, ensuring high-quality and scalable LNP production for cell therapy.
- Closed system automation tools: Explore our closed systems and automated cell culture platforms like Sefia™ and Xuri™ systems, designed to support the full lifecycle of cell therapy development from early discovery through clinical production.
- Training and support: We provide hands-on training and guidance throughout the entire process, ensuring that your team has the tools and knowledge to optimize every step of your workflow.
- Regulatory insight: With a deep understanding of GMP and regulatory requirements, Cytiva provides the support needed to bring cell therapies to market with confidence.
Looking ahead: What’s next for LNP-enabled cell therapy
LNPs offer a promising delivery platform for RNA-based gene modification of cell therapies (Advanced therapy medicinal products, or ATMPs), simplifying workflows while supporting GMP manufacturing and quality control. The use of LNPs in cell therapy is in its early stages, but the potential is vast. Ongoing research and innovation are expanding the capabilities of LNP technology, with exciting developments on the horizon:
- DNA delivery for permanent expression: Today’s LNP platforms primarily support RNA delivery for transient expression. But the next critical leap is enabling non-viral DNA delivery ex vivo using LNPs. This could unlock the potential for stable genetic modification, enabling a wide range of new applications and therapies.
- Integrated LNP and automation platforms: Combining LNPs with fully automated cell culture systems like the Sefia™ and Xuri™ platforms will enable standardized workflows, further reduce manual intervention, and increase process efficiency.
At Cytiva, we are proud to be your collaborator on this journey. From discovery to delivery, we provide the tools, technology, and expertise needed to accelerate your progress. Together, let’s bring the promise of cell therapy to life, faster and more efficiently than ever before.
*Author: Reshma Kapatrala, Marketing Writer, Cytiva
Contributors:
- Samuel Clarke, Director, Cytiva
- Jeffrey Lam, Senior Product Manager, Nanomedicine Cell Therapy, Cytiva
- Paula Marcus, Manager, Cell Therapy Biologics and Applications, Cytiva
References
- Reni Kitte, Rabel M, Reka Geczy, et al. Lipid nanoparticles outperform electroporation in mRNA-based CAR T cell engineering. Molecular Therapy - Methods & Clinical Development. 2023;31:101139-101139. doi:https://doi.org/10.1016/j.omtm.2023.101139
- Vavassori V, Ferrari S, Beretta S, et al. Lipid nanoparticles allow efficient and harmless ex vivo gene editing of human hematopoietic cells. Blood. 2023;142(9):812-826. doi:10.1182/blood.2022019333
- Feyisayo Eweje, Bauer DE. A viable alternative for editor delivery. Blood. 2023;142(9):755-756. doi:https://doi.org/10.1182/blood.2023021309
- Wellhausen N, O'Connell RP, Lesch S, et al. Epitope base editing CD45 in hematopoietic cells enables universal blood cancer immune therapy. Sci Transl Med. 2023;15(714):eadi1145. doi:10.1126/scitranslmed.adi1145
- Ghahe EK, Wellhausen N, Geczy R, et al. LIPID NANOPARTICLES ENABLE EFFICIENT BASE EDITING OF CD45-DIRECTED CAR T CELLS FOR UNIVERSAL BLOOD CANCER IMMUNOTHERAPY. Cytotherapy. 2024;26(6):S212. doi:https://doi.org/10.1016/j.jcyt.2024.03.424