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- June 24, 2024
- 2:31 pm
Innovations and Applications of Single-Cell Seeding
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Innovations and Applications of Single-Cell Seeding
Advanced single-cell seeding processes continue to revolutionize biomedical research and therapeutic production by ensuring precision and purity. Single-cell seeding minimizes variability and ensures clonality, allowing it to support critical therapeutic areas such as regenerative medicine and oncology. Single-cell seeding accelerates advanced therapy development and high-throughput research, providing a foundation for consistent, high-quality cell populations essential for modern medical advancements. New technologies like the UP.SIGHT from CYTENA eliminates many of the challenges associated with single-cell seeding. In doing so, it allows researchers to streamline the development of novel therapies while remaining compliant with evolving regulatory standards1. Here, we explore the importance of precision in single-cell seeding, its applications, and the pros and cons of different single-cell seeding methods.
Precision in Single-Cell Seeding
Purity and Efficacy
Generating a cell population from a single clone, known as monoclonality, is essential for numerous therapeutic and technological applications2. Precision in single-cell seeding ensures this monoclonality, providing a solid foundation for cell line development (CLD). By ensuring cells are derived from a single, verifiable source, precision in seeding ensures there is no contamination from other clones. This is similar to how quality control checks certify the purity of pharmaceutical drug formulations. This high level of purity is indispensable, as it ensures the therapeutic quality of the cells produced. For instance, monoclonal cell lines ensure consistency, high yield, and efficacy in the final product in biologics production. Precision also reduces variability, which is crucial in applications such as regenerative medicine, where consistent performance is necessary for clinical success.
Compliance and Scalability
Regulatory compliance hinges on traceability, a key advantage of precision single-cell seeding. Regulatory bodies, such as the FDA, scrutinize the purity and origin of potential therapeutics, making traceability essential1. Moreover, precision enables scalability, a crucial factor in modern research and production environments. It allows researchers to streamline workflows, saving on costly reagents and consumables. This scalability is facilitated by using multiwell formats that integrate seamlessly into automated workflows, significantly increasing research throughput. The integration of precision and scalability improves the quality and compliance of the cell lines while boosting productivity and cost-effectiveness in developing advanced therapies.
For further insights, please see our dedicated article on precision in single-cell seeding.
Applications of Single-Cell Seeding in Biomedical Research
Single-cell seeding is revolutionizing biomedical research, offering unprecedented insights into diseases and new treatment options across various fields. Its applications span multiple disease states and drug development phases.
Regenerative Medicine
Single-cell seeding in regenerative medicine allows the isolation and scaling of single stem cells to treat various diseases and conditions. The isolation and manipulation of induced pluripotent stem cells (iPSCs) are crucial for regenerative medicine3. These cells can be differentiated into cell types that replace or grow missing or damaged tissue (Fig. 1). Advances in genetic engineering, particularly CRISPR, allow donor iPSCs to be altered to overcome rejection issues. Accurate single-cell seeding establishes a traceable clonal population, ensuring consistent and replicable differentiation states.
Figure 1. Single-cell dispensing facilitates the isolation and culture of iPSCs, which can be used to regrow a massive variety of tissue types.
Oncology
In oncology, single-cell seeding facilitates the characterization of the myriad of cells within a tumor mass and aids in developing biological treatments like antibodies and cell-based therapies4. It helps us understand treatment resistance, particularly in cancer stem cells (CSCs), which often lead to recurrent, more aggressive tumors5,6. Single-cell seeding also facilitates studying metastasis by allowing circulating tumor cells (CTCs) to be cultured, providing insights into their behavior7.
Therapy Development
For therapy development, single-cell seeding allows drug screening against specific cell populations, allowing researchers to fine-tune dosage, limit toxicity, and enhance efficacy8. It ensures purity and functionality in therapy production, meeting regulatory requirements for traceability. However, single-cell seeding presents biological and technical challenges, such as maintaining optimal growing conditions and achieving accurate dispensing of single cells.
Our dedicated article on single-cell seeding applications dives deeper into these topics.
Methodologies for Single-cell Seeding
Single-cell seeding methods vary across laboratories and are tailored to available resources and equipment. These methods aim to produce clonally distinct populations but differ in efficiency, risk, and consistency9. Accurate seeding is crucial for modern therapeutics, as clonal cell populations are essential for producing cell-based therapies and interventions like antibodies. Regulatory standards require stringent documentation and proof of clonality, which is harder to achieve with some methods than others.
Limiting Dilution and Microfluidics
Limiting dilution involves diluting cells to a low concentration to increase the probability of single-cell dispensing10. However, this method does not guarantee a single cell per well and lacks the efficiency and reproducibility of modern techniques. In contrast, microfluidics technology offers precise control over cellular environments and cell dispensing, making it more efficient and reproducible11. Although initially more expensive, microfluidics systems provide significant advantages over manual methods, helping researchers future-proof their workflows for long-term success.
Automation vs. Manual
Automation in single-cell seeding offers numerous benefits over manual methods, including increased efficiency, reduced human error and contamination risk, and less manual labor. Automation ensures consistent protocols across laboratories, providing long-term savings and scalability. Verifying clonality is essential for regulatory compliance. Manual inspection is time-consuming and error-prone, whereas advanced technologies like CYTENA’s UP.SIGHT’s dual imaging system ensures single-cell dispensing, saving time and providing robust evidence of clonality.
Clone Monitoring and Hit Selection
Advanced imaging methods, such as those used by UP.SIGHT, track cell proliferation and provide valuable insights into cell behavior (Fig. 2). When paired with titer assays for measuring target protein production, like the F.QUANT from CYTENA, advanced imaging streamlines the selection of high-performing clones.
Figure 2. The UP.SIGHT pairs accurate single-cell dispensing with advanced imaging technology to provide >97% single-cell dispensing efficiency and >99.99% probability of clonal derivation.
Our article on basic and advanced methods of single-cell seeding covers these topics in more detail.
Conclusion
Single-cell seeding technologies drive significant advancements in biomedical research and therapeutic production by ensuring precision and minimizing variability. These techniques are fundamental in therapeutic areas like regenerative medicine and oncology, which require verifiable monoclonal cell populations. Technologies like CYTENA’s UP.SIGHT address traditional challenges in single-cell seeding, streamlining the development of novel therapies and making it easy for researchers to demonstrate regulatory compliance. As single-cell seeding evolves, its ability to enhance scalability, traceability, and productivity gives researchers an advantage in the highly competitive cell-based therapy and biologics market.
Ready to speed your novel cell therapy through the discovery phase and beyond? Contact CYTENA’s team of experts today to learn more about the UP.SIGHT all-in-one single-cell dispenser and imager.
References
- World Health Organization. WHO Expert Committee on Biological Standardization. World Health Organ Tech Rep Ser. 2013;(979):1-366, back cover.
- Lai T, Yang Y, Ng SK. Advances in Mammalian cell line development technologies for recombinant protein production. Pharmaceuticals (Basel). 2013;6(5):579-603. doi:10.3390/ph6050579
- Nicholson MW, Ting CY, Chan DZH, et al. Utility of iPSC-Derived Cells for Disease Modeling, Drug Development, and Cell Therapy. Cells. 2022;11(11):1853. doi:10.3390/cells11111853
- Hausser J, Alon U. Tumour heterogeneity and the evolutionary trade-offs of cancer. Nat Rev Cancer. 2020;20(4):247-257. doi:10.1038/s41568-020-0241-6
- Batlle E, Clevers H. Cancer stem cells revisited. Nat Med. 2017;23(10):1124-1134. doi:10.1038/nm.4409
- Damen MPF, Van Rheenen J, Scheele CLGJ. Targeting dormant tumor cells to prevent cancer recurrence. The FEBS Journal. 2021;288(21):6286-6303. doi:10.1111/febs.15626
- Teng T, Yu M. Establishing Single-Cell Clones from In Vitro-Cultured Circulating Tumor Cells. In: Gužvić M, ed. Single Cell Analysis. Vol 2752. Methods in Molecular Biology. Springer US; 2024:119-126. doi:10.1007/978-1-0716-3621-3_8
- Welch JT, Arden NS. Considering “clonality”: A regulatory perspective on the importance of the clonal derivation of mammalian cell banks in biopharmaceutical development. Biologicals. 2019;62:16-21. doi:10.1016/j.biologicals.2019.09.006
- Gross A, Schoendube J, Zimmermann S, Steeb M, Zengerle R, Koltay P. Technologies for Single-Cell Isolation. Int J Mol Sci. 2015;16(8):16897-16919. doi:10.3390/ijms160816897
- Staszewski R. Cloning by limiting dilution: an improved estimate that an interesting culture is monoclonal. Yale J Biol Med. 1984;57(6):865-868.
- Duncombe TA, Tentori AM, Herr AE. Microfluidics: reframing biological enquiry. Nat Rev Mol Cell Biol. 2015;16(9):554-567. doi:10.1038/nrm4041
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