Abstract
Lipid formulations for cell transfection are among the most efficient systems for nucleic acid delivery. During the COVID-19 pandemic, lipid-encapsulated RNA (lipid nanoparticles, LNP) has succeeded as a superior vaccine. Moreover, other similar lipid nanocarriers for siRNA are approved and many are on the pipelines. While lipid encapsulation required several devices for the mixing of components, lipoplex technology allows to rapidly mix nucleic acids and positively charged lipids for cell transfection. In vivo, hepatocytes are important target cells of lipid formulated RNAi. This chapter describes the state-of-the-art lipoplex and LPN manufacturing for treating primary hepatocytes with lipid formulations. Furthermore, protocols for isolating murine hepatocytes and for transfecting these cells with pharmaceutically relevant lipid formulations are provided and discussed.
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Acknowledgments
This research was supported by the MINECO Retos PID2020-117941RB-IOO, SAF2016-78711, and SAF2017-87919-R, MINECO PID2019-104878RB-100AEI/10.13039/501100011033, by Comunidad de Madrid (S2017/BMD-3727 EXOHEP-CM and Y2018/NMT-4949 NanoLiver-CM), and co-funded by European Structural and Investment Fund, ERAB Ref. EA 18/14, AMMF 2018/117, UCM-25-2019, the German Research Foundation (SFB/TRR57/P04, SFB 1382-403224013/A02 and DFG NE 2128/2-1), and COST Action CA17112. YAN and FJC are Ramón y Cajal Researcher RYC-2015-17438 and RYC-2014-15242, respectively. FJC is a Gilead Liver Research 2018. The research group belongs to the validated Research Groups Ref. 970935 ¨Liver Pathophysiology¨, 920631 ¨Lymphocyte immunobiology¨, 920361 “Inmunogenética e inmunología de las mucosas” and IBL-6 (imas12-associated).
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Morán, L., Woitok, M.M., Bartneck, M., Cubero, F.J. (2022). Hepatocyte-Directed Delivery of Lipid-Encapsulated Small Interfering RNA. In: Tanimizu, N. (eds) Hepatocytes. Methods in Molecular Biology, vol 2544. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2557-6_6
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DOI: https://doi.org/10.1007/978-1-0716-2557-6_6
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