Lipofection of Plasmid DNA into Human Mast Cell Lines using Lipid Nanoparticles Generated by Microfluidic Mixing


Authors: Duguay, B. A., Huang, K. W., & Kulka, M

Journal: Journal of Leukocyte Biology

DOI: 10.1002/JLB.3TA0517-192R

Publication - Abstract

April 18, 2018

Abstract:

Mast cells are important immune cells that have significant roles in mediating allergy and asthma. Therefore, studying the molecular mechanisms regulating these and other processes in mast cells is important to elucidate. Methods such as lipofection, transduction, and electroporation are often employed to dissect these mechanisms by disrupting gene expression in mast cell lines. However, as with other leukocytes, human mast cells (HMCs) are often refractory to the delivery of plasmids by lipofection. In this study, we investigated the utility of lipid nanoparticles (LNPs) containing the ionizable cationic lipids 1,2‐dioleoyloxy‐3‐dimethylaminopropane, 1,2‐dioleyloxy‐3‐dimethylaminopropane, or 2,2‐dilinoleyl‐4‐(2‐dimethylaminoethyl)‐[1,3]‐dioxolane for the delivery of plasmid DNA into HMC lines. Herein, we demonstrate for the first time the use of LNPs to achieve significant and reproducible levels of plasmid DNA transfection in HMC‐1.2 and laboratory of allergic diseases 2 (LAD2) cells. These levels reached 53.2% and 16.0% in HMC‐1.2 and LAD2 cells, respectively; and outperformed Lipofectamine 3000 in both cases. Moreover, cell viability in the transfected cells remained above 65% for all LNP conditions tested. Together, these observations illustrate the efficacy of this technique for mast cell researchers and further support the use of LNPs for nucleic acid delivery into leukocytes.

Advanced Search

close
  • Publications
  • Application Notes
  • Posters
  • Workshops
  • Videos & Webinars
  • Blog Posts
Search

Browse by Category

  • Application
    • Diagnostic and Imaging
    • Genetic Medicine
    • Hematology
    • Metabolic Disorders
    • Neuroscience
    • Oncology
    • Skeletal Disorders
    • Targeted Drug Delivery
    • Vaccines
    • Other Applications
  • Formulation
    • Liposomes
    • Nucleic Acid Lipid Nanoparticles
    • Polymeric Nanoparticles
    • Other Formulations
  • Payload
    • DNA
    • microRNA
    • mRNA
    • siRNA
    • Small Molecule Drugs
    • Other Payloads


related content

Publication - Summary

Effective Lipidoid Nanoparticle Delivery In Vivo of siRNA Targeting Kappa Light Chain Production in a Murine Xenograft Model

Xun Ma, Ping Zhou, Adin Kugelmass, Denis Toskic, Melissa Warner, Lisa X. Lee, Teresa Fogaren, Ming Wang, Yamin Li, Liu Yang, Qiaobing Xu and Raymond Comenzo

Researchers at Tufts University have recently reported a proof of concept for siRNA-LNP treatment for AL amyloidosis - a bone marrow disorder where Immunoglobulin (Ig) light chains (LC) misfold into amyloids that deposit in a variety of other tissues where they cause disfunction ...
Read More


Publication - Summary

An Orthogonal Array Optimization of Lipid-like Nanoparticles for mRNA Delivery in Vivo

Bin Li, Xiao Luo, Binbin Deng, Junfeng Wang, David W. McComb, Yimin Shi, Karin M. L. Gaensler, Xu Tan, Amy L. Dunn, Bryce A. Kerlin, and Yizhou Dong

Although messenger RNA (mRNA) therapeutics are useful for transiently expressing antibodies and proteins, antigen-specific immune responses make the systemic delivery of mRNA challenging.

Read More


Stay Informed

Sign up today to automatically receive new Precision NanoSystems application notes, conference posters, relevant science publications, and webinar invites.

MENU
X