High-throughput transfection and engineering of primary cells and cultured cell lines - an invaluable tool for research as well as drug development

Authors:
Müller-Hartmann H, Faust N, Kazinski M, Kretzschmar T
In:
Source: Exp Opin Drug Discov
Publication Date: (2007)
Issue: 2(11): 1453-1465
Research Area:
Cancer Research/Cell Biology
Cells used in publication:
T cell, human peripheral blood unstim.
Species: human
Tissue Origin: blood
Hepatocyte, mouse
Species: mouse
Tissue Origin: liver
Platform:
Nucleofector™ I/II/2b
96-well Shuttle™ System
Abstract
The manipulation of eukaryotic cells by introducing nucleic acids and other substrates using chemical, physical or viral methods is one of the ground-breaking tools in the life sciences. Changes in the molecular equipment of a cell induced by introducing different molecules not only enable the dissection of signal transduction and metabolic pathways, but also allow the exploitation of engineered cells as bio-factories for the production of proteins in the processes of target research and drug development. In addition to the application of engineered cells for modern cell-based assays, medically relevant engineered cells can be used in clinical settings for adoptive immunotherapy or gene therapy. With the advent of methods exploiting RNA interference (RNAi), gene identification and functional validation in eukaryotic cells have clearly become one of the most exciting methods in life sciences during the past few years. To accelerate research and development in these areas, high-quality, high-throughput approaches (i.e., using sample formats of at least 96 wells) for cell engineering are needed with increasing demand. Recent developments, especially in the field of electroporation, now allow the efficient, high-throughput engineering of virtually any cell type, including primary cells, many of which were previously considered difficult or even impossible to transfect. Primary cells freshly isolated from native tissues are gaining more and more interest, as data obtained with these cells are considered to be of higher physiological relevance than data obtained with immortalized cell lines that have been cultured for extensive periods. In this review, the various methods for cell engineering (with focus on higher eukaryotic cells) are summarized and their impact for high-throughput applications in research and drug development is discussed.