New non-viral method for gene transfer into primary cells

Authors:
Gresch O, Engel FB, Nesic D, Tran TT, England HM, Hickman ES, Korner I, Gan L, Chen S, Castro-Obregon S, Hammermann R, Wolf J, Muller-Hartmann H, Nix M, Siebenkotten G, Kraus G and Lun K
In:
Source: Methods
Publication Date: (2004)
Issue: 33(2): 151-163
Research Area:
Cancer Research/Cell Biology
Cardiovascular
Dermatology/Tissue Engineering
Immunotherapy / Hematology
Neurobiology
Cells used in publication:
C6
Species: rat
Tissue Origin: brain
CHO-K1
Species: hamster
Tissue Origin: ovarian
293
Species: human
Tissue Origin: kidney
HeLa
Species: human
Tissue Origin: cervix
K-562
Species: human
Tissue Origin: blood
NIH/3T3
Species: mouse
Tissue Origin: embryo
PC-12
Species: rat
Tissue Origin: adrenal
B cell, human
Species: human
Tissue Origin: blood
Neuron, hippo/cortical, rat
Species: rat
Tissue Origin: brain
T cell, human peripheral blood unstim.
Species: human
Tissue Origin: blood
CD34+ cell, human
Species: human
Tissue Origin: blood
Chondrocyte, human (NHAC-kn)
Species: human
Tissue Origin: cartilage
Cardiomyocyte (R-CM), rat
Species: rat
Tissue Origin: heart
B-CLL
Species: human
Tissue Origin: blood
Keratinocyte, (NHEK-neo) human neonatal
Species: human
Tissue Origin: dermal
Platform:
Nucleofectorâ„¢ I/II/2b
Experiment

STAT6 siRNA in primary human lymphocytes. Si-kinase siRNA in primary rat cortical neurons Nucleofection of different vectors into the following primary cells: human B-CLL cells, human CD34+ cells, rat cardiomyocytes, human chondrocytes, porcine chondrocytes, bovine chondrocytes, rat neurons Nucleofection of vectors expressing EGFP into the following primary cells: NHDF-neo, HUVEC, NHDF-adult, human MSC, human keratinocytes-neo, human keratinocytes-adult Nucleofection of vectors expressing H2Kk into the following primary cells: human T cells, human B cells, NHEM-neo, HAoSMC Nucleofection of vectors expressing EGFP into the following cell lines: CHO-K1, HeLa, NIH-3T3, K562, PC-12, C6 Nucleofection of vectors expressing H2Kk into the following cell lines: COS-7, HEK-293

Abstract

The availability of genetically altered cells is an essential prerequisite for many scientific and therapeutic applications including functional genomics, drug development, and gene therapy. Unfortunately, the efficient gene transfer into primary cells is still problematic. In contrast to transfections of most cell lines, which can be successfully performed using a variety of methods, the introduction of foreign DNA into primary cells requires a careful selection of gene transfer techniques. Whereas viral strategies are time consuming and involve safety risks, non-viral methods proved to be inefficient for most primary cell types. The Nucleofector technology is a novel gene transfer technique designed for primary cells and hard-to-transfect cell lines. This non-viral gene transfer method is based on a cell type specific combination of electrical parameters and solutions. In this report, we show efficient transfer of DNA expression vectors and siRNA oligonucleotides into a variety of primary cell types from different species utilizing the Nucleofector technology, including human B-CLL cells, human CD34+ cells, human lymphocytes, rat cardiomyocytes, human, porcine, and bovine chondrocytes, and rat neurons.