TY - JOUR
T1 - Ex vivo gene delivery to hepatocytes
T2 - Techniques, challenges, and underlying mechanisms
AU - Gao, Shan
AU - Seker, Erkin
AU - Casali, Monica
AU - Wang, Fangjing
AU - Bale, Shyam Sundhar
AU - Price, Gavrielle M.
AU - Yarmush, Martin L.
N1 - Funding Information:
We thank Drs. Stelios Andreadis and Feng Zhang for providing us dual fluorescent lentivirses (GAS-RE), pVSV-G and p∆ vectors, respectively. Authors gratefully acknowledge support by NIH/NBIB (P41) EB002503 and Shriners Hospitals for Children in Boston. ES acknowledges support by Massachusetts General Hospital Fund for Medical Discovery award 217035.
PY - 2012/9
Y1 - 2012/9
N2 - Gene delivery to primary hepatocytes is an important tool for a number of applications including the study of liver cell biology and pathology, drug screening, and gene therapy. Robust transfection of primary hepatocytes, however, is significantly more difficult to achieve than in cell lines or readily dividing primary cells. In this report, we investigated in vitro gene delivery to both primary rat hepatocytes and Huh7.5.1 cells (a hepatoma cell line) using a number of viral and non-viral methods, including Lipofectamine 2000, FuGene HD, Nucleofection, Magnetofection, and lentiviruses. Our results showed that Lipofectamine 2000 is the most efficient reagent for green fluorescent protein (GFP) gene delivery to primary rat hepatocytes (33.3 ± 1.8% transfection efficiency) with minimal adverse effect on several hepatic functions, such as urea and albumin secretion. The lentiviral vectors used in this study exhibited undetectable gene delivery to primary rat hepatocytes but significant delivery to Huh7.5.1 cells (>80% transfection efficiency). In addition, we demonstrated lentiviral-based and spatially defined delivery of the GFP gene to Huh7.5.1 cells for use in biological microelectromechanical systems.
AB - Gene delivery to primary hepatocytes is an important tool for a number of applications including the study of liver cell biology and pathology, drug screening, and gene therapy. Robust transfection of primary hepatocytes, however, is significantly more difficult to achieve than in cell lines or readily dividing primary cells. In this report, we investigated in vitro gene delivery to both primary rat hepatocytes and Huh7.5.1 cells (a hepatoma cell line) using a number of viral and non-viral methods, including Lipofectamine 2000, FuGene HD, Nucleofection, Magnetofection, and lentiviruses. Our results showed that Lipofectamine 2000 is the most efficient reagent for green fluorescent protein (GFP) gene delivery to primary rat hepatocytes (33.3 ± 1.8% transfection efficiency) with minimal adverse effect on several hepatic functions, such as urea and albumin secretion. The lentiviral vectors used in this study exhibited undetectable gene delivery to primary rat hepatocytes but significant delivery to Huh7.5.1 cells (>80% transfection efficiency). In addition, we demonstrated lentiviral-based and spatially defined delivery of the GFP gene to Huh7.5.1 cells for use in biological microelectromechanical systems.
KW - Hepatic function
KW - Huh7.5.1
KW - Lentiviruses
KW - Non-viral transfection
KW - Primary rat hepatocytes
KW - Transfection efficiency
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U2 - 10.1007/s10439-012-0555-y
DO - 10.1007/s10439-012-0555-y
M3 - Article
C2 - 22484829
AN - SCOPUS:84865150034
VL - 40
SP - 1851
EP - 1861
JO - Annals of Biomedical Engineering
JF - Annals of Biomedical Engineering
SN - 0090-6964
IS - 9
ER -