Transduction of human islets with the lentiviral vector.
Establishment of an efficient gene delivery system for human pancreatic beta cells is important for the development of diabetes-targeted cell therapies. The human immunodeficiency virus type 1 (HIV-1)-derived lentiviral vector is well documented to be an effective gene transfer tool for various types of cells. Thus, we examined the efficiency of lentivirus-mediated gene delivery for human islets. Human islets were isolated using defined protocols for enzymatic dissociation and purification using discontinuous Ficoll gradients with a refrigerated Cobe 2991 machine. Isolated islets were shipped to Japan, cryopreserved for 3 months, and then subjected to transduction. A vesicular stomatitis virus G-protein (VSV-G)-pseudotyped lentiviral vector LtV-NLS/LacZ was produced in 293T cells under the Fugene6 method. 804G extracellular matrices were applied for monolayer formation of islets. Detection of NLS/LacZ expression was performed using X-gal staining. Lentiviral transduction was effective in these monolayer islets.
Kobayashi
N
N
Department of Surgery, Okayama University Medical School, Okayama, Japan. immortal@md.okayama-u.ac.jp
Arata
T
T
Okitsu
T
T
Ikeda
H
H
Kobayashi
K
K
Kosaka
Y
Y
Narushima
M
M
Tanaka
N
N
Lakey
J R T
JR
D015925:Cryopreservation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology / Q000821:virology*; D016086:Lentivirus / Q000235:genetics* / Q000302:isolation & purification; D008562:Membrane Glycoproteins / Q000235:genetics; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins