Detection and Selection of Lentiviral Vector-Transduced Cells

互联网2014-02-13

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Lentiviruses are members of a subgroup of enveloped Retroviridae. Lentivirus-based gene delivery vectors have gained popularity in gene therapy field because of their notable potentials in delivering and integrating transgenes into both mitotic active and inactive cells in vitro and in vivo, a characteristic that overcomes many of the barriers of therapeutic gene therapy application. In the past few years, different lentiviral vector systems have been developed based on primate and nonprimate lentiviruses, including human immunodeficiency virus type 1 (HIV-1), simian immunodeficiency virus (SIV), equine infectious anemia virus (EIAV), and feline immunodeficiency virus (FIV) (see refs. 1 –4 ). However, HIV-1-based lentiviral vector remains the best-studied and well-developed lentiviral vector system that has shown high transduction efficiencies targeting many human and experimental animal cell types, including stem cells (embryonic and adult) (5 –10 ), terminally differentiated somatic cells (i.e., neurons) (7 ,11 ,12 ), muscles (7 ,13 ,14 ), skin (15 ), liver (13 ,16 ,17 ), islet (18 ,19 ), lung epithelium (20 ,21 ), retina (22 ,23 ), primary T lymphocytes (24 ,25 ), and fetal tissues (21 ,26 ) (Table 1 ).

Table 1 Relative Efficiencies of Transduction of Various Cell Lines and Primary Cells with VSV-G Pseudotyped Lentiviral Vectors^a

	Cell type^b	Relative efficiency (+ to ++++^c )
Cell lines
(human)
HEK293	embryonic kidney cells	++++
TE671	rhabdomyosarcoma cells	++++
HeLa	cervical adenocarcinoma	++++
HepG2	hepatoma cells	++++
IB3-1	CF^d bronchial epithelial cells	+++
K562	chronic myelogenous leukemia	++++
U937	myeloid leukemia cells	++++
A20	B cell lymphoma	+
EBV blasts	EBV transformed B lymphoma	+++
(mouse)
Renca	renal cell carcinoma	++++
CT26	colon carcinoma	++++
NIH3T3	fibroblasts	+++
BaF3	pre-B lymphoma	+++
TF1	proerythroblastic leukemia cells	++++
DC2.4	immortalized immature DC	++/+++
P19	embryonic carcinoma	+++
Primary cells
(human)
HUVEC	umbilical vein endothelial cells	+++
CD34⁺ cells	hematopoietic stem/progenitor cells	++/+++^e
monocytes	myeloid cells	++
DC	immature dendritic cells	+++
T cells	lymphocytes	+++^f
B cells (other species)	lymphocytes	+
ESC	mouse embryonic stem cells	+++
neuron	rat neuronal cells	+++
liver stem cells	rat oval cells	+++
liver	primary rat hepatocytes	++
RPE	rat retinal pigment epithelial cells	++++
rod	chicken eye rod cells	++
bone marrow	mouse BM mixed population	+++
HSC	mouse hematopoietic stem/progenitor cells	++

^a ^{The results of primary cell transduction are only for reference comparison; they are based on in vivo or ex vivo culture transduced using HIV-1 vectors carrying different reporter genes.} ^{Therefore, the efficiency can vary markedly depending on the type of reporter gene and the infection condition.} ^b ^{If not specifi ed, the cell types are referring to human cells.} ^c ^{Relative efficiency is defi ned as: ++++ = >50% cells transduced at MOI of 1 to 5; +++ = >50% cells transduced at MOI of 5 to 10; ++ = >30% cells transduced at MOI of 10–20; and + = 1–20% cells transduced at MOI>20.} ^d ^{Cystic fibrosis.} ^e ^{Transduction effi ciency of cytokine-stimulated CD34+ stem cells (+++) is higher than that of unstimulated cells (++).} ^f ^{Both naive and activated human T lymphocytes can be transduced relatively easily with lentiviral vectors; the transgene expression level in na�ve T cells, however, is very low and barely above background level.}

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