Photoreceptor Protection by Iris Pigment Epithelial Transplantation Transduced with AAV-Mediated Brain-Derived Neurotrophic Factor Gene

Hojo, M.; Abe, Toshiaki; Sugano, Eriko; Yoshioka, Yuki; Saigo, Yoko; Tomita, Hiroshi; Wakusawa, Ryosuke; Tamai, Makoto

doi:10.1167/iovs.04-0059

Cited by 26 publications

(10 citation statements)

References 40 publications

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“…Our long-term goal is to develop a treatment for patients with AMD or RP. Together with the previous reports that BDNF-expressed IPE cells support the survival of the retinal cells, 15,58 our report shows that this technique of transplanting autologous IPE cells transfected with an appropriate growth factor gene via rAAV2 is a useful method of treating retinal degenerative diseases.…”

Section: Figuresupporting

confidence: 84%

Establishment of Effective Methods for Transducing Genes into Iris Pigment Epithelial Cells by Using Adeno-associated Virus Type 2

Sugano

Tomita

Ishiguro

et al. 2005

Invest. Ophthalmol. Vis. Sci.

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Section: Figuresupporting

confidence: 84%

Establishment of Effective Methods for Transducing Genes into Iris Pigment Epithelial Cells by Using Adeno-associated Virus Type 2

Sugano

Tomita

Ishiguro

et al. 2005

Invest. Ophthalmol. Vis. Sci.

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“…Further studies are needed to confirm effects of phagocytic activities in vivo. Our final goal is to protect photoreceptor cells by the subretinal transplantation of iris pigment epithelial (IPE) cells transduced by using a BDNF gene (Hojo et al, 2004). Changes in phagocytic activities reported here might provide keys to our final goal.…”

Section: Discussionmentioning

confidence: 98%

BDNF Increases the Phagocytic Activity in Cultured Iris Pigment Epithelial Cells

Yoshida

Tomita

Sugano

et al. 2008

Cell Struct. Funct.

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ABSTRACT. To investigate the effect of brain derived neurotrophic factor (BDNF) on the phagocytic activity in iris pigment epithelial (IPE) cells, purified porcine photoreceptor outer segments (POS) were applied to cultured IPE cells for three hours. To measure phagocytic activities, bound and total POS were differentially stained using a double immunofluorescence staining method. BDNF increased the binding of POS in IPE cells in a dose-dependent manner. Ingestion of POS, however, was not affected throughout the concentrations used in this study. To investigate the signal transduction pathways of BDNF, a phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002, and MAPK/ERK kinase (MEK) inhibitor, PD98059, were used for this study. LY294002 had no effect on the binding and ingestion of POS in BDNF-treated IPE cells. On the other hand, PD98059 completely inhibited the increase of POS binding in BDNF-treated cells and also decreased the ingestion of POS. These results indicate that increased POS binding activity by BDNF and the decreased ingestion of POS were mediated through the MAPK pathway.

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“…Cultured cells were incubated with 20% goat serum in PBS and then incubated with the different primary antibodies (Table 1) overnight at 4°C followed by a PBS wash. The cells were then incubated for 60 min with a fluorescence-conjugated secondary antibody followed by a PBS wash, and nuclei were then counterstained with Hoechst 33342 (SigmaAldrich) or propidium iodide (PI; Sigma-Aldrich) according to previous protocols (Hojo et al 2004). BrdU labeling was performed according to protocols used by Hojo et al (2004).…”

Section: Immunocytochemistrymentioning

confidence: 99%

“…The cells were then incubated for 60 min with a fluorescence-conjugated secondary antibody followed by a PBS wash, and nuclei were then counterstained with Hoechst 33342 (SigmaAldrich) or propidium iodide (PI; Sigma-Aldrich) according to previous protocols (Hojo et al 2004). BrdU labeling was performed according to protocols used by Hojo et al (2004). Fluorescence was visualized with either a Leica microscope (DMIRE2, Leica Microsystems) or a confocal microscope (Leica TCS NT; Leica Microsystems).…”

Section: Immunocytochemistrymentioning

confidence: 99%

Characteristics of retinal stem cells from rat optic cup at embryonic day 12.5 (tailbud stage)

2008

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Photoreceptor loss causes irreversible blindness in many retinal diseases. Repair of such damage by cell transplantation is one of the most feasible types of central nervous system treatment. Retinal stem cells (RSC) are a substrate for cell-replacement therapy, and previous studies have shown that RSCs from different developmental stages have distinct properties in proliferative capacity and differentiation potential. The tailbud stage is of special interest in retinogenesis, because RSCs commence differentiation after this period. However, no information about the characteristics of RSCs from the tailbud stage is available. In this study, the characteristics of cell cultures from the rat optic cup (referred to as optic-cup-derived RSCs; OC-RSCs) at embryonic day 12.5 (tailbud stage) were analyzed. OC-RSCs grew either as monolayers or as neurospheres in the presence of basic fibroblast growth factor and could be dissociated into a single cell suspension. Using the MTT assay, immunochemistry, cytogenetic analysis, and flow cytometry, we found that OC-RSCs were easily enriched to 92% by three passages, had a normal diploid karyotype, and exhibited no obvious differences in proliferative rate during eight passages (doubling time: 36 h). OC-RSCs produced retinal specific cells after the addition of serum to the medium, but the differentiation potential was affected by serum concentration. Preliminary results showed that transplanted OC-RSCs were incorporated into the degenerated retina of RCS rats and differentiated into rhodopsin-positive cells. Thus, OC-RSCs, after suitable enrichment, provide a population of stem cells with distinct growth and differentiation properties that make them suitable for research into RSC differentiation and transplantation.

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Photoreceptor Protection by Iris Pigment Epithelial Transplantation Transduced with AAV-Mediated Brain-Derived Neurotrophic Factor Gene

Abstract: Transplantation of AAV2-BDNF-IPE cells may be an alternative method of delivering neurotrophic factors to the lesion.

Cited by 26 publications

References 40 publications

Establishment of Effective Methods for Transducing Genes into Iris Pigment Epithelial Cells by Using Adeno-associated Virus Type 2

Establishment of Effective Methods for Transducing Genes into Iris Pigment Epithelial Cells by Using Adeno-associated Virus Type 2

BDNF Increases the Phagocytic Activity in Cultured Iris Pigment Epithelial Cells

Characteristics of retinal stem cells from rat optic cup at embryonic day 12.5 (tailbud stage)

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