N.EPSILON.-(Carboxymethyl)lysine Proliferated CD34+ Cells from Rat Choroidal Explant in Culture

Kobayashi, Shogo; Shinohara, Harumichi; Tsuneki, Hiroshi; Nagai, Ryoji; Horiuchi, Seikoh

doi:10.1248/bpb.27.1382

Cited by 9 publications

(31 citation statements)

References 40 publications

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“…22) CML adduct increases the number of CD34 ϩ cells to 67% of total cells. 16) Electron microscope sections of microvessellike structures show additional evidences that narrow lumina were occasionally surrounded by attenuated cellular extensions. The attenuated cell has numerous membrane-bounded polymorphic vesicles.…”

Section: Culture Of Choroidal Explantsmentioning

confidence: 98%

“…[15][16][17] Blood vessels, connective and fatty tissues in the outside of sclera in eye balls of these rats were removed in Dulbecco's modified Eagle's medium (DMEM, Nissui, Tokyo) containing 10% heat-inactive fetal bovine serum (FBS, JRH Bioscience, Lenexa, KS, U.S.A.), 160 U/ml benzyl penicillin potassium (Banyu Seiyaku, Tokyo) and 0.1 mg/ml streptomycin sulfate (Meiji Seika, Tokyo). After removal of the cornea, lens, corpus vitreum and retina from the eye balls, similar size of explants of choroidal tissues were carefully isolated with forceps from the interior aspect of the sclera in 10% FBS-DMEM in the presence of antibiotics, using a surgical microscope.…”

Section: Overproduction Of N E E -(Carboxymethyl)lysine-induced Neovamentioning

confidence: 99%

“…[15][16][17] The similar size of explants were plated on fibrin gels prepared by mixing 3 mg fibrinogen (0.3 ml, Sigma), and 1 U thrombin (Sigma) per ml DMEM containing antibiotics in 16-mm dish (Corning, Corning, NY, U.S.A.). The same volumes of a mixture of the above concentrations of fibrinogen and thrombin solutions were overlaid and solidified.…”

Section: Culture Of Choroidal Explantsmentioning

confidence: 99%

“…Typical photographs of the microvessel-like structures have been published previously. [15][16]21) Thirtynine % of total cells composed in the microvessel-like structures were positive to the antibody against CD34, a marker of vascular endothelial cell and endothelial progenitor cell. 22) CML adduct increases the number of CD34 ϩ cells to 67% of total cells.…”

Section: Culture Of Choroidal Explantsmentioning

confidence: 99%

“…These observations indicated that the microvessel-like structures have a feature of immature microvessel consisted of vascular endothelial progenitor cells such as angioblast. 16) Measurement of VEGF, TNF a a and PDGF-B Contents in Media Cultured with Choroidal Explants by Enzymelinked Immunosorbent Assay Choroidal explants in aged and young rats were cultured with 1% FBS-or 5% FBS-DMEM in the presence or absence of CML-HSA (1 mg/ml) for 6 d. Their media during 0-2, 2-4 or 4-6 d in culture were harvested and stored at Ϫ20°C. The concentrations of immunoreactive VEGF, TNF a and PDGF-B in the culture media were determined by colorimetric competitive enzymelinked immunosorbent assays (ELISA) for mouse VEGF, rat TNF a and human PDGF-B (R&D system), respectively.…”

Section: Culture Of Choroidal Explantsmentioning

confidence: 99%

See 4 more Smart Citations

Overproduction of N.EPSILON.-(Carboxymethyl)lysine-Induced Neovascularization in Cultured Choroidal Explant of Aged Rat

Kobayashi

Nomura

Nishioka

et al. 2007

Biological & Pharmaceutical Bulletin

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Section: Culture Of Choroidal Explantsmentioning

confidence: 98%

Section: Overproduction Of N E E -(Carboxymethyl)lysine-induced Neovamentioning

confidence: 99%

Section: Culture Of Choroidal Explantsmentioning

confidence: 99%

Section: Culture Of Choroidal Explantsmentioning

confidence: 99%

Section: Culture Of Choroidal Explantsmentioning

confidence: 99%

See 3 more Smart Citations

Overproduction of N.EPSILON.-(Carboxymethyl)lysine-Induced Neovascularization in Cultured Choroidal Explant of Aged Rat

Kobayashi

Nomura

Nishioka

et al. 2007

Biological & Pharmaceutical Bulletin

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Injury-Mediated Vascular Regeneration Requires Endothelial ER71/ETV2

Park¹,

Lee²,

Bhang³

et al. 2016

ATVB

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Objective Comprehensive understanding of the mechanisms regulating angiogenesis might provide new strategies for angiogenic therapies for treating diverse physiologic and pathologic ischemic conditions. The ETS factor ETV2 (aka ER71) is essential for the formation of hematopoietic and vascular systems. Despite its indispensable function in vessel development, ETV2 role in adult angiogenesis has not yet been addressed. We have therefore investigated the role of ETV2 in vascular regeneration. Approach and Results We utilized endothelial Etv2 conditional knockout (CKO) mice and ischemic injury models to assess the role of ETV2 in vascular regeneration. While Etv2 expression was not detectable under steady state conditions, its expression was readily observed in endothelial cells following injury. Mice lacking endothelial Etv2 displayed impaired neovascularization in response to eye injury, wounding, or hindlimb ischemic injury. Lentiviral Etv2 expression in ischemic hindlimbs led to improved recovery of blood flow with enhanced vessel formation. Following injury, Flk1 expression and neovascularization were significantly upregulated by Etv2, while Flk1 expression and VEGF response were significantly blunted in Etv2 deficient endothelial cells. Conversely, enforced Etv2 expression enhanced VEGF mediated endothelial sprouting from embryoid bodies. Lentiviral Flk1 expression rescued angiogenesis defects in endothelial Etv2 CKO mice following hindlimb ischemic injury. Furthermore, Etv2+/−; Flk1+/− double heterozygous mice displayed a more severe hindlimb ischemic injury response compared to Etv2+/− or Flk1+/− heterozygous mice, revealing an epistatic interaction between ETV2 and FLK1 in vascular regeneration. Conclusion Our study demonstrates a novel obligatory role for the ETV2 in postnatal vascular repair and regeneration.

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Overproduction of N.EPSILON.-(Carboxymethyl)lysine-Induced Neovascularization in Cultured Choroidal Explant of Streptozotocin-Diabetic Rat

Kobayashi

Suzuki

Tsuneki

et al. 2004

Biological & Pharmaceutical Bulletin

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Action of Ne e -(carboxymethyl)lysine (CML) adduct, an advanced glycation end product, was investigated on neovascularization of cultured choroidal explants in streptozotocin (STZ)-diabetic rat. The choroidal explants of early (4 weeks after an injection of 60 mg/kg STZ) and advanced (8 months after the STZ injection) diabetic rats, and age-matched normal rats were cultured in fibrin gel with Dulbecco's modified Eagle medium containing fetal bovine serum. The number of budded microvessel-like structures was counted and used as an index of in vitro neovascularization. Choroidal explants in the early diabetic stage released vascular endothelial growth factor (VEGF) and tended to increase tumor necrosis factor (TNF) a a and platelet-derived growth factor (PDGF)-B, and concomitantly facilitated growth of sprout and buds, compared to the normal control. When choroidal explants were stimulated with CML-human serum albumin (HSA), its releasing effect was in the order VEGFϾTNFa aϾPDGF-B. CML-HSA and CML-bovine serum albumin augmented the neovascularization in the cultured diabetic explant and their actions did not virtually differ. A monoclonal anti-CML antibody (6D12) inhibited the neovascularization in the advanced diabetes greater than that in the early diabetes. Inhibitory actions of anti-VEGF and anti-TNFa a antibodies on the neovascularization were similar to that of the anti-CML antibody in the diabetes. In conclusion, CML adducts were accumulated and over-produced the actions of VEGF, TNFa a and PDGF-B in the choroidal explant during diabetes in an age-dependent manner. TNFa a and VEGF are likely to play a predominant role for the CML-induced choroidal neovascularization.

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N.EPSILON.-(Carboxymethyl)lysine Proliferated CD34+ Cells from Rat Choroidal Explant in Culture

Cited by 9 publications

References 40 publications

Overproduction of N.EPSILON.-(Carboxymethyl)lysine-Induced Neovascularization in Cultured Choroidal Explant of Aged Rat

Overproduction of N.EPSILON.-(Carboxymethyl)lysine-Induced Neovascularization in Cultured Choroidal Explant of Aged Rat

Injury-Mediated Vascular Regeneration Requires Endothelial ER71/ETV2

Overproduction of N.EPSILON.-(Carboxymethyl)lysine-Induced Neovascularization in Cultured Choroidal Explant of Streptozotocin-Diabetic Rat

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