New treatment and functional analysis of age related macular diseases

当从患有年龄相关性黄斑变性（AMD）的患者手术切除脉络膜新生血管（CNV）时，视网膜色素上皮（RPE）也与CNV一起沿着被去除。在这些情况下，RPE细胞移植到视网膜下腔是一种可能的方法来恢复视力。本研究的目的是从AMD患者手术切除的CNV标本中培养RPE细胞，并将RPE细胞片移植到手术切除的视网膜下腔中，2004年，我们进行了几项实验，以解决单层虹膜上皮或RPE细胞片移植到视网膜下腔的技术问题。1)将通过虹膜切除术相对容易获得的兔虹膜上皮或人RPE细胞系（ARPE-19）与添加到培养基中的磁性纳米颗粒一起培养以磁性标记RPE细胞片。贴上磁性标签的纸张可以用铁丝连接到 关于我们 圆柱形钕磁铁。（Tissue Eng，2005）。2)用3 T3-J2细胞作为饲养层培养角膜内皮细胞，以研究更好地保存培养细胞的方法（Cornea，2005）。3)在名古屋大学医学伦理委员会的许可下，用3 T3-J2细胞作为饲养层从手术去除的CNV膜进行RPE细胞培养。对培养的细胞片进行了细胞化学研究，以确认人RPE细胞的选择性生长（Cytotherapy，2005）。在2005年，我们进行了几个实验来研究AMD中新生血管形成的机制，旨在通过将基因转染到培养的RPE细胞片来进行基因治疗。1)CNV的免疫组化研究显示，炎症在AMD的新血管形成中起重要作用（已提交）。2)血清CRP水平与炎性细胞因子基因多态性相关，在AMD和息肉状脉络膜血管病变患者中升高。少

When choroidal neovascularization (CNV) is surgically excised from patients with age related macular degeneration (AMD), the retinal pigment epithelium (RPE) is also removed along with CNV. In those cases, RPE cell transplantation into the subretinal space is a possible approach to restore the vision. The purpose of our study was to cultivate the RPE cells from CNV specimens that are surgically removed from the patients with AMD and transplant the RPE cell sheet into the subretinal space where the RPE is surgically excised.In 2004, we have performed several experiments to solve the technical problems for transplantation of the single layered iris epithelium or RPE cell sheet to the subretinal space. 1)Rabbit iris epithelium, which is relatively easy to obtain by iridectomy, or human RPE cell line (ARPE-19) was cultured with magnetite nanoparticles added to the medium to magnetically label the RPE cell sheet. Sheet labeled magnetically could be handled with an iron wire attached to the … More cylindrical neodymium magnet. (Tissue Eng, 2005). 2)Corneal endothelial cells were cultured with 3T3-J2 cells as a feeder layer to investigate the method to better preserve the cultured cells (Cornea, 2005). 3)With permission by Nagoya University Medical Ethics Committee, RPE cell culture from a surgically removed CNV membrane with 3T3-J2 cells as a feeder layer was performed. Cultured cell sheet were investigated immunohistochemically to confirm selective growth of human RPE cells (Cytotherapy, 2005).In 2005, we have performed several experiments to investigate mechanism of neovascularization in AMD, aiming to perform gene therapy by transfecting gene to cultured RPE cell sheet. 1)Immunohistochemical study of CNV revealed that inflammation plays an important role in neovascularization in AMD (submitted). 2)The serum CRP level has correlation with inflammatory cytokine gene polymorphisms (in preparation), and it is increased in AMD and polypoidal choroidal vasculopathy patients (in preparation). Less