|Subretinal Implantation of Retinal Pigment Epithelial Cells Derived From Human Embryonic Stem Cells: Improved Survival When Implanted as a Monolayer|
|Diniz, Bruno1,2; Thomas, Padmaja1; Thomas, Biju1,3; Ribeiro, Ramiro1,2,4; Hu, Yuntao1,5; Brant, Rodrigo1,2; Ahuja, Ashish1; Zhu, Danhong1; Liu, Laura1,6; Koss, Michael1,7; Maia, Mauricio2; Chader, Gerald1; Hinton, David R.1,3,8; Humayun, Mark S.1,3|
|关键词||Age-related Macular Degeneration Retinal Pigment Epithelium Human Embryonic Stem Cells|
|刊名||INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE|
|WOS标题词||Science & Technology|
|关键词[WOS]||HUMAN BRUCHS MEMBRANE ; MACULAR DEGENERATION ; RCS RATS ; IMMUNE-RESPONSES ; VISUAL FUNCTION ; INTACT SHEETS ; ATHYMIC RAT ; NUDE RATS ; TRANSPLANTATION ; MODEL|
PURPOSE. To evaluate cell survival and tumorigenicity of human embryonic stem cell-derived retinal pigment epithelium (hESC-RPE) transplantation in immunocompromised nude rats. Cells were transplanted as a cell suspension (CS) or as a polarized monolayer plated on a parylene membrane (PM).
METHODS. Sixty-nine rats (38 male, 31 female) were surgically implanted with CS (n = 33) or PM (n = 36). Cohort subsets were killed at 1, 6, and 12 months after surgery. Both ocular tissues and systemic organs (brain, liver, kidneys, spleen, heart, and lungs) were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned. Every fifth section was stained with hematoxylin and eosin and analyzed histologically. Adjacent sections were processed for immunohistochemical analysis (as needed) using the following antibodies: anti-RPE65 (RPE-specific marker), anti-TRA-1-85 (human cell marker), anti-Ki67 (proliferation marker), anti-CD68 (macrophage), and anti-cytokeratin (epithelial marker).
RESULTS. The implanted cells were immunopositive for the RPE65 and TRA-1-85. Cell survival (P = 0.006) and the presence of a monolayer (P < 0.001) of hESC-RPE were significantly higher in eyes that received the PM. Gross morphological and histological analysis of the eye and the systemic organs after the surgery revealed no evidence of tumor or ectopic tissue formation in either group.
CONCLUSIONS. hESC-RPE can survive for at least 12 months in an immunocompromised animal model. Polarized monolayers of hESC-RPE show improved survival compared to cell suspensions. The lack of teratoma or any ectopic tissue formation in the implanted rats bodes well for similar results with respect to safety in human subjects.
|项目编号||DR1-01444 ; EY03040 ; BEX 2326-11-6|
|资助机构||California Institute of Regenerative Medicine ; Research to Prevent Blindness ; National Eye Institute ; CAPES Foundation|
|作者单位||1.Doheny Eye Inst, Los Angeles, CA 90033 USA|
2.Hosp Univ Curitiba, Dept Ophthalmol, Curitiba, Parana, Brazil
3.Peking Univ, Hosp 3, Ctr Eye, Beijing 100871, Peoples R China
4.Univ Fed Sao Paulo, Dept Ophthalmol, Sao Paulo, Brazil
5.Univ So Calif, Keck Sch Med, Dept Ophthalmol, Los Angeles, CA 90033 USA
6.Chang Gung Mem Hosp, Dept Ophthalmol, Linkou, Taiwan
7.Goethe Univ Frankfurt, Dept Ophthalmol, D-60054 Frankfurt, Germany
8.Univ So Calif, Keck Sch Med, Dept Ophthalmol, Los Angeles, CA 90033 USA
|Diniz, Bruno,Thomas, Padmaja,Thomas, Biju,et al. Subretinal Implantation of Retinal Pigment Epithelial Cells Derived From Human Embryonic Stem Cells: Improved Survival When Implanted as a Monolayer[J]. INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE,2013,54(7):5087-5096.|
|APA||Diniz, Bruno.,Thomas, Padmaja.,Thomas, Biju.,Ribeiro, Ramiro.,Hu, Yuntao.,...&Humayun, Mark S..(2013).Subretinal Implantation of Retinal Pigment Epithelial Cells Derived From Human Embryonic Stem Cells: Improved Survival When Implanted as a Monolayer.INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE,54(7),5087-5096.|
|MLA||Diniz, Bruno,et al."Subretinal Implantation of Retinal Pigment Epithelial Cells Derived From Human Embryonic Stem Cells: Improved Survival When Implanted as a Monolayer".INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE 54.7(2013):5087-5096.|