PURPOSE: Human embryonic stem cell-derived RPE (hES-RPE) transplantation is a promising therapy for atrophic age-related macular degeneration (AMD); however, future therapeutic approaches may consider co-transplantation of hES-RPE with retinal progenitor cells (RPCs) as a replacement source for lost photoreceptors. The purpose of this study was to determine the effect of polarization of hES-RPE monolayers on their ability to promote survival of RPCs. METHODS: The hES-3 cell line was used for derivation of RPE. Polarization of hES-RPE was achieved by prolonged growth on permeable inserts. RPCs were isolated from 16- to 18-week-gestation human fetal eyes. ELISA was performed to measure pigment epithelium-derived factor (PEDF) levels from conditioned media. RESULTS: Pigmented RPE-like cells appeared as early as 4 weeks in culture and were subcultured at 8 weeks. Differentiated hES-RPE had a normal chromosomal karyotype. Phenotypically polarized hES-RPE cells showed expression of RPE-specific genes. Polarized hES-RPE showed prominent expression of PEDF in apical cytoplasm and a marked increase in secretion of PEDF into the medium compared with nonpolarized culture. RPCs grown in the presence of supernatants from polarized hES-RPE showed enhanced survival, which was ablated by the presence of anti-PEDF antibody. CONCLUSIONS: hES-3 cells can be differentiated into functionally polarized hES-RPE cells that exhibit characteristics similar to those of native RPE. On polarization, hES-RPE cells secrete high levels of PEDF that can support RPC survival. These experiments suggest that polarization of hES-RPE would be an important feature for promotion of RPC survival in future cell therapy for atrophic AMD.
PURPOSE:Human embryonic stem cell-derived RPE (hES-RPE) transplantation is a promising therapy for atrophic age-related macular degeneration (AMD); however, future therapeutic approaches may consider co-transplantation of hES-RPE with retinal progenitor cells (RPCs) as a replacement source for lost photoreceptors. The purpose of this study was to determine the effect of polarization of hES-RPE monolayers on their ability to promote survival of RPCs. METHODS: The hES-3 cell line was used for derivation of RPE. Polarization of hES-RPE was achieved by prolonged growth on permeable inserts. RPCs were isolated from 16- to 18-week-gestation human fetal eyes. ELISA was performed to measure pigment epithelium-derived factor (PEDF) levels from conditioned media. RESULTS: Pigmented RPE-like cells appeared as early as 4 weeks in culture and were subcultured at 8 weeks. Differentiated hES-RPE had a normal chromosomal karyotype. Phenotypically polarized hES-RPE cells showed expression of RPE-specific genes. Polarized hES-RPE showed prominent expression of PEDF in apical cytoplasm and a marked increase in secretion of PEDF into the medium compared with nonpolarized culture. RPCs grown in the presence of supernatants from polarized hES-RPE showed enhanced survival, which was ablated by the presence of anti-PEDF antibody. CONCLUSIONS:hES-3 cells can be differentiated into functionally polarized hES-RPE cells that exhibit characteristics similar to those of native RPE. On polarization, hES-RPE cells secrete high levels of PEDF that can support RPC survival. These experiments suggest that polarization of hES-RPE would be an important feature for promotion of RPC survival in future cell therapy for atrophic AMD.
Authors: Jayakrishna Ambati; Balamurali K Ambati; Sonia H Yoo; Sean Ianchulev; Anthony P Adamis Journal: Surv Ophthalmol Date: 2003 May-Jun Impact factor: 6.048
Authors: Brian J Raisler; Kenneth I Berns; Maria B Grant; Denis Beliaev; William W Hauswirth Journal: Proc Natl Acad Sci U S A Date: 2002-06-18 Impact factor: 11.205
Authors: M Miyazaki; Y Ikeda; Y Yonemitsu; Y Goto; T Sakamoto; T Tabata; Y Ueda; M Hasegawa; S Tobimatsu; T Ishibashi; K Sueishi Journal: Gene Ther Date: 2003-08 Impact factor: 5.250
Authors: Niina Onnela; Virpi Savolainen; Kati Juuti-Uusitalo; Hanna Vaajasaari; Heli Skottman; Jari Hyttinen Journal: Med Biol Eng Comput Date: 2011-12-22 Impact factor: 2.602
Authors: Tim U Krohne; Peter D Westenskow; Toshihide Kurihara; David F Friedlander; Mandy Lehmann; Alison L Dorsey; Wenlin Li; Saiyong Zhu; Andrew Schultz; Junhua Wang; Gary Siuzdak; Sheng Ding; Martin Friedlander Journal: Stem Cells Transl Med Date: 2012-02 Impact factor: 6.940
Authors: Anni Sorkio; Heidi Hongisto; Kai Kaarniranta; Hannu Uusitalo; Kati Juuti-Uusitalo; Heli Skottman Journal: Tissue Eng Part A Date: 2014-01-20 Impact factor: 3.845
Authors: Zala Lužnik; Mohit Parekh; Marina Bertolin; Carlo Griffoni; Diego Ponzin; Stefano Ferrari Journal: Stem Cells Transl Med Date: 2015-06-01 Impact factor: 6.940
Authors: Ratnesh K Singh; Ramya K Mallela; Pamela K Cornuet; Aaron N Reifler; Andrew P Chervenak; Michael D West; Kwoon Y Wong; Igor O Nasonkin Journal: Stem Cells Dev Date: 2015-09-10 Impact factor: 3.272