title: Transplantation of rat embryonic stem cell-derived retinal progenitor cells preserves the retinal structure and function in rat retinal degeneration.
abstract: BACKGROUND
  Degenerative retinal diseases like age-related macular degeneration (AMD) are the leading cause of blindness. Cell transplantation showed promising therapeutic effect for such diseases, and embryonic stem cell (ESC) is one of the sources of such donor cells. Here, we aimed to generate retinal progenitor cells (RPCs) from rat ESCs (rESCs) and to test their therapeutic effects in rat model.
  
  
  METHODS
  The rESCs (DA8-16) were cultured in N2B27 medium with 2i, and differentiated to two types of RPCs following the SFEBq method with modifications. For rESC-RPC1, the cells were switched to adherent culture at D10, while for rESC-RPC2, the suspension culture was maintained to D14. Both RPCs were harvested at D16. Primary RPCs were obtained from P1 SD rats, and some of them were labeled with EGFP by infection with lentivirus. To generate Rax::EGFP knock-in rESC lines, TALENs were engineered to facilitate homologous recombination in rESCs, which were cotransfected with the targeting vector and TALEN vectors. The differentiated cells were analyzed with live image, immunofluorescence staining, flow cytometric analysis, gene expression microarray, etc. RCS rats were used to mimic the degeneration of retina and test the therapeutic effects of subretinally transplanted donor cells. The structure and function of retina were examined.
  
  
  RESULTS
  We established two protocols through which two types of rESC-derived RPCs were obtained and both contained committed retina lineage cells and some neural progenitor cells (NPCs). These rESC-derived RPCs survived in the host retinas of RCS rats and protected the retinal structure and function in early stage following the transplantation. However, the glia enriched rESC-RPC1 obtained through early and longer adherent culture only increased the b-wave amplitude at 4 weeks, while the longer suspension culture gave rise to evidently neuronal differentiation in rESC-RPC2 which significantly improved the visual function of RCS rats.
  
  
  CONCLUSIONS
  We have successfully differentiated rESCs to glia enriched RPCs and retinal neuron enriched RPCs in vitro. The retinal neuron enriched rESC-RPC2 protected the structure and function of retina in rats with genetic retinal degeneration and could be a candidate cell source for treating some degenerative retinal diseases in human trials.
authors.affiliation: Laboratory of Molecular Developmental Biology, Shanghai Jiao Tong University School of Medicine, Room 208, Building 5, 280 South Chongqing Road, Shanghai, 200025, China. zpqu@sibs.ac.cn.
authors.forename: Zepeng
authors.initials: Z
authors.lastname: Qu
authors.affiliation: Laboratory of Molecular Developmental Biology, Shanghai Jiao Tong University School of Medicine, Room 208, Building 5, 280 South Chongqing Road, Shanghai, 200025, China. yguan@sibs.ac.cn.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Guan
authors.affiliation: Laboratory of Molecular Developmental Biology, Shanghai Jiao Tong University School of Medicine, Room 208, Building 5, 280 South Chongqing Road, Shanghai, 200025, China. lcui@sibs.ac.cn.
authors.forename: Lu
authors.initials: L
authors.lastname: Cui
authors.affiliation: ShanghaiTech University School of Life Science and Technology, Shanghai, 201210, China. songjian@shanghaitech.edu.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Song
authors.affiliation: Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, Shanghai, 200031, China. cauli1984@163.com.
authors.forename: Junjie
authors.initials: J
authors.lastname: Gu
authors.affiliation: Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, Shanghai, 200031, China. hzzhao@sibs.ac.cn.
authors.forename: Hanzhi
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Regenerative Medicine, Stem Cell Research Center, and Institute for Nutritional Sciences, Tongji University School of Medicine, Shanghai, 200092, China. xulei@tongji.edu.cn.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Laboratory of Clinical Visual Science of Tongji Eye Institute, Tongji University School of Medicine, 1239 Siping Road, Medical Building, Room 521, Shanghai, 200092, China. lulixia@tongji.edu.cn.|Department of Regenerative Medicine, Stem Cell Research Center, and Institute for Nutritional Sciences, Tongji University School of Medicine, Shanghai, 200092, China. lulixia@tongji.edu.cn.
authors.forename: Lixia
authors.initials: L
authors.lastname: Lu
authors.affiliation: Laboratory of Molecular Developmental Biology, Shanghai Jiao Tong University School of Medicine, Room 208, Building 5, 280 South Chongqing Road, Shanghai, 200025, China. yjin@sibs.ac.cn.|Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, Shanghai, 200031, China. yjin@sibs.ac.cn.|ShanghaiTech University School of Life Science and Technology, Shanghai, 201210, China. yjin@sibs.ac.cn.
authors.forename: Ying
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Department of Ophthalmology of Shanghai Tenth People's Hospital, and Laboratory of Clinical Visual Science of Tongji Eye Institute, Tongji University School of Medicine, 1239 Siping Road, Medical Building, Room 521, Shanghai, 200092, China. gtxu@tongji.edu.cn.|Department of Regenerative Medicine, Stem Cell Research Center, and Institute for Nutritional Sciences, Tongji University School of Medicine, Shanghai, 200092, China. gtxu@tongji.edu.cn.
authors.forename: Guo-Tong
authors.initials: GT
authors.lastname: Xu
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002465:Cell Movement; D002470:Cell Survival; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000637:transplantation; D020869:Gene Expression Profiling; D018390:Gene Targeting; D066298:In Vitro Techniques; D009474:Neurons / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000166:cytology* / Q000601:surgery; D012162:Retinal Degeneration / Q000601:surgery*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D014785:Vision, Ocular
chemical_list: D000943:Antigens, Differentiation