title: Generation of Functional Lentoid Bodies From Human Induced Pluripotent Stem Cells Derived From Urinary Cells.
abstract: The pathological mechanisms underlying cataract formation remain largely unknown on account of the lack of appropriate in vitro cellular models. The aim of this study is to develop a stable in vitro system for human lens regeneration using pluripotent stem cells.
  
  
  
  Isolated human urinary cells were infected with four Yamanaka factors to generate urinary human induced pluripotent stem cells (UiPSCs), which were induced to differentiate into lens progenitor cells and lentoid bodies (LBs). The expression of lens-specific markers was examined by real-time PCR, immunostaining, and Western blotting. The structure and magnifying ability of LBs were investigated using transmission electron microscopy and observing the magnification of the letter "X," respectively.
  
  
  
  We developed a "fried egg" differentiation method to generate functional LBs from UiPSCs. The UiPSC-derived LBs exhibited crystalline lens-like morphology and a transparent structure and expressed lens-specific markers αA-, αB-, β-, and γ-crystallin and MIP. During LB differentiation, the placodal markers SIX1, EYA1, DLX3, PAX6, and the specific early lens markers SOX1, PROX1, FOXE3, αA-, and αB-crystallin were observed at certain time points. Microscopic examination revealed the presence of lens epithelial cells adjacent to the lens capsule as well as both immature and mature fiber-like cells. Optical analysis further demonstrated the magnifying ability (1.7×) of the LBs generated from UiPSCs.
  
  
  
  Our study provides the first evidence toward generating functional LBs from UiPSCs, thereby establishing an in vitro system that can be used to study human lens development and cataractogenesis and perhaps even be useful for drug screening.
authors.affiliation: Eye Center of the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China 2Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.
authors.forename: Qiuli
authors.initials: Q
authors.lastname: Fu
authors.affiliation: Eye Center of the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China 2Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.
authors.forename: Zhenwei
authors.initials: Z
authors.lastname: Qin
authors.affiliation: Eye Center of the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China 2Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.
authors.forename: Xiuming
authors.initials: X
authors.lastname: Jin
authors.affiliation: Eye Center of the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China 2Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.
authors.forename: Lifang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Environmental and Occupational Health, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, Zhejiang Province, China.
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Institute of Environmental Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Jiliang
authors.initials: J
authors.lastname: He
authors.affiliation: Center of Stem Cell and Regenerative Medicine, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China 6Zhejiang Provincial Key Laboratory of Tissue Engineering and Regenerative Medicine, Hangzhou, China.
authors.forename: Junfeng
authors.initials: J
authors.lastname: Ji
authors.affiliation: Eye Center of the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China 2Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Yao
mesh_terms: D015153:Blotting, Western; D002386:Cataract / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002454:Cell Differentiation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000473:pathology; D007908:Lens, Crystalline / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics*
chemical_list: D005136:Eye Proteins; D012333:RNA, Messenger