Efficient Generation of Lens Progenitor Cells from Cataract Patient–Specific Induced Pluripotent Stem Cells

Qiu, Xiaodi; Yang, Jin; Liu, Tianjin; Jiang, Yongxiang; Le, Qihua; Lü, Yi

doi:10.1371/journal.pone.0032612

Cited by 50 publications

(43 citation statements)

References 40 publications

(50 reference statements)

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“…Although iPSCs have been previously generated from the lens epithelial cells of cataract patients, no proper cataract model has been established. 25 Our study opens up the possibility of generating patient-specific cataract models for the evaluation of the efficacy of these compounds.…”

Section: Discussionmentioning

confidence: 93%

“…24 Human ESC-/iPSC-derived LBs have also been reported by several groups. 22,25,26 However, there are certain limitations with regard to the size and morphology of the LBs generated with the methods reported so far. The maximum diameter of the LBs induced in our study reached approximately 3 mm, which is, to our knowledge, the largest LBs generated in vitro.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Generation of Functional Lentoid Bodies From Human Induced Pluripotent Stem Cells Derived From Urinary Cells

Qin

Jin

et al. 2017

Invest. Ophthalmol. Vis. Sci.

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Citation: Fu Q, Qin Z, Jin X, et al. Generation of functional lentoid bodies from human induced pluripotent stem cells derived from urinary cells. Invest Ophthalmol Vis Sci. 2017;58:517-527. DOI:10.1167/ iovs.16-20504 PURPOSE. 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. METHODS.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. RESULTS.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 aA-, aB-, b-, and c-crystallin and MIP. During LB differentiation, the placodal markers SIX1, EYA1, DLX3, PAX6, and the specific early lens markers SOX1, PROX1, FOXE3, aA-, and aB-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 (1.73) of the LBs generated from UiPSCs. CONCLUSIONS.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.

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Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Generation of Functional Lentoid Bodies From Human Induced Pluripotent Stem Cells Derived From Urinary Cells

Qin

Jin

et al. 2017

Invest. Ophthalmol. Vis. Sci.

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“…Although presently there is no evidence of differentiation of these cells into a fiber cell fate, it will be interesting to investigate the effect of conditions used in studies that describe directed differentiation of embryonic stem cell and induced pluripotent stem cells into lens fate (Qiu et al, 2012; Yang et al, 2010). Nevertheless, these LECs offer advantages over primary cell culture that pose challenges such as potential variability between samples or limited lifespan, and in past have been useful in identifying targets that were later validated in the lens tissue (Lachke et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of mouse lens epithelial cell lines and their suitability to study RNA granules and cataract associated genes

Terrell

Anand

Smith

et al. 2015

Experimental Eye Research

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The discovery of cytosolic RNA granule (RG) component proteins associated with human cataract has initiated investigations on post-transcriptional mechanisms of gene expression control in the lens. Application of established mouse lens epithelial cell lines (LECs) can provide rapid insights on RG function in lens cells, especially because mouse mutants in several RG components are not available. However, although these LECs represent potential reagents for such analyses, they are uncharacterized for lens gene expression or RG formation. Therefore, a detailed molecular and cellular characterization of three permanent mouse LECs 17EM15, 21EM15 and αTN4 is performed in this study. Comparative analysis between microarray gene expression datasets on LEC 21EM15 and iSyTE lens tissue demonstrates that 30% of top 200 iSyTE identified lens-enriched genes are expressed in these cells. Majority of these candidates are independently validated to either have lens expression, function or linkage to cataract. Moreover, analysis of microarray data with genes described in Cat-Map, an online database of cataract associated genes and loci, demonstrates that 131 genes linked to cataract loci are expressed in 21EM15 cells. Furthermore, gene expression in LECs is compared to isolated lens epithelium or fiber cells by qRT-PCR and by comparative analyses with publically available epithelium or fiber-specific microarray and RNA-seq (sequencing) datasets. Expression of select candidate genes was validated by regular and real-time quantitative RT-PCR. Expression of lens epithelium-enriched genes Foxe3, Pax6, Anxa4 and Mcm4 is up-regulated in LEC lines, compared to isolated lens fiber cells. Moreover, similar to isolated lens epithelium, all three LECs exhibit down-regulation of fiber cell-expressed genes Crybb1, Mip and Prox1 when compared to fiber cells. These data indicate that the LEC lines exhibit greater similarity to lens epithelium than to fiber cells. Compared to non-lens cell line NIH3T3, LECs exhibit significantly enriched expression of transcription factors with important function in the lens, namely Pax6, Foxe3 and Prox1. In addition to these genes, all three LECs also express key lens- and cataract-associated genes, namely Dkk3, Epha2, Hsf4, Jag1, Mab21l1, Meis1, Pknox1, Pou2f1, Sfrp1, Sparc, Tdrd7 and Trpm3. Additionally, 21EM15 microarrays indicate expression of Chmp4b, Cryab and Tcfap2a among others important genes. Immunostaining with makers for Processing bodies (P-bodies) and Stress granules (SGs) demonstrates that these classes of RGs are robustly expressed in all three LECs. Moreover, under conditions of stress, 17EM15 and αTN4 exhibit significantly higher numbers of P-bodies and SGs compared to NIH3T3 cells. In sum, these data indicate that mouse LECs 21EM15, 17EM15 and αTN4 express key lens or cataract genes, are similar to lens epithelium than fiber cells, and exhibit high levels of P-bodies and SGs, indicating their suitability for investigating gene expression control and RG function in lens-derived cells.

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“…The lens stem cells might thus reside in the lens capsule [47,48]. Lens stem cells have not yet been identified.…”

Section: Lensmentioning

confidence: 99%

Ocular stem cells: a status update!

et al. 2014

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Stem cells are unspecialized cells that have been a major focus of the field of regenerative medicine, opening new frontiers and regarded as the future of medicine. The ophthalmology branch of the medical sciences was the first to directly benefit from stem cells for regenerative treatment. The success stories of regenerative medicine in ophthalmology can be attributed to its accessibility, ease of follow-up and the eye being an immune-privileged organ. Cell-based therapies using stem cells from the ciliary body, iris and sclera are still in animal experimental stages but show potential for replacing degenerated photoreceptors. Limbal, corneal and conjunctival stem cells are still limited for use only for surface reconstruction, although they might have potential beyond this. Iris pigment epithelial, ciliary body epithelial and choroidal epithelial stem cells in laboratory studies have shown some promise for retinal or neural tissue replacement. Trabecular meshwork, orbital and sclera stem cells have properties identical to cells of mesenchymal origin but their potential has yet to be experimentally determined and validated. Retinal and retinal pigment epithelium stem cells remain the most sought out stem cells for curing retinal degenerative disorders, although treatments using them have resulted in variable outcomes. The functional aspects of the therapeutic application of lenticular stem cells are not known and need further attention. Recently, embryonic stem cell-derived retinal pigment epithelium has been used for treating patients with Stargardts disease and age-related macular degeneration. Overall, the different stem cells residing in different components of the eye have shown some success in clinical and animal studies in the field of regenerative medicine.

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Efficient Generation of Lens Progenitor Cells from Cataract Patient–Specific Induced Pluripotent Stem Cells

Cited by 50 publications

References 40 publications

Generation of Functional Lentoid Bodies From Human Induced Pluripotent Stem Cells Derived From Urinary Cells

Generation of Functional Lentoid Bodies From Human Induced Pluripotent Stem Cells Derived From Urinary Cells

Molecular characterization of mouse lens epithelial cell lines and their suitability to study RNA granules and cataract associated genes

Ocular stem cells: a status update!

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