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Zhenwei Qin, Qiuli FU, Lifang Zhang, Ke Yao; Differentiation of pluripotent stem cells to lentoid bodies to model Congenital cataracts. Invest. Ophthalmol. Vis. Sci. 201657(12):.
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© ARVO (1962-2015); The Authors (2016-present)
The pathological mechanisms of congenital cataracts (CCs) with known mutations were well studied using cell and animal models. However, studies on the sporadic cases which are most of CCs were limited by the lack of appropriate models. Our group modified the induction method of lentoid body (LB) from human urine derived induced pluripotent stem cells(iPSCs) and generated function LBs from iPSCs (unpublished data and presented in ARVO 2015), this study is aim to generate patient specific LBs and explore the possibility of the LBs in modeling the cataract process.
Human urinary cells were collected from the patient with sporadic cataract and prepared for iPSCs induction. Then, the isolated urinary cells were infected with four Yamanaka factors to generate iPSCs. LBs derived from patient specific iPSCs, embryonic stem cells (ESCs) and iPSCs from healthy donor were generated using the established “fried egg” method. Comparison of the inducting process of LBs derived from the three types of cells described above were then investigated through microscopy examination. The transparent degree of three types of LBs were evaluated through MATLAB software using which pixel gray-values of each LBs were calculated.
The patient specific iPSCs were observed to be positive for stem cells markers including Oct4, Sox2, Nanog and Trai1-81. Furthermore, the teratoma assay confirmed the differentiation ability of the iPSC cells. Compared with the LBs derived from ESCs and iPSCs from heathy donor showing transparency morphologies, those derived from the patient-specific iPSCs were blurry. Interestingly, the patient-specific LB exhibited similar morphologies to the LBs from ESCs and healthy iPSCs until D14 when their boundary became blurred. Finally, the LBs with cord-like structure and non-transparency outfit were observed at D25 when the LBs derived from the other cells showed lens-like characteristics. A significant decrease of pixel gray-value was observed in patient-specific LBs (79.1±5.9) compared to normal LBs (135.3±9.2, p=0.001) and ESC LBs (151.9±7.7, p=0.001).
Our results provide evidence that patient-specific congenital cataract disease model could be generated in vitro, thereby suggested a new system to study all kinds of cataract. Pixel gray-value calculation of LB by MATLAB is a novel and feasible quantitative evaluation for LB.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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