Abstract
The ocular lens serves as an excellent system to investigate the intricate details of development and differentiation. Generation of lentoid bodies or lens-like structures using pluripotent stem cells is important for understanding the processes critical for lens morphogenesis and the mechanism of cataractogenesis. We previously reported the generation of peripheral blood mononuclear cell (PBMC)-originated, induced pluripotent stem cells (iPSCs). Here, we report generation of lentoid bodies from human embryonic stem cells (hESCs) and (PBMC)-originated, iPSCs employing the “fried egg” method with brief modifications. The ultrastructure analysis of hESC- and iPSC-derived lentoid bodies identified closely packed lens epithelial- and differentiating fiber-like cells. In addition, we performed RNA sequencing (RNA-Seq) based transcriptome profiling of hESC- and iPSC-derived lentoid bodies at differentiation day 25. Next-generation RNA sequencing (RNA-Seq) of hESC- and iPSC-derived lentoid bodies detected expression (≥0.659 RPKM) of 13,975 and 14,003 genes, respectively. Comparative transcriptome analysis of hESC- and iPSC-derived lentoid bodies revealed 13,563 (>96%) genes common in both datasets. Among the genes common in both transcriptome datasets, 12,856 (~95%) exhibited a quantitatively similar expression profile. Next, we compared the mouse lens epithelial and fiber cell transcriptomes with hESC- and iPSC-derived lentoid bodies transcriptomes and identified > 96% overlap with lentoid body transcriptomes. In conclusion, we report first-time comparative transcriptome analysis of hESC- and iPSC-derived lentoid bodies at differentiation day 25.
Highlights
The ocular lens serves as an excellent system to investigate the intricate details of development and differentiation
The flow cytometry analysis confirmed that PBMCoriginated, induced pluripotent stem cells (iPSCs) were positive for SSEA4 (94.56 ± 2.43%) and TRA-1-60 (89.36 ± 1.57%) (Supplementary Fig. 1c,d)
The use of patient-specific iPSC-derived lentoid bodies may provide a handful tool for understanding the process of cataractogenesis and possible therapeutic intervention
Summary
The ocular lens serves as an excellent system to investigate the intricate details of development and differentiation. We report generation of lentoid bodies from human embryonic stem cells (hESCs) and (PBMC)-originated, iPSCs employing the “fried egg” method with brief modifications. Differentiation of pluripotent stem cells to lentoid bodies or lens-like structures is important for understanding lens development and investigating the processes critical for lens morphogenesis. Fu and colleagues developed lentoid bodies from human urinary epithelial cell-originated, iPSCs employing the “fried egg” method of differentiation[4]. These lentoid bodies expressed lens-associated markers and displayed transparent structure like a human lens[4]. We report RNA sequencing (RNA-Seq) based transcriptome profiling of lentoid bodies derived from hESCs and peripheral blood mononuclear cell (PBMC)-originated, iPSCs
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