Differentiation of Induced Pluripotent Stem Cells to Lentoid Bodies Expressing a Lens Cell-Specific Fluorescent Reporter.

Taruna Anand,Wiebke Garrels,Dharmendra Kumar,Rüdiger Behr,Thirumala R Talluri,Ayan Mukherjee,Katharina Debowski,Wilfried A Kues,Joseph Najbauer

doi:10.1371/journal.pone.0157570

Abstract

Curative approaches for eye cataracts and other eye abnormalities, such as myopia and hyperopia currently suffer from a lack of appropriate models. Here, we present a new approach for in vitro growth of lentoid bodies from induced pluripotent stem (iPS) cells as a tool for ophthalmological research. We generated a transgenic mouse line with lens-specific expression of a fluorescent reporter driven by the alphaA crystallin promoter. Fetal fibroblasts were isolated from transgenic fetuses, reprogrammed to iPS cells, and differentiated to lentoid bodies exploiting the specific fluorescence of the lens cell-specific reporter. The employment of cell type-specific reporters for establishing and optimizing differentiation in vitro seems to be an efficient and generally applicable approach for developing differentiation protocols for desired cell populations.

Highlights

Age-related cataracts are one of the most prevalent ocular conditions resulting from the failure of specific cell types and represent the major eye disease in humans [1]
A detailed characterization of the crystallin-promoter driven tdTomato reporter (cryTom) mouse line suggested that the reporter faithfully mirrored the spatial and temporal expression pattern of the Cryaa gene
Induced PS cells could be derived from cryTom fibroblasts; upon exposing the cryTom induced pluripotent stem (iPS) cells to a differentiation protocol, expression of the tdTomato reporter was resumed, allowing simple identification and vital recording of lentoid body growth in vitro

Summary

Introduction

Age-related cataracts are one of the most prevalent ocular conditions resulting from the failure of specific cell types and represent the major eye disease in humans [1]. The lens progenitor cells originate from a vesicle at the lens placode [3,4] and the lens fiber cells terminally differentiate to contributing to the three-dimensional structure of the lens. This includes a massive up-regulation of lens-specific genes, such as alpha- and beta-crystallins [5,6]. Expression of alphaA crystallin (Cryaa) is initiated in the cells of the PLOS ONE | DOI:10.1371/journal.pone.0157570 June 20, 2016

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