Department of Ophthalmology, Eye and ENT Hospital of Fudan University, Shanghai, People's Republic of China.
PLoS One. 2012;7(3):e32612. doi: 10.1371/journal.pone.0032612. Epub 2012 Mar 5.
The development of a technique to induce the transformation of somatic cells to a pluripotent state via the ectopic expression of defined transcription factors was a transformational event in the field of regenerative medicine. The development of this technique also impacted ophthalmology, as patient-specific induced pluripotent stemcells (iPSCs) may be useful resources for some ophthalmological diseases. The lens is a key refractive element in the eye that focuses images of the visual world onto the retina. To establish a new model for drug screening to treat lens diseases and investigating lens aging and development, we examined whether human lens epithelial cells (HLECs) could be induced into iPSCs and if lens-specific differentiation of these cells could be achieved under defined chemical conditions. We first efficiently reprogrammed HLECs from age-related cataract patients to iPSCs with OCT-4, SOX-2, and KLF-4. The resulting HLEC-derived iPS (HLE-iPS) colonies were indistinguishable from human ES cells with respect to morphology, gene expression, pluripotent marker expression and their ability to generate all embryonic germ-cell layers. Next, we performed a 3-step induction procedure: HLE-iPS cells were differentiated into large numbers of lens progenitor-like cells with defined factors (Noggin, BMP and FGF2), and we determined that these cells expressed lens-specific markers (PAX6, SOX2, SIX3, CRYAB, CRYAA, BFSP1, and MIP). In addition, HLE-iPS-derived lens cells exhibited reduced expression of epithelial mesenchymal transition (EMT) markers compared with human embryonic stem cells (hESCs) and fibroblast-derived iPSCs. Our study describes a highly efficient procedure for generating lens progenitor cells from cataract patient HLEC-derived iPSCs. These patient-derived pluripotent cells provide a valuable model for studying the developmental and molecular biological mechanisms that underlie cell determination in lens development and cataract pathophysiology.
通过异位表达特定转录因子诱导体细胞向多能性状态转化的技术的发展，是再生医学领域的一个变革性事件。这项技术的发展也对眼科学产生了影响，因为患者特异性诱导多能干细胞（iPSC）可能是一些眼科疾病的有用资源。晶状体是眼睛中的关键屈光元件，它将视觉世界的图像聚焦在视网膜上。为了建立一种新的用于治疗晶状体疾病的药物筛选模型，并研究晶状体衰老和发育，我们研究了人晶状体上皮细胞（HLEC）是否可以被诱导成为 iPSC，以及这些细胞在特定的化学条件下是否可以进行晶状体特异性分化。我们首先利用 OCT-4、SOX-2 和 KLF-4 从年龄相关性白内障患者中高效地将 HLEC 重编程为 iPSC。所得的 HLEC 来源的 iPS（HLE-iPS）集落在形态、基因表达、多能性标志物表达及其生成所有胚胎生殖细胞层的能力方面与人类胚胎干细胞（hESC）无法区分。接下来，我们进行了 3 步诱导程序：HLE-iPS 细胞在定义的因子（Noggin、BMP 和 FGF2）下分化为大量晶状体祖细胞样细胞，我们确定这些细胞表达晶状体特异性标志物（PAX6、SOX2、SIX3、CRYAB、CRYAA、BFSP1 和 MIP）。此外，与 hESC 和成纤维细胞来源的 iPSC 相比，HLE-iPSC 衍生的晶状体细胞表现出上皮间质转化（EMT）标志物的表达减少。我们的研究描述了一种从白内障患者 HLEC 来源的 iPSC 中高效生成晶状体祖细胞的方法。这些患者来源的多能细胞为研究晶状体发育和白内障病理生理学中细胞决定的发育和分子生物学机制提供了一个有价值的模型。
