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  • Generation of induced pluripotent stem cells from human Tenon's capsule fibroblasts.

Generation of induced pluripotent stem cells from human Tenon's capsule fibroblasts.

Molecular vision (2012-12-13)
Fei Deng, Huiling Hu, Mengfei Chen, Xuerong Sun, Xiaohong Liu, Zhizhang Dong, Ying Liu, Lei Xi, Jing Zhuang, Jian Ge
ABSTRACT

This study aimed to develop a feasible and efficient method for generating embryonic stem cell (ESC)-like induced pluripotent stem (iPS) cells from human Tenon's capsule fibroblasts (HTFs) through the expression of a defined set of transcription factors, which will have significant application value for ophthalmic personalized regenerative medicine. HTFs were harvested from fresh samples, and reprogramming was induced by the exogenous expression of the four classic transcription factors, OCT-3/4, SOX-2, KLF-4, and C-MYC. The HTF-derived iPS (TiPS) cells were analyzed with phase contrast microscopy, real-time PCR, immunofluorescence, FACS analysis, alkaline phosphatase activity analysis, and a teratoma formation assay. Human ESC colonies were used as the positive control. The resulting HTF-derived iPS cell colonies were indistinguishable from human ESC colonies regarding morphology, gene expression levels, pluripotent gene expression, alkaline phosphatase activity, and the ability to generate all three embryonic germ layers. This study presents a simple, efficient, practical procedure for generating patient-tailored iPS cells from HTFs. These cells will serve as a valuable and preferred candidate donor cell population for ophthalmological regenerative medicine.

MATERIALS
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Product Description

Sigma-Aldrich
Quantitative Alkaline Phosphatase ES Characterization Kit, This Quantitative Alkaline Phosphatase Embryonic Stem (ES) Characterization Kit easily quantifies the amount of alkaline phosphatase present within their embryonic stem cell cultures using a convenient 96-well colorimetric enzymatic assay.