Modeling Keratoconus Using Induced Pluripotent Stem Cells.

ABSTRACT:

Purpose

To model keratoconus (KC) using induced pluripotent stem cells (iPSC) generated from fibroblasts of both KC and normal human corneal stroma by a viral method.

Methods

Both normal and KC corneal fibroblasts from four human donors were reprogramed directly by delivering reprogramming factors in a single virus using 2A "self-cleaving" peptides, using a single polycistronic lentiviral vector coexpressing four transcription factors (Oct 4, Sox2, Klf4, and Myc) to yield iPSC. These iPS cells were characterized by immunofluorescence detection using of stem cell markers (SSEA4, Oct4, and Sox2). The mRNA sequencing was performed and the datasets were analyzed using ingenuity pathways analysis (IPA) software.

Results

The generated stem cell-like clones expressed the plu

SUBMITTER: Joseph R 

PROVIDER: S-EPMC5996875 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature