Modeling Keratoconus Using Induced Pluripotent Stem Cells
| Autor: | Roy Joseph, Om P. Srivastava, Roswell R. Pfister |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Adult Keratinocytes Male induced pluripotent stem cells MAP Kinase Signaling System Cellular differentiation keratoconus Oct-4 Biology Stem cell marker Models Biological Cornea 03 medical and health sciences Kruppel-Like Factor 4 Phosphatidylinositol 3-Kinases SOX2 disease modeling Humans Receptor Fibroblast Growth Factor Type 2 Induced pluripotent stem cell Cells Cultured Cell Proliferation Induced stem cells Cell Differentiation Fibroblasts Cell biology 030104 developmental biology MRNA Sequencing FGFR2 Female sense organs Stem cell |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 0146-0404 |
| Popis: | 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 pluripotency markers, SSEA4, Oct4, Sox2, Tra-1-60, and also expressed pax6. Our transcriptome analysis showed 4300 genes, which had 2-fold change and 870 genes with a q-value of |
| Databáze: | OpenAIRE |
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