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Generation of induced pluripotent stem cells from human Tenon's capsule fibroblasts.
- Source :
-
Molecular vision [Mol Vis] 2012; Vol. 18, pp. 2871-81. Date of Electronic Publication: 2012 Nov 30. - Publication Year :
- 2012
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Abstract
- Purpose: 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.<br />Methods: 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.<br />Results: 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.<br />Conclusions: 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.
- Subjects :
- Adolescent
Adult
Alkaline Phosphatase metabolism
Biomarkers metabolism
Cell Proliferation drug effects
Fibroblasts drug effects
Fibroblasts metabolism
Flow Cytometry
Germ Layers cytology
Humans
Induced Pluripotent Stem Cells drug effects
Induced Pluripotent Stem Cells metabolism
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors pharmacology
Microscopy, Phase-Contrast
Middle Aged
Octamer Transcription Factor-3 pharmacology
Primary Cell Culture
Proto-Oncogene Proteins c-myc pharmacology
Real-Time Polymerase Chain Reaction
SOXB1 Transcription Factors pharmacology
Tenon Capsule drug effects
Tenon Capsule metabolism
Cell Differentiation drug effects
Fibroblasts cytology
Induced Pluripotent Stem Cells cytology
Tenon Capsule cytology
Subjects
Details
- Language :
- English
- ISSN :
- 1090-0535
- Volume :
- 18
- Database :
- MEDLINE
- Journal :
- Molecular vision
- Publication Type :
- Academic Journal
- Accession number :
- 23233789