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

Mol Vis. 2012:18:2871-81. Epub 2012 Nov 30.

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.

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.

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.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Alkaline Phosphatase / metabolism
  • Biomarkers / metabolism
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects
  • Fibroblasts / cytology*
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Flow Cytometry
  • Germ Layers / cytology
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • 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 / cytology*
  • Tenon Capsule / drug effects
  • Tenon Capsule / metabolism

Substances

  • Biomarkers
  • KLF4 protein, human
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors
  • MYC protein, human
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Proto-Oncogene Proteins c-myc
  • SOXB1 Transcription Factors
  • Alkaline Phosphatase