Directing adult human periodontal ligament-derived stem cells to retinal fate

Li Huang, Jiajian Liang, Yiqun Geng, Wai Ming Tsang, Xiaowu Yao, Vishal Jhanji, Mingzhi Zhang, Herman S. Cheung, Chi Pui Pang, Gary Hin Fai Yam

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

PURPOSE. To investigate the retinal fate competence of human postnatal periodontal ligament (PDL)-derived stem cells (PDLSC) through a directed differentiation mimicking mammalian retinogenesis. METHODS. Human teeth were collected from healthy subjects younger than 35 years old. Primary PDLSC were isolated by collagenase digestion and cultivated. PDLSC at passage 3 were cultured in the induction media containing Noggin (antagonist of bone morphogenic protein) and Dkk-1 (antagonist of Wnt/b-catenin signaling). Gene expression of neural crest cells, retinal progenitors, and retinal neurons, including photoreceptors, was revealed by RNA analyses, immunofluorescence, and flow cytometry. The neuronal-like property of differentiated cells in response to excitatory glutamate was examined by fluo-4-acetoxymethyl calcium imaging assay. RESULTS. Primary human PDLSC stably expressed marker genes for neural crest (Notch1, BMP2, Slug, Snail, nestin, and Tuj1), mesenchymal stem cell (CD44, CD90, and vimentin), and embryonic stem cell (c-Myc, Klf4, Nanog, and SSEA4). Under low attachment culture, PDLSC generated neurospheres expressing nestin, p75/NGFR, Pax6, and Tuj1 (markers of neural progenitors). When neurospheres were plated on Matrigel-coated surface, they exhibited rosette-like outgrowth. They expressed eye field transcription factors (Pax6, Rx, Lhx, Otx2). By flow cytometry, 94% of cells were Pax6nuclearRx{thorn}, indicative of retinal progenitors. At prolonged induction, they expressed photoreceptor markers (Nrl, rhodopsin and its kinase) and showed significant responsiveness to excitatory glutamate. CONCLUSIONS. Primary human PDLSC could be directed to retinal progenitors with competence for photoreceptor differentiation. Human neural crest-derived PDL is readily accessible and can be an ample autologous source of undifferentiated cells for retinal cell regeneration.

Original languageEnglish (US)
Pages (from-to)3965-3974
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume54
Issue number6
DOIs
StatePublished - 2013

Keywords

  • Neurogenesis
  • Periodontal ligament-derived stem cells
  • Photoreceptors
  • Retinal progenitors

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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