Human dental stem cell derived transgene-free iPSCs generate functional neurons via embryoid body-mediated and direct induction methods

Ikbale El Ayachi, Jun Zhang, Xiao Ying Zou, Dong Li, Zongdong Yu, Wei Wei, Kristen M.S. O'Connell, George Huang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Induced pluripotent stem cells (iPSCs) give rise to neural stem/progenitor cells, serving as a good source for neural regeneration. Here, we established transgene-free (TF) iPSCs from dental stem cells (DSCs) and determined their capacity to differentiate into functional neurons in vitro. Generated TF iPSCs from stem cells of apical papilla and dental pulp stem cells underwent two methods—embryoid body-mediated and direct induction, to guide TF-DSC iPSCs along with H9 or H9 Syn-GFP (human embryonic stem cells) into functional neurons in vitro. Using the embryoid body-mediated method, early stage neural markers PAX6, SOX1, and nestin were detected by immunocytofluorescence or reverse transcription-real time polymerase chain reaction (RT-qPCR). At late stage of neural induction measured at Weeks 7 and 9, the expression levels of neuron-specific markers Nav1.6, Kv1.4, Kv4.2, synapsin, SNAP25, PSD95, GAD67, GAP43, and NSE varied between stem cells of apical papilla iPSCs and H9. For direct induction method, iPSCs were directly induced into neural stem/progenitor cells and guided to become neuron-like cells. The direct method, while simpler, showed cell detachment and death during the differentiation process. At early stage, PAX6, SOX1 and nestin were detected. At late stage of differentiation, all five genes tested, nestin, βIII-tubulin, neurofilament medium chain, GFAP, and Nav, were positive in many cells in cultures. Both differentiation methods led to neuron-like cells in cultures exhibiting sodium and potassium currents, action potential, or spontaneous excitatory postsynaptic potential. Thus, TF-DSC iPSCs are capable of undergoing guided neurogenic differentiation into functional neurons in vitro, thereby may serve as a cell source for neural regeneration.

Original languageEnglish (US)
Pages (from-to)e1836-e1851
JournalJournal of Tissue Engineering and Regenerative Medicine
Volume12
Issue number4
DOIs
StatePublished - Apr 1 2018

Fingerprint

Embryoid Bodies
Induced Pluripotent Stem Cells
Stem cells
Transgenes
Neurons
Tooth
Stem Cells
Nestin
Neural Stem Cells
Regeneration
Cell Culture Techniques
Miniature Postsynaptic Potentials
Synapsins
Dental Pulp
Intermediate Filaments
Excitatory Postsynaptic Potentials
Tubulin
Action Potentials
Reverse Transcription
Real-Time Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Medicine (miscellaneous)
  • Biomaterials
  • Biomedical Engineering

Cite this

Human dental stem cell derived transgene-free iPSCs generate functional neurons via embryoid body-mediated and direct induction methods. / El Ayachi, Ikbale; Zhang, Jun; Zou, Xiao Ying; Li, Dong; Yu, Zongdong; Wei, Wei; O'Connell, Kristen M.S.; Huang, George.

In: Journal of Tissue Engineering and Regenerative Medicine, Vol. 12, No. 4, 01.04.2018, p. e1836-e1851.

Research output: Contribution to journalArticle

El Ayachi, Ikbale ; Zhang, Jun ; Zou, Xiao Ying ; Li, Dong ; Yu, Zongdong ; Wei, Wei ; O'Connell, Kristen M.S. ; Huang, George. / Human dental stem cell derived transgene-free iPSCs generate functional neurons via embryoid body-mediated and direct induction methods. In: Journal of Tissue Engineering and Regenerative Medicine. 2018 ; Vol. 12, No. 4. pp. e1836-e1851.
@article{3531d953120140b48676dfe5eaa12997,
title = "Human dental stem cell derived transgene-free iPSCs generate functional neurons via embryoid body-mediated and direct induction methods",
abstract = "Induced pluripotent stem cells (iPSCs) give rise to neural stem/progenitor cells, serving as a good source for neural regeneration. Here, we established transgene-free (TF) iPSCs from dental stem cells (DSCs) and determined their capacity to differentiate into functional neurons in vitro. Generated TF iPSCs from stem cells of apical papilla and dental pulp stem cells underwent two methods—embryoid body-mediated and direct induction, to guide TF-DSC iPSCs along with H9 or H9 Syn-GFP (human embryonic stem cells) into functional neurons in vitro. Using the embryoid body-mediated method, early stage neural markers PAX6, SOX1, and nestin were detected by immunocytofluorescence or reverse transcription-real time polymerase chain reaction (RT-qPCR). At late stage of neural induction measured at Weeks 7 and 9, the expression levels of neuron-specific markers Nav1.6, Kv1.4, Kv4.2, synapsin, SNAP25, PSD95, GAD67, GAP43, and NSE varied between stem cells of apical papilla iPSCs and H9. For direct induction method, iPSCs were directly induced into neural stem/progenitor cells and guided to become neuron-like cells. The direct method, while simpler, showed cell detachment and death during the differentiation process. At early stage, PAX6, SOX1 and nestin were detected. At late stage of differentiation, all five genes tested, nestin, βIII-tubulin, neurofilament medium chain, GFAP, and Nav, were positive in many cells in cultures. Both differentiation methods led to neuron-like cells in cultures exhibiting sodium and potassium currents, action potential, or spontaneous excitatory postsynaptic potential. Thus, TF-DSC iPSCs are capable of undergoing guided neurogenic differentiation into functional neurons in vitro, thereby may serve as a cell source for neural regeneration.",
author = "{El Ayachi}, Ikbale and Jun Zhang and Zou, {Xiao Ying} and Dong Li and Zongdong Yu and Wei Wei and O'Connell, {Kristen M.S.} and George Huang",
year = "2018",
month = "4",
day = "1",
doi = "10.1002/term.2615",
language = "English (US)",
volume = "12",
pages = "e1836--e1851",
journal = "Journal of Tissue Engineering and Regenerative Medicine",
issn = "1932-6254",
publisher = "John Wiley and Sons Ltd",
number = "4",

}

TY - JOUR

T1 - Human dental stem cell derived transgene-free iPSCs generate functional neurons via embryoid body-mediated and direct induction methods

AU - El Ayachi, Ikbale

AU - Zhang, Jun

AU - Zou, Xiao Ying

AU - Li, Dong

AU - Yu, Zongdong

AU - Wei, Wei

AU - O'Connell, Kristen M.S.

AU - Huang, George

PY - 2018/4/1

Y1 - 2018/4/1

N2 - Induced pluripotent stem cells (iPSCs) give rise to neural stem/progenitor cells, serving as a good source for neural regeneration. Here, we established transgene-free (TF) iPSCs from dental stem cells (DSCs) and determined their capacity to differentiate into functional neurons in vitro. Generated TF iPSCs from stem cells of apical papilla and dental pulp stem cells underwent two methods—embryoid body-mediated and direct induction, to guide TF-DSC iPSCs along with H9 or H9 Syn-GFP (human embryonic stem cells) into functional neurons in vitro. Using the embryoid body-mediated method, early stage neural markers PAX6, SOX1, and nestin were detected by immunocytofluorescence or reverse transcription-real time polymerase chain reaction (RT-qPCR). At late stage of neural induction measured at Weeks 7 and 9, the expression levels of neuron-specific markers Nav1.6, Kv1.4, Kv4.2, synapsin, SNAP25, PSD95, GAD67, GAP43, and NSE varied between stem cells of apical papilla iPSCs and H9. For direct induction method, iPSCs were directly induced into neural stem/progenitor cells and guided to become neuron-like cells. The direct method, while simpler, showed cell detachment and death during the differentiation process. At early stage, PAX6, SOX1 and nestin were detected. At late stage of differentiation, all five genes tested, nestin, βIII-tubulin, neurofilament medium chain, GFAP, and Nav, were positive in many cells in cultures. Both differentiation methods led to neuron-like cells in cultures exhibiting sodium and potassium currents, action potential, or spontaneous excitatory postsynaptic potential. Thus, TF-DSC iPSCs are capable of undergoing guided neurogenic differentiation into functional neurons in vitro, thereby may serve as a cell source for neural regeneration.

AB - Induced pluripotent stem cells (iPSCs) give rise to neural stem/progenitor cells, serving as a good source for neural regeneration. Here, we established transgene-free (TF) iPSCs from dental stem cells (DSCs) and determined their capacity to differentiate into functional neurons in vitro. Generated TF iPSCs from stem cells of apical papilla and dental pulp stem cells underwent two methods—embryoid body-mediated and direct induction, to guide TF-DSC iPSCs along with H9 or H9 Syn-GFP (human embryonic stem cells) into functional neurons in vitro. Using the embryoid body-mediated method, early stage neural markers PAX6, SOX1, and nestin were detected by immunocytofluorescence or reverse transcription-real time polymerase chain reaction (RT-qPCR). At late stage of neural induction measured at Weeks 7 and 9, the expression levels of neuron-specific markers Nav1.6, Kv1.4, Kv4.2, synapsin, SNAP25, PSD95, GAD67, GAP43, and NSE varied between stem cells of apical papilla iPSCs and H9. For direct induction method, iPSCs were directly induced into neural stem/progenitor cells and guided to become neuron-like cells. The direct method, while simpler, showed cell detachment and death during the differentiation process. At early stage, PAX6, SOX1 and nestin were detected. At late stage of differentiation, all five genes tested, nestin, βIII-tubulin, neurofilament medium chain, GFAP, and Nav, were positive in many cells in cultures. Both differentiation methods led to neuron-like cells in cultures exhibiting sodium and potassium currents, action potential, or spontaneous excitatory postsynaptic potential. Thus, TF-DSC iPSCs are capable of undergoing guided neurogenic differentiation into functional neurons in vitro, thereby may serve as a cell source for neural regeneration.

UR - http://www.scopus.com/inward/record.url?scp=85040722794&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85040722794&partnerID=8YFLogxK

U2 - 10.1002/term.2615

DO - 10.1002/term.2615

M3 - Article

VL - 12

SP - e1836-e1851

JO - Journal of Tissue Engineering and Regenerative Medicine

JF - Journal of Tissue Engineering and Regenerative Medicine

SN - 1932-6254

IS - 4

ER -