Skip to main content
placeholder image

Directing Human Induced Pluripotent Stem Cells into a Neurosensory Lineage for Auditory Neuron Replacement

Journal Article


Download full-text (Open Access)

Abstract


  • Emerging therapies for sensorineural hearing loss include replacing damaged auditory neurons (ANs) using stem cells. Ultimately, it is important that these replacement cells can be patient-matched to avoid immunorejection. As human induced pluripotent stem cells (hiPSCs) can be obtained directly from the patient, they offer an opportunity to generate patient-matched neurons for transplantation. Here, we used an established neural induction protocol to differentiate two hiPSC lines (iPS1 and iPS2) and one human embryonic stem cell line (hESC; H9) toward a neurosensory lineage in vitro. Immunocytochemistry and qRT-PCR were used to analyze the expression of key markers involved in AN development at defined time points of differentiation. The hiPSC- and hESCderived neurosensory progenitors expressed the dorsal hindbrain marker (PAX7), otic placodal marker (PAX2), proneurosensory marker (SOX2), ganglion neuronal markers (NEUROD1, BRN3A, ISLET1, ├čIII-tubulin, Neurofilament kDa 160), and sensory AN markers (GATA3 and VGLUT1) over the time course examined. The hiPSC- and hESC-derived neurosensory progenitors had the highest expression levels of the sensory neural markers at 35 days in vitro. Furthermore, the neurons generated from this assay were found to be electrically active. While all cell lines analyzed produced functional neurosensory-like progenitors, variabilities in the levels of marker expression were observed between hiPSC lines and within samples of the same cell line, when compared with the hESC controls. Overall, these findings indicate that this neural assay was capable of differentiating hiPSCs toward a neurosensory lineage but emphasize the need for improving the consistency in the differentiation of hiPSCs into the required lineages.

Authors


  •   Gunewardene, Niliksha (external author)
  •   Van Bergen, Nicole (external author)
  •   Crombie, Duncan E. (external author)
  •   Needham, Karina (external author)
  •   Dottori, Mirella
  •   Nayagam, Bryony A. (external author)

Publication Date


  • 2014

Citation


  • Gunewardene, N., Van Bergen, N., Crombie, D., Needham, K., Dottori, M. & Nayagam, B. A. (2014). Directing Human Induced Pluripotent Stem Cells into a Neurosensory Lineage for Auditory Neuron Replacement. BioResearch Open Access, 3 (4), 162-175.

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=2188&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1162

Number Of Pages


  • 13

Start Page


  • 162

End Page


  • 175

Volume


  • 3

Issue


  • 4

Place Of Publication


  • United States

Abstract


  • Emerging therapies for sensorineural hearing loss include replacing damaged auditory neurons (ANs) using stem cells. Ultimately, it is important that these replacement cells can be patient-matched to avoid immunorejection. As human induced pluripotent stem cells (hiPSCs) can be obtained directly from the patient, they offer an opportunity to generate patient-matched neurons for transplantation. Here, we used an established neural induction protocol to differentiate two hiPSC lines (iPS1 and iPS2) and one human embryonic stem cell line (hESC; H9) toward a neurosensory lineage in vitro. Immunocytochemistry and qRT-PCR were used to analyze the expression of key markers involved in AN development at defined time points of differentiation. The hiPSC- and hESCderived neurosensory progenitors expressed the dorsal hindbrain marker (PAX7), otic placodal marker (PAX2), proneurosensory marker (SOX2), ganglion neuronal markers (NEUROD1, BRN3A, ISLET1, ├čIII-tubulin, Neurofilament kDa 160), and sensory AN markers (GATA3 and VGLUT1) over the time course examined. The hiPSC- and hESC-derived neurosensory progenitors had the highest expression levels of the sensory neural markers at 35 days in vitro. Furthermore, the neurons generated from this assay were found to be electrically active. While all cell lines analyzed produced functional neurosensory-like progenitors, variabilities in the levels of marker expression were observed between hiPSC lines and within samples of the same cell line, when compared with the hESC controls. Overall, these findings indicate that this neural assay was capable of differentiating hiPSCs toward a neurosensory lineage but emphasize the need for improving the consistency in the differentiation of hiPSCs into the required lineages.

Authors


  •   Gunewardene, Niliksha (external author)
  •   Van Bergen, Nicole (external author)
  •   Crombie, Duncan E. (external author)
  •   Needham, Karina (external author)
  •   Dottori, Mirella
  •   Nayagam, Bryony A. (external author)

Publication Date


  • 2014

Citation


  • Gunewardene, N., Van Bergen, N., Crombie, D., Needham, K., Dottori, M. & Nayagam, B. A. (2014). Directing Human Induced Pluripotent Stem Cells into a Neurosensory Lineage for Auditory Neuron Replacement. BioResearch Open Access, 3 (4), 162-175.

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=2188&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1162

Number Of Pages


  • 13

Start Page


  • 162

End Page


  • 175

Volume


  • 3

Issue


  • 4

Place Of Publication


  • United States