Role of transcription factors in motoneuron differentiation of adult human olfactory neuroepithelial-derived progenitors.
Neurosphereforming cell (NSFC) lines have been established from cultures of human adult olfactory neuroepithelium. Few of these cells ever express mature neuronal or glial markers in minimal essential medium supplemented with 10% fetal bovine serum or defined medium. However, these neural progenitors have the potential to differentiate along glial or neuronal lineages. To evaluate the potential of NSFCs to form motoneurons, transcription factors Olig2, Ngn2, and HB9 were introduced into NSFCs to determine if their expression is sufficient for motoneuron specification and differentiation, as has been shown in the early development of the avian and murine central nervous systems in vivo. NSFCs transfected with Olig2, Ngn2, and HB9 alone exhibited no phenotypic lineage restriction. In contrast, simultaneous transfection of Ngn2 and HB9 cDNA increased the expression of Isl1/2, a motoneuron marker, when the cells were maintained in medium supplemented with retinoic acid, forskolin, and sonic hedgehog. Furthermore, a population of Olig2-expressing NSFCs also expressed Ngn2. Cotransfection of NSFCs with Olig2 and HB9, but not Olig2 and Ngn2, increased Isl1/2 expression. Coculture of NSFCs trans-fected with Ngn2-HB92 or Olig2 and HB9 with purified chicken skeletal muscle demonstrated frequent contacts that resembled neuromuscular junctions. These studies demonstrate that transcription factors governing the early development of chick and mouse motoneuron formation are able to drive human adult olfactory neuroepithelial progenitors to differentiate into motoneurons in vitro. Our long-term goal is to develop cell populations for future studies of the therapeutic utility of these olfactory-derived NSFCs for autologous cell replacement strategies for central nervous system trauma and neurodegenerative diseases.
Zhang
Xiaodong
X
Department of Anatomical Sciences and Neurobiology, University of Louisville, School of Medicine, KY 40202, USA.
Cai
Jun
J
Klueber
Kathleen M
KM
Guo
Zhanfang
Z
Lu
Chengliang
C
Winstead
Welby I
WI
Qiu
Mengsheng
M
Roisen
Fred J
FJ
D000328:Adult; D000369:Aged, 80 and over; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism / Q000502:physiology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D002645:Chickens; D018920:Coculture Techniques; D005260:Female; D018398:Homeodomain Proteins / Q000378:metabolism / Q000502:physiology; D006801:Humans; D009046:Motor Neurons / Q000502:physiology*; D018482:Muscle, Skeletal / Q000166:cytology; D009419:Nerve Tissue Proteins / Q000378:metabolism / Q000502:physiology*; D009832:Olfactory Nerve / Q000166:cytology*; D000076104:Oligodendrocyte Transcription Factor 2; D010641:Phenotype; D013234:Stem Cells / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*; D014162:Transfection
D051792:Basic Helix-Loop-Helix Proteins; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D000076104:Oligodendrocyte Transcription Factor 2; D014157:Transcription Factors; C491183:NEUROG2 protein, human; C102938:Neurog2 protein, mouse; C405708:OLIG2 protein, human; C512773:Olig2 protein, mouse; C084059:Hb9 protein, mouse