Cellular and Molecular Neurobiology

, Volume 30, Issue 3, pp 415–426 | Cite as

Stable Expression of Neurogenin 1 Induces LGR5, a Novel Stem Cell Marker, in an Immortalized Human Neural Stem Cell Line HB1.F3

  • Jun-ichi SatohEmail author
  • Shinya Obayashi
  • Hiroko Tabunoki
  • Taeko Wakana
  • Seung U. Kim
Original Research


Neural stem cells (NSC) with self-renewal and multipotent properties serve as an ideal cell source for transplantation to treat spinal cord injury, stroke, and neurodegenerative diseases. To efficiently induce neuronal lineage cells from NSC for neuron replacement therapy, we should clarify the intrinsic genetic programs involved in a time- and place-specific regulation of human NSC differentiation. Recently, we established an immortalized human NSC clone HB1.F3 to provide an unlimited NSC source applicable to genetic manipulation for cell-based therapy. To investigate a role of neurogenin 1 (Ngn1), a proneural basic helix-loop-helix (bHLH) transcription factor, in human NSC differentiation, we established a clone derived from F3 stably overexpressing Ngn1. Genome-wide gene expression profiling identified 250 upregulated genes and 338 downregulated genes in Ngn1-overexpressing F3 cells (F3-Ngn1) versus wild-type F3 cells (F3-WT). Notably, leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5), a novel stem cell marker, showed an 167-fold increase in F3-Ngn1, although transient overexpression of Ngn1 did not induce upregulation of LGR5, suggesting that LGR5 is not a direct transcriptional target of Ngn1. KeyMolnet, a bioinformatics tool for analyzing molecular relations on a comprehensive knowledgebase, suggests that the molecular network of differentially expressed genes involves the complex interaction of networks regulated by multiple transcription factors. Gene ontology (GO) terms of development and morphogenesis are enriched in upregulated genes, while those of extracellular matrix and adhesion are enriched in downregulated genes. These results suggest that stable expression of a single gene Ngn1 in F3 cells induces not simply neurogenic but multifunctional changes that potentially affect the differentiation of human NSC via a reorganization of complex gene regulatory networks.


HB1.F3 KeyMolnet LGR5 Microarray Neural stem cells Neurogenin 1 



Basic helix-loop-helix


Central nervous system


Database for annotation visualization and integrated discovery


Differentially expressed genes


Fetal bovine serum


Growth arrest-specific 2


Gene ontology


Hyaluronan synthase 2


Leucine-rich repeat-containing G protein-coupled receptor 5


Matrix metallopeptidase 9


Neurogenin 1


Neural progenitor cells


Neural stem cells


Open-reading frame


Robust multiarray average


Reverse transcription-polymerase chain reaction


Sonic hedgehog homolog


Wingless-type MMTV integration site family



This work was supported by a research grant to J-IS from the High-Tech Research Center Project, the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan (S0801043), and from Research on Intractable Diseases, the Ministry of Health, Labour and Welfare of Japan. The microarray data are available from Gene Expression Omnibus (GEO) under the accession number GSE18296.

Supplementary material

10571_2009_9466_MOESM1_ESM.ppt (280 kb)
Supplementary material 1 (PPT 280 kb) Supplementary Fig. 1. Upregulation of Ngn1 ORF expression in F3-Ngn1 cells. Genome-wide gene expression profiling of F3-WT and F3-Ngn1 was performed by using two sets of Human Gene 1.0 ST Array for each, followed by two comparisons composed of WT array-1 (F3-WT-1) versus Ngn1 array-1 (F3-Ngn1-1) and WT array-2 (F3-WT-2) versus Ngn1 array-2 (F3-Ngn1-2). The distribution pattern of individual probe-based signal intensities of NEUROG1 (Ngn1) on the array is shown. Among 26 probes spreading across the full-length Ngn1 gene, the position of the ORF is indicated by a purple box
10571_2009_9466_MOESM2_ESM.xls (61 kb)
Supplementary material 2 (XLS 61 kb)
10571_2009_9466_MOESM3_ESM.xls (73 kb)
Supplementary material 3 (XLS 73 kb)


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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Jun-ichi Satoh
    • 1
    Email author
  • Shinya Obayashi
    • 1
  • Hiroko Tabunoki
    • 1
  • Taeko Wakana
    • 1
  • Seung U. Kim
    • 2
    • 3
  1. 1.Department of Bioinformatics and Molecular NeuropathologyMeiji Pharmaceutical UniversityTokyoJapan
  2. 2.Division of Neurology, Department of MedicineUniversity of British Columbia Hospital, University of British ColumbiaVancouverCanada
  3. 3.Medical Research InstituteChung-Ang University College of MedicineSeoulKorea

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