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Gene Expression Profiling of Human Neural Progenitor Cells Following the Serum-Induced Astrocyte Differentiation

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Abstract

Neural stem cells (NSC) with self-renewal and multipotent properties could provide an ideal cell source for transplantation to treat spinal cord injury, stroke, and neurodegenerative diseases. However, the majority of transplanted NSC and neural progenitor cells (NPC) differentiate into astrocytes in vivo under pathological environments in the central nervous system, which potentially cause reactive gliosis. Because the serum is a potent inducer of astrocyte differentiation of rodent NPC in culture, we studied the effect of the serum on gene expression profile of cultured human NPC to identify the gene signature of astrocyte differentiation of human NPC. Human NPC spheres maintained in the serum-free culture medium were exposed to 10% fetal bovine serum (FBS) for 72 h, and processed for analyzing on a Whole Human Genome Microarray of 41,000 genes, and the microarray data were validated by real-time RT-PCR. The serum elevated the levels of expression of 45 genes, including ID1, ID2, ID3, CTGF, TGFA, METRN, GFAP, CRYAB and CSPG3, whereas it reduced the expression of 23 genes, such as DLL1, DLL3, PDGFRA, SOX4, CSPG4, GAS1 and HES5. Thus, the serum-induced astrocyte differentiation of human NPC is characterized by a counteraction of ID family genes on Delta family genes. Coimmunoprecipitation analysis identified ID1 as a direct binding partner of a proneural basic helix-loop-helix (bHLH) transcription factor MASH1. Luciferase assay indicated that activation of the DLL1 promoter by MASH1 was counteracted by ID1. Bone morphogenetic protein 4 (BMP4) elevated the levels of ID1 and GFAP expression in NPC under the serum-free culture conditions. Because the serum contains BMP4, these results suggest that the serum factor(s), most probably BMP4, induces astrocyte differentiation by upregulating the expression of ID family genes that repress the proneural bHLH protein-mediated Delta expression in human NPC.

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Abbreviations

NSC:

Neural stem cells

NPC:

Neural progenitor cells

CNS:

Central nervous system

BBB:

Blood–brain barrier

bHLH:

Basic helix-loop-helix

FBS:

Fetal bovine serum

EGF:

Epidermal growth factor

bFGF:

Basic fibroblast growth factor

LIF:

Leukemia inhibitory factor

TGF:

Transforming growth factor

RT-PCR:

Reverse transcription-polymerase chain reaction

DAVID:

Database for annotation visualization and integrated discovery

GO:

Gene Ontology

GFAP:

Glial fibrillary acidic protein

BMP4:

Bone morphogenetic protein 4

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Acknowledgments

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).

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Authors and Affiliations

  1. Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan

    Shinya Obayashi, Hiroko Tabunoki & Jun-ichi Satoh

  2. Division of Neurology, Department of Medicine, University of British Columbia Hospital, University of British Columbia, Vancouver, BC, Canada

    Seung U. Kim

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  1. Shinya Obayashi
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  2. Hiroko Tabunoki
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  3. Seung U. Kim
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  4. Jun-ichi Satoh
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Correspondence to Jun-ichi Satoh.

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Obayashi, S., Tabunoki, H., Kim, S.U. et al. Gene Expression Profiling of Human Neural Progenitor Cells Following the Serum-Induced Astrocyte Differentiation. Cell Mol Neurobiol 29, 423–438 (2009). https://doi.org/10.1007/s10571-008-9338-2

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