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A Novel Monoclonal Antibody Against Neuroepithelial and Ependymal Cells and Characteristics of Its Positive Cells in Neurospheres

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Abstract

There are still few useful cell membrane surface antigens suitable for identification and isolation of neural stem cells (NSCs). We generated a novel monoclonal antibody (mAb), designated as mAb against immature neural cell antigens (INCA mAb), which reacted with the areas around a lateral ventricle of a fetal cerebrum. INCA mAb specifically reacted with neuroepithelial cells in fetal cerebrums and ependymal cells in adult cerebrums. The recognition molecules were O-linked 40 and 42 kDa glycoproteins on the cell membrane surface (gp40 INCA and gp42 INCA). Based on expression pattern analysis of the recognition molecules in developing cerebrums, it was concluded that gp42 INCA was a stage-specific antigen expressed on undifferentiated neuroepithelial cells, while gp40 INCA was a cell lineage-specific antigen expressed at the stages of differentiation from neuroepithelial cells to ependymal cells. A flow cytometric analysis showed that fetal and young adult neurospheres were divided into INCA mAb− CD133 polyclonal antibody (pAb)−, INCA mAb+ CD133 pAb−, and INCA mAb+ CD133 pAb+ cell populations based on the reactivity against INCA mAb and CD133 pAb. The proportion of cells having the neurosphere formation capability in the INCA mAb+ CD133 pAb+ cell population was significantly larger than that of undivided neurospheres. Neurospheres formed by clonal expansion of INCA mAb+ CD133 pAb+ cells gave rise to neurons and glial cells. INCA mAb will be a useful immunological probe in the study of NSCs.

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Abbreviations

Ab:

Antibody

BSA:

Bovine serum albumin

CNE:

Cortical neuroepithelium

CP:

Caudate putamen

ECL:

Ependymal cell layer

EGF:

Epidermal growth factor

FACS:

Fluorescence-activated cell sorting

b-FGF:

Basic-fibroblast growth factor

GFAP:

Glial fibrillary acidic protein

O-Glycosidase:

End-α-N-acetylgalactosaminidase

LIF:

Leukemia inhibitory factor

LMS:

Lateral migratory stream

LV:

Lateral ventricle (s)

mAb:

Monoclonal antibody

NSC:

Neural stem cell

pAb:

Polyclonal antibody

PBS:

Phosphate-buffered saline

PNGase F:

Peptide-N-glycosidase F

PVDF:

Polyvinylidene difluoride

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SeNE:

Septal neuroepithelium

StNE:

Striatal neuroepithelium

SVZ:

Subventricular zone

VZ:

Ventricular zone

VZ/SZV:

VZ plus SVZ

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Acknowledgments

This study was partially supported by Ohu University research funding.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Molecular and Cellular Biology, Faculty of Pharmaceutical Sciences, Ohu University, Fukushima, 963-8611, Japan

    Masaharu Kotani

  2. Department of Health Chemistry, Faculty of Pharmaceutical Sciences, Ohu University, Fukushima, 963-8611, Japan

    Yasunori Sato & Akemichi Ueno

  3. Department of English Language Technology, Faculty of Pharmaceutical Sciences, Ohu University, Fukushima, 963-8611, Japan

    Toshinori Ito

  4. Laboratory for Pharmacotherapy and Experimental Neurology, Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, Kagawa, 769-2193, Japan

    Kouichi Itoh

  5. Department of Functional Morphology, Nihon University School of Medicine, Tokyo, 173-8610, Japan

    Masato Imada

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  1. Masaharu Kotani
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  2. Yasunori Sato
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  3. Akemichi Ueno
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  4. Toshinori Ito
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Correspondence to Masaharu Kotani.

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Kotani, M., Sato, Y., Ueno, A. et al. A Novel Monoclonal Antibody Against Neuroepithelial and Ependymal Cells and Characteristics of Its Positive Cells in Neurospheres. Cell Mol Neurobiol 36, 11–26 (2016). https://doi.org/10.1007/s10571-015-0216-4

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  • DOI: https://doi.org/10.1007/s10571-015-0216-4

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