title: A Novel Monoclonal Antibody Against Neuroepithelial and Ependymal Cells and Characteristics of Its Positive Cells in Neurospheres.
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.
authors.affiliation: Department of Molecular and Cellular Biology, Faculty of Pharmaceutical Sciences, Ohu University, Fukushima, 963-8611, Japan. m-kotani@pha.ohu-u.ac.jp.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Kotani
authors.affiliation: Department of Health Chemistry, Faculty of Pharmaceutical Sciences, Ohu University, Fukushima, 963-8611, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Sato
authors.affiliation: Department of Health Chemistry, Faculty of Pharmaceutical Sciences, Ohu University, Fukushima, 963-8611, Japan.
authors.forename: Akemichi
authors.initials: A
authors.lastname: Ueno
authors.affiliation: Department of English Language Technology, Faculty of Pharmaceutical Sciences, Ohu University, Fukushima, 963-8611, Japan.
authors.forename: Toshinori
authors.initials: T
authors.lastname: Ito
authors.affiliation: Laboratory for Pharmacotherapy and Experimental Neurology, Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, Kagawa, 769-2193, Japan.
authors.forename: Kouichi
authors.initials: K
authors.lastname: Itoh
authors.affiliation: Department of Functional Morphology, Nihon University School of Medicine, Tokyo, 173-8610, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Imada
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity / Q000276:immunology; D002469:Cell Separation; D054022:Cerebrum / Q000196:embryology; D004805:Ependyma / Q000378:metabolism*; D005260:Female; D005333:Fetus / Q000166:cytology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D008297:Male; D008819:Mice, Nude; D064231:Nestin / Q000378:metabolism; D046569:Neuroepithelial Cells / Q000378:metabolism*; D010766:Phosphorylation; D011916:Rats, Inbred F344; D018874:Spheroids, Cellular / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D005904:Glial Fibrillary Acidic Protein; D006657:Histones; D064231:Nestin
keywords: Cell membrane surface antigen; Ependymal cell; Monoclonal antibody; Neural stem cell; Neuroepithelial cell; Neurosphere