title: Single-cell time-of-flight secondary ion mass spectrometry reveals that human breast cancer stem cells have significantly lower content of palmitoleic acid compared to their counterpart non-stem cancer cells.
abstract: Lipids comprise the primary component of cell membranes. Imaging mass spectrometry is increasingly being used to visualize membranous lipids in clinical specimens, and it has revealed that abnormal lipid metabolism is related to the development of diseases. To characterize cell populations which are rare and sparsely localized in tissues, we conducted time-of-flight secondary ion mass spectrometry (TOF-SIMS) analyses of individual cells sorted by fluorescence activated cell sorting (FACS) and applied the method to analyze breast cancer stem cells (CSCs). TOF-SIMS analyses visualized phosphoric acids and four fatty acid (FA) species in the sorted CD45(-)/CD44(+)/CD24(-) CSCs, and these ions are suspected to have originated from membranous phospholipids as they were uniformly detected from the locus where the cells attached. Integrated ion intensity of palmitoleic acids [FA(16:1)] normalized by phosphoric acid signals were decreased significantly in CSCs as compared to that of CD45(-)/CD44(-)/CD24(+) non-stem cancer cells (NSCCs). This finding was supported by liquid chromatography coupled electrospray ionization-tandem mass spectrometry analysis, which revealed phosphatidylcholine (PC)(16:0/16:1) to be less abundant and PC(16:0/16:0) to be more abundant in CSCs as compared to NSCCs. Therefore, our novel method successfully provided lipid composition analysis of individual cells classified by the expression of a complex combination of cell-surface markers. The lipid compositions of CSCs originating from the heterogeneous cellular populations of clinical specimens were successfully characterized by this method.
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Michihiko
authors.initials: M
authors.lastname: Waki
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan; Department of Surgery I, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka, Japan.
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Ide
authors.affiliation: ULVAC-PHI, Inc., 370, Enzo, Chigasaki, Kanagawa, Japan.
authors.forename: Itsuko
authors.initials: I
authors.lastname: Ishizaki
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan; Oncology Center, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Nagata
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Noritaka
authors.initials: N
authors.lastname: Masaki
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Sugiyama
authors.affiliation: Department of Tumor Pathology, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka, Japan.
authors.forename: Nobuya
authors.initials: N
authors.lastname: Kurabe
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Dan
authors.initials: D
authors.lastname: Nicolaescu
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Fumiyoshi
authors.initials: F
authors.lastname: Yamazaki
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Hayasaka
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Ikegami
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kondo
authors.affiliation: Equipment Center, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Shibata
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Hiraide
authors.affiliation: Department of Surgery I, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Taki
authors.affiliation: Department of Surgery I, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ogura
authors.affiliation: Department of Surgery I, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka, Japan.
authors.forename: Norihiko
authors.initials: N
authors.lastname: Shiiya
authors.affiliation: ULVAC-PHI, Inc., 370, Enzo, Chigasaki, Kanagawa, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Sanada
authors.affiliation: Department of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1, Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan. Electronic address: setou@hama-med.ac.jp.
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Setou
mesh_terms: D000328:Adult; D001943:Breast Neoplasms / Q000737:chemistry* / Q000473:pathology; D051927:CD24 Antigen / Q000378:metabolism; D002853:Chromatography, Liquid; D005229:Fatty Acids, Monounsaturated / Q000032:analysis*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000737:chemistry* / Q000473:pathology; D010713:Phosphatidylcholines / Q000032:analysis; D059010:Single-Cell Analysis / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D051927:CD24 Antigen; D005229:Fatty Acids, Monounsaturated; D018960:Hyaluronan Receptors; D010713:Phosphatidylcholines; C008757:palmitoleic acid
keywords: Breast cancer; Cancer stem cell; Liquid chromatography coupled electrospray ionization–tandem mass spectrometry; Palmitoleic acid; Time-of-flight secondary ion mass spectrometry