title: The application of an in situ karyotyping technique for mesenchymal stromal cells: a validation and comparison study with classical G-banding.
abstract: The cytogenetic analysis of mesenchymal stromal cells (MSCs) is essential for verifying the safety and stability of MSCs. An in situ technique, which uses cells grown on coverslips for karyotyping and minimizes cell manipulation, is the standard protocol for the chromosome analysis of amniotic fluids. Therefore, we applied the in situ karyotyping technique in MSCs and compared the quality of metaphases and karyotyping results with classical G-banding and chromosomal abnormalities with fluorescence in situ hybridization (FISH). Human adipose- and umbilical cord-derived MSC cell lines (American Type Culture Collection PCS-500-011, PCS-500-010) were used for evaluation. The quality of metaphases was assessed by analyzing the chromosome numbers in each metaphase, the overlaps of chromosomes and the mean length of chromosome 1. FISH was performed in the interphase nuclei of MSCs for 6q, 7q and 17q abnormalities and for the enumeration of chromosomes via oligo-FISH in adipose-derived MSCs. The number of chromosomes in each metaphase was more variable in classical G-banding. The overlap of chromosomes and the mean length of chromosome 1 as observed via in situ karyotyping were comparable to those of classical G-banding (P=0.218 and 0.674, respectively). Classical G-banding and in situ karyotyping by two personnel showed normal karyotypes for both cell lines in five passages. No numerical or structural chromosomal abnormalities were found by the interphase-FISH. In situ karyotyping showed equivalent karyotype results, and the quality of the metaphases was not inferior to classical G-banding. Thus, in situ karyotyping with minimized cell manipulation and the use of less cells would be useful for karyotyping MSCs.
authors.affiliation: 1] Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Korea [2] Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Sang Mee
authors.initials: SM
authors.lastname: Hwang
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Cha-Ja
authors.initials: CJ
authors.lastname: See
authors.affiliation: Department of Laboratory Medicine, Korea University Hospital, Seoul, Korea.
authors.forename: Jungeun
authors.initials: J
authors.lastname: Choi
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Seon Young
authors.initials: SY
authors.lastname: Kim
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Qute
authors.initials: Q
authors.lastname: Choi
authors.affiliation: Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Jung Ah
authors.initials: JA
authors.lastname: Kim
authors.affiliation: Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Jiseok
authors.initials: J
authors.lastname: Kwon
authors.affiliation: Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Si Nae
authors.initials: SN
authors.lastname: Park
authors.affiliation: Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Kyongok
authors.initials: K
authors.lastname: Im
authors.affiliation: Catholic High-Performance Cell Therapy Center, Department of Cellular Medicine, Research Center for Stem Cell Therapeutics Evaluation, Catholic University of Korea, Seoul, Korea.
authors.forename: Il-Hoan
authors.initials: IH
authors.lastname: Oh
authors.affiliation: 1] Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea [2] Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Dong Soon
authors.initials: DS
authors.lastname: Lee
mesh_terms: D001399:Azure Stains; D002871:Chromosome Banding / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007621:Karyotyping / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology*
chemical_list: D001399:Azure Stains