title: [Isolation and in vitro characterization of CD133(+) side population cells from laryngeal cancer cell line].
abstract: OBJECTIVE
  To investigate an approach enriching cancer stem cells (CSCs) more effectively from laryngeal cancer cell line.
  
  
  METHODS
  CD133(+)SP and CD133(-)SP subpopulation was detected and isolated from Hep-2 cell line using Hoechst33342 dye and phycoerythrin (PE)-conjugated CD133 monoclonal antibody assisted by fluorescence activated cell sorting technology. Sorted CD133(+)SP and CD133(-)SP cells were compared in CSCs-related assays including proliferation, differentiation, spheroid formation and drug sensitivity.
  
  
  RESULTS
  CD133(+)SP cells accounted for a very small fraction of (0.30 ± 0.12)% in Hep-2 cell line, far less than the proportion of CD133(+) subgroup and side population subgroup, which were (3.15 ± 0.83)% and (17.1 ± 2.0)% respectively. Intriguingly, CD133(+)SP cells proliferated much faster than CD133(-)SP cells in RPMI1640 and gave rise to CD133(-)SP cells and other heterogeneous cells that formed the bulk of the tumor. In contrast, CD133(-)SP cells were not able to differentiate into CD133(+)SP cells. In serum-free medium CD133(+)SP cells grew as spherical clusters and remained floating. In addition, CD133(+)SP cells manifested the marked resistance to chemotherapy than CD133(-)SP cells.
  
  
  CONCLUSIONS
  Compared with CD133(-)SP cells, CD133(+)SP subpopulation exhibited extraordinary cancer stem-like properties, were enriched for cancer stem cells more effectively and might serve as an ideal putative candidate for CSCs research in laryngeal cancer.
authors.affiliation: Department of Otorhinolaryngology Head and Neck Surgery, Fudan University Affiliated Eye Ear Nose and Throat Hospital, Shanghai 200031, China.
authors.forename: Chun-ping
authors.initials: CP
authors.lastname: Wu
authors.forename: Liang
authors.initials: L
authors.lastname: Zhou
authors.forename: Ming
authors.initials: M
authors.lastname: Xie
authors.forename: Lei
authors.initials: L
authors.lastname: Tao
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Tian
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002469:Cell Separation; D006023:Glycoproteins; D006801:Humans; D007822:Laryngeal Neoplasms / Q000473:pathology; D014411:Neoplastic Stem Cells / Q000166:cytology*; D010455:Peptides; D058985:Side-Population Cells / Q000166:cytology*
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides