title: [Enrichment and function research of large cell lung cancer stem cell-like cells].
abstract: OBJECTIVE
  There are no universal method to recognize and screen for lung cancer stem cell markers and indicators. Commonly used methods are flow Cytometry and learning from other cancer stem cell sorting tags to sort lung cancer stem cells. But this method has low specificity screening, the workload is huge. In this study, Serum-free suspension culture was used to enrich lung cancer stem cells, and explore method for lung cancer stem cell screening.
  
  
  METHODS
  Human large lung cancer cell line-L9981 was cultured in serum-free and growth factors added medium, and spheres were obtained. Then the morphological differences of sphere cells and adherent L9981 cells cultured in serum-containing mediums are observed. Cell proliferation was analyzed by Vi-cell viability analyzer; invasion ability was tested by transwell assay; and in vivo tumorigenicity of the two groups of cells was studied in nude mouse.
  
  
  RESULTS
  Compared with adherent L9981 cells cultured in serum-containing mediums, cells cultured in serum-free medium display sphere appearance. Doubling time of adherent cells and sphere cells are (56.05±1.95) h and (33.00±1.44) h respectively; Spheroid cells had higher invasion and tumorigenicity ability, 5 times and 20 times respectively, than adherent cells.
  
  
  CONCLUSIONS
  Suspension cultured L9981 in Serum-free medium could form spheroid populations. Cells in spheres had higher ability of invasion and Tumorigenicity than adherent L9981 cells. These results indicated spheroid L9981 cells contained enriched lung cancer stem cells, and Serum-free suspension culture can be a candidate method for enriching lung cancer stem cell.
authors.affiliation: Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors.forename: Wenke
authors.initials: W
authors.lastname: Yue
authors.forename: Feng
authors.initials: F
authors.lastname: Jiao
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.forename: Jiacong
authors.initials: J
authors.lastname: You
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000818:Animals; D018287:Carcinoma, Large Cell / Q000473:pathology*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002469:Cell Separation; D002471:Cell Transformation, Neoplastic; D016895:Culture Media, Serum-Free; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000473:pathology*
chemical_list: D016895:Culture Media, Serum-Free