title: [Isolation and characterization of human breast tumor stem cells].
abstract: OBJECTIVE
  To isolate and culture human breast cancer stem cells,  and characterize their biological properties in vitro.
  
  
  METHODS
  Fresh tumor tissues of breast cancer patients were collected from surgical rooms. The tissue samples were mechanically sheared and enzyme-digested to get single cell suspensions. Cancer cell surface antigens were analyzed by flow cytometry,  and cell growth curve was established by MTT assay;  Real-time quantitative PCR (qPCR) was employed to detect the expressions of stem cell-specific genes Sox2 and Nanog in the cancer cells;  Immunofluorescence staining was performed to examine  the expressions of breast cancer-specific proteins Bcl-2 and progesterone receptor (PR).
  
  
  RESULTS
  Human breast cancer stem cells grew in the form of spheres. These stem cells had a CD44(+);/CD24(-); phenotype as characterized by flow cytometry,  expressed high levels of Sox2 and Nanog genes,  and were positive for Bcl-2 and PR.
  
  
  CONCLUSIONS
  Human breast tumors contain CD44(+);/CD24(-); cancer stem cells which are capable of self-renewal and proliferation,  and can be long-term cultured and expanded in vitro.
authors.affiliation: Institute of Stem Cells, General Hospital, Ministry of Education, Ningxia Medical University, Yinchuan 750004, China. wanglinbin007@126.com
authors.forename: Li-bin
authors.initials: LB
authors.lastname: Wang
authors.forename: Ya-qin
authors.initials: YQ
authors.lastname: He
authors.forename: Li-gang
authors.initials: LG
authors.lastname: Wu
authors.forename: Dong-mei
authors.initials: DM
authors.lastname: Chen
authors.forename: Heng
authors.initials: H
authors.lastname: Fan
authors.forename: Wei
authors.initials: W
authors.lastname: Jia
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism / Q000503:physiopathology*; D051927:CD24 Antigen / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000166:cytology* / Q000378:metabolism
chemical_list: D051927:CD24 Antigen; D018960:Hyaluronan Receptors