title: High efficient isolation and systematic identification of human adipose-derived mesenchymal stem cells.
abstract: BACKGROUND
  Developing efficient methods to isolate and identify human adipose-derived mesenchymal stem cells (hADSCs) remains to be one of the major challenges in tissue engineering.
  
  
  METHODS
  We demonstrate here a method by isolating hADSCs from abdominal subcutaneous adipose tissue harvested during caesarian section. The hADSCs were isolated from human adipose tissue by collagenase digestion and adherence to flasks.
  
  
  RESULTS
  The yield reached around 1 × 10(6) hADSCs per gram adipose tissue. The following comprehensive identification and characterization illustrated pronounced features of mesenchymal stem cells (MSCs). The fibroblast-like hADSCs exhibited typical ultrastructure details for vigorous cell activities. Karyotype mapping showed normal human chromosome. With unique immunophenotypes they were positive for CD29, CD44, CD73, CD105 and CD166, but negative for CD31, CD34, CD45 and HLA-DR. The growth curve and cell cycle analysis revealed high capability for self-renewal and proliferation. Moreover, these cells could be functionally induced into adipocytes, osteoblasts, and endothelial cells in the presence of appropriate conditioned media.
  
  
  CONCLUSIONS
  The data presented here suggest that we have developed high efficient isolation and cultivation methods with a systematic strategy for identification and characterization of hADSCs. These techniques will be able to provide safe and stable seeding cells for research and clinical application.
authors.affiliation: Key Laboratory of Pathobiology, Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, China.
authors.forename: Xu-Fang
authors.initials: XF
authors.lastname: Yang
authors.forename: Xu
authors.initials: X
authors.lastname: He
authors.forename: Jian
authors.initials: J
authors.lastname: He
authors.forename: Li-Hong
authors.initials: LH
authors.lastname: Zhang
authors.forename: Xue-Jin
authors.initials: XJ
authors.lastname: Su
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Dong
authors.forename: Yun-Jian
authors.initials: YJ
authors.lastname: Xu
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Yu-Lin
authors.initials: YL
authors.lastname: Li
mesh_terms: D000328:Adult; D002453:Cell Cycle / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D017364:Collagenases; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D007621:Karyotyping; D059630:Mesenchymal Stem Cells / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission; D060888:Real-Time Polymerase Chain Reaction; D050151:Subcutaneous Fat / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D017364:Collagenases