Molecular and Cellular Biochemistry

, Volume 291, Issue 1–2, pp 1–10

Pluripotency potential of human adipose-derived stem cells marked with exogenous green fluorescent protein

  • Yunfeng Lin
  • Lei Liu
  • Zhiyong Li
  • Ju Qiao
  • Ling Wu
  • Wei Tang
  • Xiaohui Zheng
  • Xizhe Chen
  • Zhengbin Yan
  • Weidong Tian
Article
  • 210 Downloads

Abstract

Musculoskeletal tissues regeneration requires rapid expansion of seeding cells both in vitro and in vivo while maintaining their multilineage differentiation ability. Human adipose-derived stem cells (ASCs) are considered to contain multipotent mesenchymal stem cells. Monolayer cultures of human ASCs were isolated from human lipoaspirates and passaged 3 times and then infected with replication-incompetent adenoviral vectors carrying green fluorescent protein (Ad/GFP) genes. Then, Ad/GFP infected human ASCs were transferred to osteogenic, chondrogenic, adipogenic, and myogenic medium. The morphological characterization of induced cells was observed using phase-contrast microscopy and fluorescence microscopy. The expression of marker proteins or genes was measured by immunocytochemical and RT-PCR analysis. Osteopontin (OPN), and osteocalcin (OCN) were positive in osteogenic lineages, aggrecan and SOX9 were positive in chondrogenic ones, peroxisome proliferator-activated receptor (PPAR-γ2) and lipoprotein lipase (LPL) were positive in adipogenic ones, and myogenin and myod1 was positive in myogenic ones. At the same time, the results of fluorescence microscopic imaging proved that the high level of GFP expression during ASCs differentiation maintained stable nearly 2 months. So the exogenous GFP and multilineage potential of human ASCs had no severe influences on each other. Since the human ASCs can be easily obtained and abundant, it is proposed that they may be promising candidate cells for further studies on tissue engineering. Imaging with expression of GFP facilitates the research on ASCs physiological behavior and application in tissue engineering during differentiation both in vitro and in vivo.

Keywords

human adipose-derived stem cells green fluorescent protein pluripotency 

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Yunfeng Lin
    • 1
    • 3
  • Lei Liu
    • 1
  • Zhiyong Li
    • 1
  • Ju Qiao
    • 2
  • Ling Wu
    • 1
    • 3
  • Wei Tang
    • 1
  • Xiaohui Zheng
    • 1
  • Xizhe Chen
    • 1
  • Zhengbin Yan
    • 2
  • Weidong Tian
    • 1
    • 3
  1. 1.Department of Oral and Maxillofacial Surgery, West China College of StomatologySichuan UniversityChengduP.R. China
  2. 2.Department of Orthodontics, West China College of StomatologySichuan UniversityChengduP.R. China
  3. 3.Key Laboratory of Oral Biomedical EngineeringMinistry of Education. Sichuan UniversityChengduP.R. China

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