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Molecular and Cellular Biochemistry

, Volume 285, Issue 1–2, pp 69–78 | Cite as

Multilineage differentiation of adipose-derived stromal cells from GFP transgenic mice

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

Abstract

Functional engineering of musculoskeletal tissues generally involves rapid expansion of progenitor cells in vitro while retaining their potential for further differentiation and then induction in specific culture conditions. The autologous adipose-derived stromal cells (ASCs) are considered to contain pluripotent mesenchymal stem cells. Imaging with expression of green fluorescent protein (GFP) facilitates the detailed research on ASCs physiological behavior during differentiation into a variety of cell lineages both in vitro and in vivo. In this study, we aimed to confirm the trans-germ plasticity of homogeneously marked ASCs from GFP transgenic mice. Simultaneously, the term and intensity of GFP expression in ASCs were also focused on during variant inductions, when cells were incubated with multiple growth factors and adjuvant. ASCs were harvested from inguinal fat pads of transgenic nude mice, passaged 3 times in monolayer cultures, and then transferred to osteogenic, adipogenic, neurogenic, and myogenic medium. The morphological characterization of inductive cells was observed using phase-contrast microscopy and histological staining such as alizarin red for mineralization nodules and oil red O for lipid accumulation. The expression of marker genes or proteins was measured using RT-PCR and immunocytochemical analysis. Collagen type I, osteopontin (OPN), and osteocalcin (OCN) were positive in osteogenic lineages, peroxisome proliferator-activated receptor(PPAR)-γ2 and lipoprotein lipase (LPL) were positive in adipogenic ones, glial fibrillary acidic protein (GFAP) and neuron-specific enolase (NSE) were positive in neurogenic ones, and α-smooth muscle actin (α-SMA) was positive in myogenic ones. Moreover, the results of fluorescence microscopic imaging suggested that there was no significant decline of GFP expression during ASCs differentiation and the level of GFP maintained stable till differentiated ASCs showed apoptotic phenotype. So the endogenous GFP and multilineage potential of transgenic ASCs had no influences on each other. Since the population of GFP ASCs can be easily identified, it is proposed that they may be promising candidate seed cells for further studies on ASCs tissue engineering, especially the study on engineered tissues formed in vivo.

Keywords

multilineage differentiation of Adipose-Derived stromal cells from GFP transgenic mice 

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Yunfeng Lin
    • 1
    • 3
  • Xizhe Chen
    • 1
  • Zhengbin Yan
    • 1
  • Lei Liu
    • 1
  • Wei Tang
    • 1
  • Xiaohui Zheng
    • 1
  • Zhiyong Li
    • 1
  • Ju Qiao
    • 2
  • Shengwei Li
    • 1
  • 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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