Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (10): 1759-1763.doi: 10.3969/j.issn.1673-8225.2011.10.011

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Culture and molecular imaging identification of adipose derived mesenchymal stem cells with stable reporter gene expression in vitro and in vivo

Fan Wei-wei, Wang Ya-bin, Zhang Rong-qing, Li Cong-ye, Li Shuang, Cao Feng   

  1. Department of Cardiology, Xijing Hospital, Fourth Military Medical University of Chinese PLA, Xi’an  710032, Shaanxi Province, China
  • Received:2010-10-15 Revised:2010-11-16 Online:2011-03-05 Published:2011-03-05
  • Contact: Cao Feng, Chief physician, Professor, Doctoral supervisor, Department of Cardiology, Xijing Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China wind8828@gmail.com
  • About author:Fan Wei-wei★, Studying for master’s degree, Physician, Department of Cardiology, Xijing Hospital, Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China fww2142008458@gmail.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30970845*; the Key Assist Project of Xijing Hospital, No. XJZT08Z04*

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Abstract:

BACKGROUND: Previous studies show that molecular imaging can monitor the stem cell qualitatively and quantitatively at cellular and molecular levels in vivo and hold promise in long-time evaluation of the stem cell.
OBJECTIVE: To construct adipose derived mesenchymal stem cells (ADMSCs) with stable reporter genes expression and identify it by reporter gene imaging in vitro and in vivo.
METHODS: β-actin-luc transgenic mice were selected firstly.Then ADMSCs were cultured from β-actin-luc mice by modified collagenase method. ADMSCs at passage 3 were identified by flow cytometry. ADMSCs (1×106) were implanted into the muscles of hindlimb of BALB/c nude mice, then tracked and quantified by firefly luciferase (fluc) bioluminescence imaging (BLI) in vitro and in vivo.
RESULTS AND CONCLUSION: The transgenic mice stably carried β-actin-luc reporter gene and the ADMSCs were positive for CD90 and CD44, while negative for CD45, CD34 and CD31. ADMSCs consistently expressed the fluc and a robust correlation existed between different cell numbers and fluc average radiance (r2=0.96). After 24-hours engrafted ADMSCs survived and displayed strong BLI signal, which increased rapidly and then decreased smoothly during 0-42 minutes after peritoneal injection of D-luciferin, the peak being (6.92×106±4.11×105) Photons?s-1?cm-2?sr-1 at 21 minutes. Results indicated that ADMSCs of β-actin-luc transgenic mice could highly express the markers of MSCs, stably carry the reporter gene and facilitate the tracking and quantifying by BLI in vitro and in vivo.

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