Gene transfection in primary stem-like cells of giant cell tumor of bone

Abstract

The neoplastic stem-like stromal cell of giant cell tumor of bone (GCT) survives for multiple passages in primary culture with a stable phenotype, and exhibits multipotent characteristics. The pathophysiology of this tumor has been studied through the primary culture of these cells. However, successful gene transfer of these cells has not been reported to date. In this short report, we describe the development of the first reported technique that results in efficient gene transfection in primary stem-like cells of GCT.

Keywords: TWIST; gene; giant cell tumor; primary cells; transfection.

Figure 1

Green fluorescence protein (GFP) monitored in transfected giant cell tumor of bone stromal and KC cells. Representative immunofluorescence microscopy of KC and GCT-transfected cells with eGFP vector. DAPI staining (blue) indicates the nuclei. (Oil immersion 100× magnification).

Figure 2

TWIST expression monitored in transfected giant cell tumor of bone stromal and KC cells. Representative immunofluorescence microscopy of KC and giant cell tumor of bone transfected cells with TWIST. DAPI staining (blue) indicates the nuclei. TWIST expression was detected using anti-V5 rabbit polyclonal antibody (1:250 dilutions, oil immersion 100× magnification).

Figure 3

TWIST mRNA expression. A) Real-time polymerase chain reaction of cDNA from giant cell tumor of bone stromal cells representing TWIST upregulation in transfected cells relative to TWIST untransfected cells. B) Quantitative real-time polymerase chain reaction expression of TWIST in TWIST transfected (KCT) and untransfected (KC) KC cells. Results are the average of three replicate experiments.