Abstract
Non-invasive monitoring of living cells in vivo provides an important tool in the development of cell-based therapies in cartilage tissue engineering. High-resolution magnetic resonance imaging (MRI) has been used to monitor target cell populations in vivo. However, the side-effects on cell function of the labelling reagents, such as superparamagnetic iron oxide (SPIO), are still unclear. This study investigated the effect of SPIO particles on the chondrogenic differentiation of human bone marrow stromal cells (HBMSCs), neonatal and adult chondrocytes in vitro. Cells were labelled with SPIO for 24 h and chondrogenesis induced in serum-free medium including TGFβ3. For labelled/unlabelled cells, viability, morphology and proliferation were determined using CellTracker™ Green and PicoGreen dsDNA assays. The expression of SOX9, COL2A1 and ACAN was investigated using qRT–PCR after 2, 7 and 14 days. The results showed that viability was unaffected in all of the cells but cell morphology changed towards a 'stretched' phenotype following SPIO uptake. Cell proliferation was reduced only for labelled neonatal chondrocytes. SOX9 and COL2A1 expression decreased at day 2 but not at days 7 and 14 for labelled HBMSCs and adult chondrocytes; ACAN expression was unaffected. In contrast, SOX9 and COL2A1 expression were unaffected in labelled neonatal chondrocytes but a decrease in ACAN expression was seen at day 14. The results suggest that downregulation of chondrogenic genes associated with SPIO labelling is temporary and target cell-dependent. Resovist® can be used to label HBMSCs or mature chondrocytes for MR imaging of cells for cartilage tissue engineering. Copyright © 2012 John Wiley & Sons, Ltd.
Highlights
Cell therapy is an emerging tool in regenerative medicine
Our previous publication showed that glyceraldehyde3-phosphate dehydrogenase (GAPDH) was a reliable housekeeping gene, as its expression levels did not differ between experimental and control samples for the three cell types (Saha et al, 2010)
The effects of Resovist without the use of a transfection agents (TAs) have been investigated on the chondrogenic potential of cells from different sources that might be used for cartilage repair (e.g. human bone marrow stromal cells (HBMSCs), human neonatal chondrocytes and human adult chondrocytes) (Saha et al, 2010)
Summary
Cell therapy is an emerging tool in regenerative medicine. Bioluminescence, radioactive labelling and near-infrared fluorescence are some of the techniques that have been used to determine cell fate after applications of cell-based therapies by tracking homing and migration of the transplanted cells (Thorne et al, 2006; Contag, 2006; Hofmann et al, 2005; Scarff et al, 2006). A variety of SPIOs are available for use as MRI contrast agents. Thereafter, a number of studies showed that transfection agents (TAs), such as protamine, poly-L-lysine and lipofectamine, interact electrostatically with the SPIO. These are an effective and stable means to non-covalently bind to DNA and internalize the MR label into the cells (Frank et al, 2003; Kostura et al, 2004). A number of SPIO particles that do not require the assistance of TAs, such as ResovistW (generic name, ferucarbotran; Schering, Germany) and FeridexW, have been made commercially available (Mailander et al, 2008). Mailander et al, (2008) showed Resovist to label cells more efficiently than Feridex without a TA (Mailander et al, 2008). Resovist is approved for clinical use in Europe, Japan and Australia (Vogl et al, 1996; Lutz et al, 2005; Wersebe et al, 2006)
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