In Vivo Transfer of Intracellular Labels from Locally Implanted Bone Marrow Stromal Cells to Resident Tissue MacrophagesReport as inadecuate




In Vivo Transfer of Intracellular Labels from Locally Implanted Bone Marrow Stromal Cells to Resident Tissue Macrophages - Download this document for free, or read online. Document in PDF available to download.

Intracellular labels such as dextran coated superparamagnetic iron oxide nanoparticles SPION, bromodeoxyuridine BrdU or green fluorescent protein GFP are frequently used to study the fate of transplanted cells by in vivo magnetic resonance imaging or fluorescent microscopy. Bystander uptake of labeled cells by resident tissue macrophages TM can confound the interpretation of the presence of intracellular labels especially during direct implantation of cells, which can result in more than 70% cell death. In this study we determined the percentages of TM that took up SPION, BrdU or GFP from labeled bone marrow stromal cells BMSCs that were placed into areas of angiogenesis and inflammation in a mouse model known as Matrigel™ plaque perfusion assay. Cells recovered from digested plaques at various time points were analyzed by fluorescence microscopy and flow cytometry. The analysis of harvested plaques revealed 5% of BrdU+, 5–10% of GFP+ and 5–15% of dextran+ macrophages. The transfer of the label was not dependent on cell dose or viability. Collectively, this study suggests that care should be taken to validate donor origin of cells using an independent marker by histology and to assess transplanted cells for TM markers prior to drawing conclusions about the in vivo behavior of transplanted cells.



Author: Edyta Pawelczyk , Elaine K. Jordan, Arun Balakumaran, Aneeka Chaudhry, Nicole Gormley, Melissa Smith, Bobbi K. Lewis, Richard Chi

Source: http://plos.srce.hr/



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