Triple-modal imaging of stem-cells labeled with multimodal nanoparticles, applied in a stroke model

• 作者: Andrea Tiemi Kondo Daianne Maciely Carvalho Fantacini Dimas Tadeu Covas Fernando Anselmo de Oliveira Helio Rodrigues da Silva Javier Bustamante Mamani José Mauro Kutner Leopoldo Penteado Nucci Lionel Fernel Gamarra Lucas Eduardo Botelho de Souza Mariana Penteado Nucci Nelson Hamerschlak Virginia Picanao-Castro
• 来源期刊: 世界干细胞杂志:英文版(电子版)
• 学科: 医学
• 关键词: MULTIMODAL NANOPARTICLES Human bone MARROW mesenchymal stem cells Near-infrared fluorescence image Magnetic resonance BIOLUMINESCENCE STROKE
• 年，卷（期）: 2019,（2）
• 页码范围: 100-123
• 页数: 24页
• 分类号: R

摘要

BACKGROUND Mesenchymal stem cells (MSCs) have been widely tested for their therapeutic efficacy in the ischemic brain and have been shown to provide several benefits. A major obstacle to the clinical translation of these therapies has been the inability to noninvasively monitor the best route, cell doses, and collateral effects while ensuring the survival and effective biological functioning of the transplanted stem cells. Technological advances in multimodal imaging have allowed in vivo monitoring of the biodistribution and viability of transplanted stem cells due to a combination of imaging technologies associated with multimodal nanoparticles (MNPs) using new labels and covers to achieve low toxicity and longtime residence in cells. AIM To evaluate the sensitivity of triple-modal imaging of stem cells labeled with MNPs and applied in a stroke model. METHODS After the isolation and immunophenotypic characterization of human bone marrow MSCs (hBM-MSCs), our team carried out lentiviral transduction of these cells for the evaluation of bioluminescent images (BLIs) in vitro and in vivo. In addition, MNPs that were previously characterized (regarding hydrodynamic size, zeta potential, and optical properties), and were used to label these cells, analyze cell viability via the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay and BLI analysis, and quantify the internalization process and iron load in different concentrations of MNPs via magnetic resonance imaging (MRI), near-infrared fluorescence (NIRF), and inductively coupled plasma-mass spectrometry (ICP-MS). In in vivo analyses, the same labeled cells were implanted in a sham group and a stroke group at different times and under different MNP concentrations (after 4 h or 6 d of cell implantation) to evaluate the sensitivity of triple-modal images. RESULTS hBM-MSC collection and isolation after immunophenotypic characterization were demonstrated to be adequate in hBM samples. After transduction of these cells with luciferase (hBM-MSCLuc), we detec