In vitro model of bromodeoxyuridine or iron oxide nanoparticle uptake by activated macrophages from labeled stem cells: Implications for cellular therapy
In vitro model of bromodeoxyuridine or iron oxide nanoparticle uptake by activated macrophages from labeled stem cells: Implications for cellular therapy
复制标题
DOI：
10.1634/stemcells.2007-0707
复制
发表时间：
2008-05-01
期刊：
影响因子：
5.2
通讯作者：
Frank, Joseph A.
中科院分区：
文献类型：
--
作者：
Pawelczyk, Edyta;Arbab, Ali S.;Frank, Joseph A.
有越来越多的兴趣使用外源性标记，如溴脱氧尿苷（BrdU）或超顺磁性氧化铁纳米粒子（SPION）标记细胞，以识别移植的细胞和监测其迁移的荧光显微镜或在体内磁共振成像（MRI），分别。将细胞直接植入靶组织可导致>80%的细胞由于创伤或凋亡而死亡。激活的巨噬细胞（AM）对标记细胞的旁观者摄取可能混淆结果的解释。本研究使用Boyden室炎症模型研究了AM吸收BrdU或SPION的频率。将SPION/BrdU标记的骨髓基质细胞或HeLa细胞、AM和小鼠成纤维细胞（MF）或人成纤维细胞（HF）以各种比例在上室的Matrigel中混合，并孵育长达96小时。AM被化学诱导迁移到下室。在MF或HF存在下，使用抗-CD 68、抗-BrdU、抗-葡聚糖抗体对来自下室和上室的AM进行荧光激活细胞分选分析，在孵育96小时后显示10%-20%葡聚糖阳性或10% BrdU阳性AM。铁转移到AM占
There is increasing interest in using exogenous labels such as bromodeoxyuridine (BrdU) or superparamagnetic iron oxide nanoparticles (SPION) to label cells to identify transplanted cells and monitor their migration by fluorescent microscopy or in vivo magnetic resonance imaging (MRI), respectively. Direct implantation of cells into target tissue can result in >80% cell death due to trauma or apoptosis. Bystander uptake of labeled cells by activated macrophages (AM) can confound the interpretation of results. This study investigated the frequency of BrdU or SPION uptake by AM using the Boyden chamber model of inflammation. SPION/BrdU-Iabeled bone marrow stromal cells or HeLa cells, AM, and mouse fibroblasts (MF) or human fibroblasts (HF) were mixed in various ratios in Matrigel in the upper chamber and incubated for up to 96 hours. The AM were chemotactically induced to migrate to the lower chamber. Fluorescence-activated cell sorting analysis of AM from lower and upper chambers, in the presence of either MF or HF using anti-CD68, anti-BrdU, anti-dextran antibodies, revealed 10%-20% dextran-positive or 10% BrdU-positive AM after 96 hours of incubation. Transfer of iron to AM accounted for