Imaging cellular pharmacokinetics of 18F-FDG and 6-NBDG uptake by inflammatory and stem cells.
Imaging cellular pharmacokinetics of 18F-FDG and 6-NBDG uptake by inflammatory and stem cells.
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DOI：
10.1371/journal.pone.0192662
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Xing L
中科院分区：
文献类型：
--
作者：
Zaman RT;Tuerkcan S;Mahmoudi M;Saito T;Yang PC;Chin FT;McConnell MV;Xing L
心肌梗死（MI）导致心肌细胞的显著损失、心肌组织损伤和心肌功能损害。心肌细胞不能增殖，阻止了心脏的自我再生。心肌梗死后晚期心力衰竭的治疗是心脏移植，尽管器官的可用性有限。因此，基于干细胞的心脏疗法可以通过修复受损的心肌来逆转心肌细胞的损失，从而最终预防心力衰竭。然而，基于干细胞的疗法缺乏对成功疗法背后机制的理解，包括难以追踪干细胞以提供有关细胞迁移、增殖和分化的信息。在这项研究中，我们研究了不同类型的干细胞和炎症细胞与细胞靶向成像分子18 F-FDG和6-NBDG之间的相互作用，以确定体外摄取模式和药代动力学。将巨噬细胞（M1和M2）、人诱导多能干细胞（hiPSC）和人羊膜间充质干细胞（hAMSC）与18 F-FDG或6-NBDG孵育。去除过量的放射性示踪剂和荧光，将100 μ m薄的CdWO4闪烁体板置于细胞顶部，用于18F-FDG摄取的放射发光显微镜成像，而6-NBDG摄取的荧光成像不需要闪烁体。β衰变后产生的光用高灵敏度倒置显微镜（LV 200，Olympus）和电子倍增电荷耦合器件（EM-CCD）照相机成像。在MATLAB中开发了用于图像处理的定制软件。与hiPSC（0.540 ± 0.026 fCi/μ m2，P = 0.003）和巨噬细胞（0.430 ± 0.023 fCi/μ m2，P = 0.002）中的摄取相比，hAMSC的单个细胞中18F-FDG的平均细胞活性（0.670 ± 0.028 fCi/μ m2，P = 0.001）分别高20%和36%。hAMSC表现出最慢的流入（0.210 min-1），但最快的流出（0.327 min-1）的18 F-FDG相比，其他测试的细胞系。该细胞系也具有最高的磷酸化，但表现出最低的去磷酸化速率。在这三种细胞系中，6-NBDG的摄取模式非常不同。巨噬细胞单个细胞中6-NBDG的平均细胞活性（0.570 ± 0.230 fM/μ m2，P = 0.004）分别比hiPSC（0.350 ± 0.160 fM/μ m2，P = 0.001）和hAMSC（0.490 ± 0.028 fM/μ m2，P = 0.006）高38%和14%。与其他两种测试的细胞系相比，巨噬细胞的流入（0.276 min-1）、流出（0.612 min-1）、磷酸化（0.269 min-1）和去磷酸化（0.049 min-1）速率也最高。发现hAMSC对18F-FDG分子的敏感性是hiPSC/巨噬细胞的2 - 3倍。然而，巨噬细胞对6-NBDG表现出最大的敏感性。基于这一结果，18F-FDG靶向的hAMSCs可能更适合于通过收集细胞迁移、增殖和分化的信息来了解成功治疗MI患者的机制。
Myocardial infarction (MI) causes significant loss of cardiomyocytes, myocardial tissue damage, and impairment of myocardial function. The inability of cardiomyocytes to proliferate prevents the heart from self-regeneration. The treatment for advanced heart failure following an MI is heart transplantation despite the limited availability of the organs. Thus, stem-cell-based cardiac therapies could ultimately prevent heart failure by repairing injured myocardium that reverses cardiomyocyte loss. However, stem-cell-based therapies lack understanding of the mechanisms behind a successful therapy, including difficulty tracking stem cells to provide information on cell migration, proliferation and differentiation. In this study, we have investigated the interaction between different types of stem and inflammatory cells and cell-targeted imaging molecules, 18F-FDG and 6-NBDG, to identify uptake patterns and pharmacokinetics in vitro. Macrophages (both M1 and M2), human induced pluripotent stem cells (hiPSCs), and human amniotic mesenchymal stem cells (hAMSCs) were incubated with either 18F-FDG or 6-NBDG. Excess radiotracer and fluorescence were removed and a 100 μm-thin CdWO4 scintillator plate was placed on top of the cells for radioluminescence microscopy imaging of 18F-FDG uptake, while no scintillator was needed for fluorescence imaging of 6-NBDG uptake. Light produced following beta decay was imaged with a highly sensitive inverted microscope (LV200, Olympus) and an Electron Multiplying Charge-Couple Device (EM-CCD) camera. Custom-written software was developed in MATLAB for image processing. The average cellular activity of 18F-FDG in a single cell of hAMSCs (0.670±0.028 fCi/μm2, P = 0.001) was 20% and 36% higher compared to uptake in hiPSCs (0.540±0.026 fCi/μm2, P = 0.003) and macrophages (0.430±0.023 fCi/μm2, P = 0.002), respectively. hAMSCs exhibited the slowest influx (0.210 min-1) but the fastest efflux (0.327 min-1) rate compared to the other tested cell lines for 18F-FDG. This cell line also has the highest phosphorylation but exhibited the lowest rate of de-phosphorylation. The uptake pattern for 6-NBDG was very different in these three cell lines. The average cellular activity of 6-NBDG in a single cell of macrophages (0.570±0.230 fM/μm2, P = 0.004) was 38% and 14% higher compared to hiPSCs (0.350±0.160 fM/μm2, P = 0.001) and hAMSCs (0.490±0.028 fM/μm2, P = 0.006), respectively. The influx (0.276 min-1), efflux (0.612 min-1), phosphorylation (0.269 min-1), and de-phosphorylation (0.049 min-1) rates were also highest for macrophages compared to the other two tested cell lines. hAMSCs were found to be 2–3× more sensitive to 18F-FDG molecule compared to hiPSCs/macrophages. However, macrophages exhibited the most sensitivity towards 6-NBDG. Based on this result, hAMSCs targeted with 18F-FDG could be more suitable for understanding the mechanisms behind successful therapy for treating MI patients by gathering information on cell migration, proliferation and differentiation.
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影响因子：
3.3
作者：
Arai, T;Kofidis, T;Yang, PC
通讯作者：
Yang, PC
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影响因子：
20.1
作者：
Tachibana A;Santoso MR;Mahmoudi M;Shukla P;Wang L;Bennett M;Goldstone AB;Wang M;Fukushi M;Ebert AD;Woo YJ;Rulifson E;Yang PC
通讯作者：
Yang PC
影响因子：
17.1
作者：
Hong H;Yang K;Zhang Y;Engle JW;Feng L;Yang Y;Nayak TR;Goel S;Bean J;Theuer CP;Barnhart TE;Liu Z;Cai W
通讯作者：
Cai W
影响因子：
3.7
作者：
Pratx G;Chen K;Sun C;Martin L;Carpenter CM;Olcott PD;Xing L
通讯作者：
Xing L
影响因子：
20.1
作者：
Kim PJ;Mahmoudi M;Ge X;Matsuura Y;Toma I;Metzler S;Kooreman NG;Ramunas J;Holbrook C;McConnell MV;Blau H;Harnish P;Rulifson E;Yang PC
通讯作者：
Yang PC