Rose Laura C, Kadayakkara Deepak K, Wang Guan, Bar-Shir Amnon, Helfer Brooke M, O'Hanlon Charles F, Kraitchman Dara L, Rodriguez Ricardo L, Bulte Jeff W M
Division of Magnetic Resonance Research, Russell H. Morgan Department of Radiology and Radiological Science, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA Cellular Imaging Section and Vascular Biology Program, Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Division of Magnetic Resonance Research, Russell H. Morgan Department of Radiology and Radiological Science, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA Cellular Imaging Section and Vascular Biology Program, Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Stem Cells Transl Med. 2015 Dec;4(12):1472-81. doi: 10.5966/sctm.2015-0113. Epub 2015 Oct 28.
Stromal vascular fraction (SVF) cells are used clinically for various therapeutic targets. The location and persistence of engrafted SVF cells are important parameters for determining treatment failure versus success. We used the GID SVF-1 platform and a clinical protocol to harvest and label SVF cells with the fluorinated ((19)F) agent CS-1000 as part of a first-in-human phase I trial (clinicaltrials.gov identifier NCT02035085) to track SVF cells with magnetic resonance imaging during treatment of radiation-induced fibrosis in breast cancer patients. Flow cytometry revealed that SVF cells consisted of 25.0% ± 15.8% CD45+, 24.6% ± 12.5% CD34+, and 7.5% ± 3.3% CD31+ cells, with 2.1 ± 0.7 × 10⁵ cells per cubic centimeter of adipose tissue obtained. Fluorescent CS-1000 (CS-ATM DM Green) labeled 87.0% ± 13.5% of CD34+ progenitor cells compared with 47.8% ± 18.5% of hematopoietic CD45+ cells, with an average of 2.8 ± 2.0 × 10¹² ¹⁹F atoms per cell, determined using nuclear magnetic resonance spectroscopy. The vast majority (92.7% ± 5.0%) of CD31+ cells were also labeled, although most coexpressed CD34. Only 16% ± 22.3% of CD45-/CD31-/CD34- (triple-negative) cells were labeled with CS-ATM DM Green. After induction of cell death by either apoptosis or necrosis, >95% of ¹⁹F was released from the cells, indicating that fluorine retention can be used as a surrogate marker for cell survival. Labeled-SVF cells engrafted in a silicone breast phantom could be visualized with a clinical 3-Tesla magnetic resonance imaging scanner at a sensitivity of approximately 2 × 10⁶ cells at a depth of 5 mm. The current protocol can be used to image transplanted SVF cells at clinically relevant cell concentrations in patients.
Stromal vascular fraction (SVF) cells harvested from adipose tissue offer great promise in regenerative medicine, but methods to track such cell therapies are needed to ensure correct administration and monitor survival. A clinical protocol was developed to harvest and label SVF cells with the fluorinated (¹⁹F) agent CS-1000, allowing cells to be tracked with (19)F magnetic resonance imaging (MRI). Flow cytometry evaluation revealed heterogeneous ¹⁹F uptake in SVF cells, confirming the need for careful characterization. The proposed protocol resulted in sufficient ¹⁹F uptake to allow imaging using a clinical MRI scanner with point-of-care processing.
基质血管成分（SVF）细胞在临床上用于各种治疗靶点。植入的SVF细胞的位置和持久性是决定治疗失败或成功的重要参数。我们使用GID SVF-1平台和一项临床方案，用氟化（¹⁹F）试剂CS-1000收获并标记SVF细胞，这是一项首次在人体进行的I期试验（clinicaltrials.gov标识符NCT02035085）的一部分，目的是在乳腺癌患者放射性纤维化治疗期间用磁共振成像追踪SVF细胞。流式细胞术显示，SVF细胞由25.0%±15.8%的CD45⁺细胞、24.6%±12.5%的CD34⁺细胞和7.5%±3.3%的CD31⁺细胞组成，每立方厘米脂肪组织获得2.1±0.7×10⁵个细胞。荧光CS-1000（CS-ATM DM Green）标记了87.0%±13.5%的CD34⁺祖细胞，而造血CD45⁺细胞的标记率为47.8%±18.5%，使用核磁共振波谱法测定，每个细胞平均有2.8±2.0×10¹²个¹⁹F原子。绝大多数（92.7%±5.0%）的CD31⁺细胞也被标记，尽管大多数同时表达CD34。只有16%±22.3%的CD45⁻/CD31⁻/CD34⁻（三阴性）细胞被CS-ATM DM Green标记。在通过凋亡或坏死诱导细胞死亡后，>95%的¹⁹F从细胞中释放出来，这表明氟保留可以用作细胞存活的替代标志物。植入硅胶乳房模型中的标记SVF细胞可以用临床3特斯拉磁共振成像扫描仪在5毫米深度处大约2×10⁶个细胞的灵敏度下进行可视化。当前方案可用于在患者中以临床相关细胞浓度对移植的SVF细胞进行成像。
从脂肪组织中收获的基质血管成分（SVF）细胞在再生医学中具有巨大潜力，但需要追踪此类细胞疗法的方法来确保正确给药并监测存活情况。开发了一项临床方案，用氟化（¹⁹F）试剂CS-1000收获并标记SVF细胞，从而能够用¹⁹F磁共振成像（MRI）追踪细胞。流式细胞术评估显示SVF细胞中¹⁹F摄取存在异质性，证实了仔细表征的必要性。所提出的方案导致了足够的¹⁹F摄取，以便使用具有即时护理处理功能的临床MRI扫描仪进行成像。
