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细胞迁移的正电子发射断层扫描成像:一种细胞放射性标记方法

英文原题:Positron Emission Tomography Imaging of Cell Trafficking: A Method of Cell Radiolabeling.

查看英文原题

Positron Emission Tomography Imaging of Cell Trafficking: A Method of Cell Radiolabeling.

PubMed 2023/10/27(内容时间) J Vis Exp Q3 · IF 1.2(JCR 2025)

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中文摘要

干细胞和嵌合抗原受体(CAR)T细胞疗法正成为器官再生及多种癌症免疫治疗的有前景方法。尽管相关领域已取得显著进展,仍需进一步研究,以更好地了解治疗细胞在活体内的药代动力学和药效学。为使用正电子发射断层成像(PET)对细胞进行无创体内追踪,研究人员开发了一种由89Zr介导的细胞放射性标记方法,使用[89Zr]Zr-p-异硫氰酸苄基去铁胺([89Zr]Zr-DBN;89Zr半衰期78.4小时)。本方案描述一种即用型[89Zr]Zr-DBN放射性标记合成子,可直接标记多种细胞,包括间充质干细胞、谱系定向心脏生成干细胞、肝再生肝细胞、白细胞、黑色素瘤细胞及树突状细胞。所建立的方法可在给药后最多7天内通过PET无创成像追踪细胞迁移,且不影响放射性标记细胞的性质或功能。

此外,本方案还分步介绍[89Zr]Zr-DBN的放射合成、其生物相容性制剂的配制、细胞放射标记前的准备,以及使用[89Zr]Zr-DBN标记细胞的过程,并提供成功完成细胞标记所需的详细操作信息。

展开英文摘要原文

Stem cell and chimeric antigen receptor (CAR) T-cell therapies are emerging as promising therapeutics for organ regeneration and as immunotherapy for various cancers. Despite significant progress having been made in these areas, there is still more to be learned to better understand the pharmacokinetics and pharmacodynamics of the administered therapeutic cells in the living system. For noninvasive, in vivo tracking of cells with positron emission tomography (PET), a novel [ 89 Zr]Zr-p-isothiocyanatobenzyl-desferrioxamine ([ 89 Zr]Zr-DBN)-mediated cell radiolabeling method has been developed utilizing 89 Zr (t1/2 78.

4 h). The present protocol describes a [ 89 Zr]Zr-DBN-mediated, ready-to-use, radiolabeling synthon for direct radiolabeling of variety of cells, including mesenchymal stem cells, lineage-guided cardiopoietic stem cells, liver regenerating hepatocytes, white blood cells, melanoma cells, and dendritic cells. The developed methodology enables noninvasive PET imaging of cell trafficking for up to 7 days post-administration without affecting the nature or the function of the radiolabeled cells.

Additionally, this protocol describes a stepwise method for the radiosynthesis of [ 89 Zr]Zr-DBN, biocompatible formulation of [ 89 Zr]Zr-DBN, preparation of cells for radiolabeling, and finally the radiolabeling of cells with [ 89 Zr]Zr-DBN, including all the intricate details needed for the successful radiolabeling of cells.

论文信息

作者
Bansal A、DeGrado TR、Pandey MK
第一作者单位
Division of Nuclear Medicine, Department of Radiology, Mayo Clinic.
通讯作者单位
Division of Nuclear Medicine, Department of Radiology, Mayo Clinic; Pandey.Mukesh@mayo.edu.
文献类型
音视频资料 · 非美国政府资助研究 · 美国 NIH 资助研究
期刊
Journal of visualized experiments : JoVE2023 Oct 27
原文标识
PubMed 37955388 · DOI 10.3791/64117