The Delivery of Multipotent Adult Progenitor Cells to Extended Criteria Human Donor Livers Using Normothermic Machine Perfusion
The Delivery of Multipotent Adult Progenitor Cells to Extended Criteria Human Donor Livers Using Normothermic Machine Perfusion
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DOI：
10.3389/fimmu.2020.01226
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发表时间：
2020-06-25
影响因子：
7.3
通讯作者：
Afford, Simon C.
中科院分区：
文献类型：
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作者：
Laing, Richard W.;Stubblefield, Samantha;Afford, Simon C.
背景：使用多能成体祖细胞（MAPC（R）细胞，Multistem，Athersys Inc.，克利夫兰，俄亥俄州）表明它们作为器官移植中抗炎和免疫调节疗法的潜力。已经提出了肝脏的常温机器灌注（NMP-L）作为将治疗剂引入供体器官中的方式。使用该技术向人类供体肝脏递送细胞疗法尚未在文献中描述。本研究的主要目的是开发一种使用NMP-L向人类供体肝脏提供细胞治疗的技术，并证明植入。方法：使用肝脏辅助（器官辅助，格罗宁根）在37 ℃下灌注6小时。在灌注期间的不同时间点，将50 X IO 6个CMPTX标记的MAPC细胞经肝动脉（HA，n= 3）或门静脉（PV，n= 3）在20分钟内直接输注到右叶中。记录灌注参数，并在多个时间点从两个肺叶进行中央和外周活检，并进行标准组织学染色和共聚焦显微镜检查。使用35重多重测定和蛋白质组学分析来分析灌注液。结果：通过任一途径输注MAPC细胞对灌注流参数没有不利影响。六个肝脏中有三个符合器官活力的既定标准。共聚焦显微镜显示，通过动脉灌注时，MAPC细胞穿过血管内皮植入。35-对灌流液进行多重分析，得到13个阳性靶点，其中9个与MAPC细胞输注有关（包括白细胞介素1b、4、5、6、8、10、MCP-1、GM-CSF、SDF-1a）。蛋白质组学分析揭示了细胞治疗输注后时间点的灌注液中的295种独特蛋白质，其中许多与文献中的MAPC细胞和间充质干细胞有很强的联系。功能富集分析证明了它们的免疫调节潜力。结论：我们已经证明，细胞可以直接输送到靶器官，在宿主免疫细胞群暴露之前，并且不会影响灌注。动脉输注后发生跨内皮迁移。MAPC细胞似乎分泌一系列可溶性因子，这些因子在人类肝移植模型中具有抗炎和免疫调节益处。
Background:Pre-clinical research with multi-potent adult progenitor cells (MAPC (R) cells, Multistem, Athersys Inc., Cleveland, Ohio) suggests their potential as an anti-inflammatory and immunomodulatory therapy in organ transplantation. Normothermic machine perfusion of the liver (NMP-L) has been proposed as a way of introducing therapeutic agents into the donor organ. Delivery of cellular therapy to human donor livers using this technique has not yet been described in the literature. The primary objectives of this study were to develop a technique for delivering cellular therapy to human donor livers using NMP-L and demonstrate engraftment. Methods:Six discarded human livers were perfused for 6 h at 37 degrees C using the Liver Assist (Organ Assist, Groningen). 50 x 106 CMPTX-labeled MAPC cells were infused directly into the right lobe via the hepatic artery (HA,n= 3) or portal vein (PV,n= 3) over 20 min at different time points during the perfusion. Perfusion parameters were recorded and central and peripheral biopsies were taken at multiple time-points from both lobes and subjected to standard histological stains and confocal microscopy. Perfusate was analyzed using a 35-plex multiplex assay and proteomic analysis. Results:There was no detrimental effect on perfusion flow parameters on infusion of MAPC cells by either route. Three out of six livers met established criteria for organ viability. Confocal microscopy demonstrated engraftment of MAPC cells across vascular endothelium when perfused via the artery. 35-plex multiplex analysis of perfusate yielded 13 positive targets, 9 of which appeared to be related to the infusion of MAPC cells (including Interleukin's 1b, 4, 5, 6, 8, 10, MCP-1, GM-CSF, SDF-1a). Proteomic analysis revealed 295 unique proteins in the perfusate from time-points following the infusion of cellular therapy, many of which have strong links to MAPC cells and mesenchymal stem cells in the literature. Functional enrichment analysis demonstrated their immunomodulatory potential. Conclusion:We have demonstrated that cells can be delivered directly to the target organ, prior to host immune cell population exposure and without compromising the perfusion. Transendothelial migration occurs following arterial infusion. MAPC cells appear to secrete a host of soluble factors that would have anti-inflammatory and immunomodulatory benefits in a human model of liver transplantation.