Investigating Cancerous Exosomes' Effects on CD8+ T-Cell IL-2 Production in a 3D Unidirectional Flow Bioreactor Using 3D Printed, RGD-Functionalized PLLA Scaffolds.
Investigating Cancerous Exosomes' Effects on CD8+ T-Cell IL-2 Production in a 3D Unidirectional Flow Bioreactor Using 3D Printed, RGD-Functionalized PLLA Scaffolds.
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使用3D打印的RGD官能化的PLLA支架研究了3D单向流生物反应器中癌性外泌体对CD8+ T细胞IL-2产生的影响。
DOI：
10.3390/jfb13010030
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发表时间：
2022-03-11
影响因子：
4.8
通讯作者：
Sikavitsas VI
中科院分区：
文献类型：
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作者：
Karami D;Srivastava A;Ramesh R;Sikavitsas VI
来自癌细胞的外泌体与癌症的进展和转移有关，其携带的免疫抑制因子限制了免疫细胞的抗肿瘤能力。实时、3D细胞/支架结构流灌注系统的开发已被探索作为研究癌细胞t细胞和外泌体的新工具。将人肺癌细胞（H1299和A549）外泌体与CD8+ t细胞固定在3d打印的rgd功能化聚乳酸（PLLA）支架上，在单向流动生物反应器中共培养，并评估IL-2的产生。IL-2的产生研究了广泛的t细胞与外泌体的比例。通过将RGD结合基元以可控的密度成功地结合到PLLA表面，CD8+ t细胞成功地附着在2D磁盘和3D打印多孔PLLA支架上。t细胞附着随着RGD表面密度的增加而增加。当RGD密度低于0.5 nmol /mm2时，附着t细胞直径为7.2±0.2µm，当RGD密度为2 nmol /mm2时，由于过度拥挤，附着t细胞直径降至5.1±0.3µm。癌细胞外泌体数量越多，表面附着的t细胞产生的IL-2就越少。在二维磁盘中，静态条件下，当t细胞与外泌体的比例高于1:10时，IL-2的产生被抑制。静态3D多孔支架中IL-2的产生沉默要求比例高于1:20。血流的掺入导致中度至显著的t细胞脱离。以0.15、1.5 mL/min的灌注流量暴露4 h后，3D支架上保留的t细胞比例分别为89±11%和30±8%。在3D支架上，当流速为0.15 ml/min时，当t细胞与外泌体的比例为1:1000时，H1299和A549癌性外泌体均显著抑制IL-2的产生。与静态条件相比，在单向流动条件下培养的t细胞需要更高水平的外泌体来抑制IL-2的产生，这表明培养条件和水动力环境对CD8+ t细胞与癌症外泌体之间的相互作用的重要性。
Exosomes from cancer cells are implicated in cancer progression and metastasis, carrying immunosuppressive factors that limit the antitumor abilities of immune cells. The development of a real-time, 3D cell/scaffold construct flow perfusion system has been explored as a novel tool in the study of T-cells and exosomes from cancer cells. Exosomes from human lung cancer (H1299 and A549) cells were co-cultured in a unidirectional flow bioreactor with CD8+ T-cells immobilized onto 3D-printed RGD-functionalized poly(L-lactic) acid (PLLA) scaffolds and assessed for IL-2 production. The IL-2 production was investigated for a wide range of T-cell to exosome ratios. With the successful incorporation of the RGD binding motif onto the PLLA surface at controllable densities, CD8+ T-cells were successfully attached onto 2D disks and 3D printed porous PLLA scaffolds. T-cell attachment increased with increasing RGD surface density. The diameter of the attached T-cells was 7.2 ± 0.2 µm for RGD densities below 0.5 nmoles/mm2 but dropped to 5.1 ± 0.3 µm when the RGD density was 2 nmoles/mm2 due to overcrowding. The higher the number of cancer exosomes, the less the IL-2 production by the surface-attached T-cells. In 2D disks, the IL-2 production was silenced for T-cell to exosome ratios higher than 1:10 in static conditions. IL-2 production silencing in static 3D porous scaffolds required ratios higher than 1:20. The incorporation of flow resulted in moderate to significant T-cell detachment. The portions of T-cells retained on the 3D scaffolds after exposure for 4 h to 0.15 or 1.5 mL/min of perfusion flow were 89 ± 11% and 30 ± 8%, respectively. On 3D scaffolds and in the presence of flow at 0.15 ml/min, both H1299 and A549 cancerous exosomes significantly suppressed IL-2 production for T-cell to exosome ratios of 1:1000. The much higher level of exosomes needed to silence the IL-2 production from T-cells cultured under unidirectional flow, compared to static conditions, denotes the importance of the culturing conditions and the hydrodynamic environment, on the interactions between CD8+ T-cells and cancer exosomes.
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影响因子：
46.9
作者：
Alvarez-Erviti, Lydia;Seow, Yiqi;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.
影响因子：
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影响因子：
3.7
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Chen WX;Liu XM;Lv MM;Chen L;Zhao JH;Zhong SL;Ji MH;Hu Q;Luo Z;Wu JZ;Tang JH
通讯作者：
Tang JH
影响因子：
16.6
作者：
Domcke, Silvia;Sinha, Rileen;Levine, Douglas A.;Sander, Chris;Schultz, Nikolaus
通讯作者：
Schultz, Nikolaus
影响因子：
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作者：
Anand, Paras K
通讯作者：
Anand, Paras K