Scheuermann Stefan, Lehmann Jonas M, Ramani Mohan Ramkumar, Reißfelder Christoph, Rückert Felix, Langejürgen Jens, Pallavi Prama
Clinical Health Technologies, Fraunhofer IPA, Mannheim, Germany.
Department of Surgery, Universitätsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
Front Med (Lausanne). 2022 Dec 1;9:721639. doi: 10.3389/fmed.2022.721639. eCollection 2022.
Recent advances hold promise of making personalized medicine a step closer to implementation in clinical settings. However, traditional sample preparation methods are not robust and reproducible. In this study, the TissueGrinder, a novel mechanical semi-automated benchtop device, which can isolate cells from tissue in a very fast and enzyme-free way is tested for cell isolation from surgically resected tumor tissues.
Thirty-three surgically resected tumor tissues from various but mainly pancreatic, liver or colorectal origins were processed by both novel TissueGrinder and explant method. An optimized processing program for tumors from pancreatic, liver or colorectal cancer was developed. The viability and morphological characteristics of the isolated cells were evaluated microscopically. Expression of pancreatic cancer markers was evaluated in cells isolated from pancreatic tumors. Finally, the effect of mechanical stress on the cells was evaluated by assessing apoptosis markers via western blotting.
TissueGinder was more efficient in isolating cells from tumor tissue with a success rate of 75% when compared to explant method 45% in terms of cell outgrowth six weeks after processing. Cells isolated with TissueGinder had a higher abundance and were more heterogeneous in composition as compared to explant method. Mechanical processing of the cells with TissueGrinder does not lead to apoptosis but causes slight stress to the cells.
Our results show that TissueGrinder can process solid tumor tissues more rapidly and efficiently and with higher success rate compared to the conventionally used explant method. The results of the study suggest that the TissueGrinder might be a suitable method for obtaining cells, which is important for its application in individualized therapy. Due to the great variance in different tumor entities and the associated individual tissue characteristics, a further development of the dissociation protocol for other types of tumors and normal tissue will be targeted.
近期的进展有望使个性化医疗在临床环境中的实施更近一步。然而，传统的样本制备方法并不稳定且缺乏可重复性。在本研究中，对新型机械半自动台式设备TissueGrinder进行了测试，该设备能够以非常快速且无酶的方式从组织中分离细胞，用于从手术切除的肿瘤组织中分离细胞。
采用新型TissueGrinder和外植体方法对33个手术切除的肿瘤组织进行处理，这些肿瘤组织来源各异，但主要来自胰腺、肝脏或结肠直肠。针对胰腺癌、肝癌或结直肠癌肿瘤开发了优化的处理程序。通过显微镜评估分离细胞的活力和形态特征。对从胰腺肿瘤中分离的细胞评估胰腺癌标志物的表达。最后，通过蛋白质印迹法评估凋亡标志物，以评价机械应力对细胞的影响。
与外植体方法相比，TissueGrinder在从肿瘤组织中分离细胞方面更有效，处理后六周细胞生长成功率为75%，而外植体方法为45%。与外植体方法相比，用TissueGrinder分离的细胞丰度更高，组成更具异质性。用TissueGrinder对细胞进行机械处理不会导致细胞凋亡，但会对细胞造成轻微应激。
我们的结果表明，与传统使用的外植体方法相比，TissueGrinder能够更快速、高效地处理实体肿瘤组织，成功率更高。研究结果表明，TissueGrinder可能是一种获取细胞的合适方法，这对其在个体化治疗中的应用很重要。由于不同肿瘤实体存在很大差异以及相关的个体组织特征，将针对其他类型肿瘤和正常组织的解离方案进行进一步开发。
