RT Journal Article
SR Electronic
T1 TissueGrinder, a novel technology for rapid generation of patient-derived single cell suspensions from solid tumors by mechanical tissue dissociation
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.06.08.447529
DO 10.1101/2021.06.08.447529
A1 Stefan Scheuermann
A1 Jonas M. Lehmann
A1 Ramkumar Ramani Mohan
A1 Christoph Reißfelder
A1 Felix Rückert
A1 Jens Langejürgen
A1 Prama Pallavi
YR 2021
UL http://biorxiv.org/content/early/2021/06/08/2021.06.08.447529.abstract
AB Introduction 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.Methods 33 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.Results 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.Discussion 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 will be targeted.Competing Interest StatementSS and JL are co-founder of Fast Forward Discoveries GmbH distributing TissueGrinder-Technology. All other authors do not report a conflict of interest.