Ultrasound frequency sonication facilitates high-throughput and uniform dissociation of cellular aggregates and tissues.

Autor:	Welch EC; School of Engineering, Center for Biomedical Engineering, Brown University, United States., Chaltas K; School of Engineering, Center for Biomedical Engineering, Brown University, United States., Tripathi A; School of Engineering, Center for Biomedical Engineering, Brown University, United States. Electronic address: Anubhav_Tripathi@brown.edu.
Jazyk:	angličtina
Zdroj:	SLAS technology [SLAS Technol] 2023 Apr; Vol. 28 (2), pp. 70-81. Date of Electronic Publication: 2023 Jan 12.
DOI:	10.1016/j.slast.2023.01.001
Abstrakt:	A sample preparation step involving dissociation of tissues into their component cells is often required to conduct analysis of nucleic acids and other constituents from tissue samples. Frequently, the extracellular matrix and cell-cell adhesions are disrupted via treatment with a chemical dissociating reagent or various mechanical forces. In this work, a new, high-throughput, multiplexed method of dissociating tissues and cellular aggregates into single cells using ultrasound frequency bath sonication is explored and characterized. Different operating parameters are evaluated, and a treatment protocol with potential for uniform, high-throughput tissue dissociation is compared to the existing best chemical and orbital plate shaking protocol. Metrics such as percent dissociation, cellular recovery, average aggregate size, proportion of various aggregate sizes, membrane circularity, and cellular viability are subsequently assessed and found to be favorable. In optimized conditions, 53 ± 8% of 1 mm biopsy cores are dissociated within 30 min using sonication alone, surpassing leading high-throughput orbital plate shaking techniques five-fold. Chemical digestion is also 2 times more effective when complexed with sonication rather than orbital plate shaking. RNA content, quality, and expression are found to be superior to the standard protocol in terms of transcriptional preservation. Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Anubhav Tripathi reports financial support was provided by PerkinElmer Inc. Anubhav Tripathi reports a relationship with PerkinElmer Inc that includes: consulting or advisory and funding grants. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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