Two methods of isolation of rat aortic smooth muscle cells with high yield.

Autor:	Lotfollahzadeh S; Renal Section, Department of Medicine, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States., Jose A; Renal Section, Department of Medicine, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States., Zarnaab Shafiq E; Renal Section, Department of Medicine, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States., El Sherif N; Department of Biomedical Engineering, College of Engineering, Boston University, Boston, MA 02118, United States., Smith M; Department of Biomedical Engineering, College of Engineering, Boston University, Boston, MA 02118, United States., Han J; Whitaker Cardiovascular Institute, Department of Medicine, Vascular Biology Section, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States., Seta F; Whitaker Cardiovascular Institute, Department of Medicine, Vascular Biology Section, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States., Chitalia V; Renal Section, Department of Medicine, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States.; Veterans Affairs Boston Healthcare System, Boston, MA 02118, United States.; Center of Cross-Organ Vascular Pathology, Department of Medicine, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States.
Jazyk:	angličtina
Zdroj:	Biology methods & protocols [Biol Methods Protoc] 2024 May 22; Vol. 9 (1), pp. bpae038. Date of Electronic Publication: 2024 May 22 (Print Publication: 2024).
DOI:	10.1093/biomethods/bpae038
Abstrakt:	Vascular smooth muscle cells (VSMCs) are an integral part of blood vessels and are the focus of intensive research in vascular biology, translational research, and cardiovascular diseases. Though immortalized vascular smooth muscle cell lines are available, their use is limited, underscoring the need for primary VSMCs. There are several methods for isolating primary cells from mice. However, the isolation method from rat blood vessels requires optimization, given the differences in the aorta of mice and rats. Here we compare two methods for VSMCs isolation from rats: enzymatic digestion and the "block" method. We observed a significantly higher yield of VSMCs using the enzymatic digestion method. We further confirmed that VSMCs expressed well-established VSMC-specific markers (calponin) with both methods and observed the persistence of this marker up to 9 passages, suggesting a continuation of the secretory phenotype of VSMCs. Overall, this work compares two methods and demonstrates a practical and effective method for isolating VSMCs from rat aorta, providing vascular biologists with a valuable and reliable experimental tool. (© The Author(s) 2024. Published by Oxford University Press.)
Databáze:	MEDLINE
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