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两种高产分离大鼠主动脉平滑肌细胞的方法。

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

作者信息

Lotfollahzadeh Saran, Jose Asha, Zarnaab Shafiq Esha, El Sherif Nourhan, Smith Michael, Han Jingyan, Seta Francesca, Chitalia Vipul

机构信息

Renal Section, Department of Medicine, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, United States.

Department of Biomedical Engineering, College of Engineering, Boston University, Boston, MA 02118, United States.

出版信息

Biol Methods Protoc. 2024 May 22;9(1):bpae038. doi: 10.1093/biomethods/bpae038. eCollection 2024.

Abstract

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.

摘要

血管平滑肌细胞(VSMCs)是血管的重要组成部分,也是血管生物学、转化研究和心血管疾病深入研究的重点。虽然有永生化的血管平滑肌细胞系,但它们的应用有限,这凸显了对原代VSMCs的需求。有几种从小鼠中分离原代细胞的方法。然而,鉴于小鼠和大鼠主动脉的差异,从大鼠血管中分离的方法需要优化。在这里,我们比较了两种从大鼠中分离VSMCs的方法:酶消化法和“块”法。我们观察到使用酶消化法时VSMCs的产量显著更高。我们进一步证实,两种方法分离的VSMCs均表达成熟的VSMC特异性标志物(钙调蛋白),并观察到该标志物在传至9代时仍持续存在,这表明VSMCs的分泌表型得以延续。总体而言,这项工作比较了两种方法,并展示了一种从大鼠主动脉中分离VSMCs的实用有效方法,为血管生物学家提供了一个有价值且可靠的实验工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/85dd/11244693/109be7b1a6eb/bpae038f1.jpg

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