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

IF 2.5 Q3 BIOCHEMICAL RESEARCH METHODS Biology Methods and Protocols Pub Date : 2024-05-22 DOI:10.1093/biomethods/bpae038
Saran Lotfollahzadeh, Asha Jose, Esha Shafiq zarnaab, Nourhan El Sherif, Michael Smith, Jingyan Han, Francesca Seta, V. Chitalia
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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.
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两种高产率分离大鼠主动脉平滑肌细胞的方法
血管平滑肌细胞(VSMC)是血管不可或缺的组成部分,也是血管生物学、转化研究和心血管疾病领域深入研究的重点。虽然有永生化的血管平滑肌细胞系,但其用途有限,因此需要原代血管平滑肌细胞。从小鼠体内分离原代细胞有多种方法。然而,由于小鼠和大鼠的主动脉不同,从大鼠血管中分离的方法需要优化。在此,我们比较了从大鼠体内分离 VSMC 的两种方法:酶解法和 "块状 "法。我们观察到酶解法的 VSMC 得率明显更高。我们还进一步证实,这两种方法都能使 VSMC 表达成熟的 VSMC 特异性标记(钙蛋白),并观察到这一标记可持续到 9 次传代,这表明 VSMC 的分泌表型仍在继续。总之,这项工作对两种方法进行了比较,展示了一种从大鼠主动脉分离 VSMC 的实用有效方法,为血管生物学家提供了一种宝贵可靠的实验工具。
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来源期刊
Biology Methods and Protocols
Biology Methods and Protocols Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)
CiteScore
3.80
自引率
2.80%
发文量
28
审稿时长
19 weeks
期刊最新文献
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