A10细胞系：一种用于新生儿、新生内膜或分化型血管平滑肌细胞的模型？

The A10 cell line: a model for neonatal, neointimal, or differentiated vascular smooth muscle cells?

作者信息

Rao R S, Miano J M, Olson E N, Seidel C L

机构信息

Baylor College of Medicine, Section of Cardiovascular Sciences, Houston, TX 77030, USA.

出版信息

Cardiovasc Res. 1997 Oct;36(1):118-26. doi: 10.1016/s0008-6363(97)00156-9.

DOI:10.1016/s0008-6363(97)00156-9

PMID:9415280

Abstract

OBJECTIVES

The A10 cell line was derived from the thoracic aorta of embryonic rat and is a commonly used model of vascular smooth muscle cells (VSMC). Despite its wide use this cell line has not been well characterized. This is especially important in light of recent evidence of phenotypically distinct cell populations isolated from rat vascular tissue. Therefore, the present study was undertaken to confirm the VSMC nature of A10 cells and to investigate whether these cells particularly resemble adult, neonatal, or neointimal rat VSMC.

METHODS

A variety of defining characteristics were used that included immunofluorescent analysis for smooth muscle alpha-actin, smooth and non-muscle myosin heavy chains, desmin and vimentin; Western analysis for smooth muscle and non-muscle myosin heavy chains; mRNA analysis for smooth muscle myosin heavy chain, calponin, SM22 alpha, tropoelastin and PDGF-B peptide; and functional assays of cell migration, proliferation and agonist induced intracellular Ca transients.

RESULTS

A10 cells expressed smooth muscle alpha-actin, SM22 alpha, smooth muscle calponin and vimentin, characteristic of in vivo rat VSMCs; however they also resembled de-differentiated smooth muscle cells in that they expressed non-muscle myosin rather than smooth muscle myosin heavy chain. A10 cells resembled cultured rat neonatal smooth muscle cells ("pup cells") in that they had an epithelioid shape and lacked functional PDGF-alpha receptors: however they did not express PDGF-B mRNA or proliferate in low serum containing medium as do neonatal cells. A10 cells had several characteristics in common with neointimal cells including the expression of alpha-actin, vimentin, and non-muscle myosin and the lack of expression of PDGF-B mRNA as well as the ability to migrate in response to PDGF-BB.

CONCLUSION

In conclusion, A10 cells are nondifferentiated VSMC that differ from neonatal but bear significant resemblance to neointimal cells.

摘要

目的

A10细胞系源自胚胎大鼠的胸主动脉，是血管平滑肌细胞（VSMC）常用的模型。尽管该细胞系应用广泛，但其特征尚未得到充分描述。鉴于最近从大鼠血管组织中分离出表型不同的细胞群体的证据，这一点尤为重要。因此，本研究旨在确认A10细胞的VSMC性质，并研究这些细胞是否特别类似于成年、新生或大鼠新生内膜的VSMC。

方法

采用了多种定义特征，包括对平滑肌α-肌动蛋白、平滑肌和非肌肉肌球蛋白重链、结蛋白和波形蛋白进行免疫荧光分析；对平滑肌和非肌肉肌球蛋白重链进行蛋白质免疫印迹分析；对平滑肌肌球蛋白重链、钙调蛋白、SM22α、原弹性蛋白和血小板衍生生长因子B肽进行mRNA分析；以及细胞迁移、增殖和激动剂诱导的细胞内钙瞬变的功能测定。

结果

A10细胞表达平滑肌α-肌动蛋白、SM22α、平滑肌钙调蛋白和波形蛋白，这是体内大鼠VSMC的特征；然而，它们也类似于去分化的平滑肌细胞，因为它们表达非肌肉肌球蛋白而不是平滑肌肌球蛋白重链。A10细胞类似于培养的大鼠新生平滑肌细胞（“幼鼠细胞”），因为它们具有上皮样形状且缺乏功能性血小板衍生生长因子α受体：然而，它们不像新生细胞那样表达血小板衍生生长因子B mRNA或在低血清培养基中增殖。A10细胞与新生内膜细胞有几个共同特征，包括α-肌动蛋白、波形蛋白和非肌肉肌球蛋白的表达，血小板衍生生长因子B mRNA的缺乏表达，以及对血小板衍生生长因子BB的迁移反应能力。