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Cardiovascular Research 1997 36(1):118-126; doi:10.1016/S0008-6363(97)00156-9
© 1997 by European Society of Cardiology
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Copyright © 1997, European Society of Cardiology

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

Rohini S Raoa, Joseph M Mianob, Eric N Olsonc and Charles L Seidela,*

aBaylor College of Medicine, Section of Cardiovascular Sciences, Houston, TX 77030, USA
bMedical College of Wisconsin, Department of Physiology, Houston, TX, USA
cUT Southwestern Medical Center, the Hamon Cntr. for Basic Cancer Research, Houston, TX, USA

* Corresponding author. Dept. of Medicine, Section of Cardiovascular Sciences, Room 512C, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA. Tel.: +1-713-798-4977; fax: +1-713- 790-0681.

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.

KEYWORDS A10 cells; Rat; Vascular smooth muscle; Cytoskeletal proteins; PDGF; Cell migration; Cell proliferation


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