TY - JOUR
T1 - The A10 cell line
T2 - A model for neonatal, neointimal, or differentiated vascular smooth muscle cells?
AU - Rao, Rohini S.
AU - Miano, Joseph M.
AU - Olson, Eric N.
AU - Seidel, Charles L.
N1 - Funding Information:
We thank Dr. Julius C. Allen and Dr. Mark W. Majesky for helpful discussion and for critically reading the manuscript. Dr. William P. Schilling is gratefully acknowledged for his advice and for the use of equipment in the intracellular calcium measurements. Work by RSR and CLS was supported in part by a grant from the American Heart Association (910-13100). RSR is a student in the Graduate Program in Cardiovascular Sciences, Baylor College of Medicine. ENO is funded by the National Institutes of Health, Muscular Dystrophy Association and the Robert A. Welch Foundation. JMM is a recipient of an NRSA Postdoctoral Fellowship.
Funding Information:
Analysis of variance (ANOVA) with the student-Newman-Keuls post test was used for Fig. 7 and Fig. 8. Computational assistance was provided by the CLINFO project, supported by the Division of Research Resources of the NIH under grant number RR-00350.
PY - 1997/10
Y1 - 1997/10
N2 - 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 α-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, SM22α, tropoelastin and PDGF-B peptide: and functional assays of cell migration, proliferation and agonist induced intracellular Ca transients. Results: A10 cells expressed smooth muscle α- actin, SM22α, 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-α 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 α-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.
AB - 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 α-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, SM22α, tropoelastin and PDGF-B peptide: and functional assays of cell migration, proliferation and agonist induced intracellular Ca transients. Results: A10 cells expressed smooth muscle α- actin, SM22α, 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-α 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 α-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.
KW - A10 cells
KW - Cell migration
KW - Cell proliferation
KW - Cytoskeletal proteins
KW - PDGF
KW - Rat
KW - Vascular smooth muscle
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U2 - 10.1016/S0008-6363(97)00156-9
DO - 10.1016/S0008-6363(97)00156-9
M3 - Article
C2 - 9415280
AN - SCOPUS:0030656973
SN - 0008-6363
VL - 36
SP - 118
EP - 126
JO - Cardiovascular Research
JF - Cardiovascular Research
IS - 1
ER -