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Human Aortic Valve Interstitial Cells Obtained From Patients With Calcified Aortic Valve Stenosis Are Vegfr2 Positive And Contribute To Ectopic Calcification
Zaiqiang Yu, Kazuyuki Daitoku, Masahito Minakawa, Kazuhiko Seya, Tadaatsu Imaizumi.
Hirosaki University, Aomori, Japan.

OBJECTIVE: Calcified aortic valve stenosis (AVS) was the most common heart valve disease in aging society, but there was no medical therapy for inhibiting the acceleration of AVS. Although we recently demonstrated that human aortic valve interstitial cells (HAVICs) obtained from patients with AVS were highly sensitive to ectopic calcification stimulation, the cell types contributing to aortic valve ectopic calcification are unknown. We aimed to immunocytochemically characterize HAVICs and identify their contribution to valve ectopic calcification.
METHODS: This study was approved by the institutional review boards of the Hospital of Hirosaki University. Calcified aortic valves were obtained from AVS patients performed surgical aortic valve replacement (SAVR), who were given sufficient informed consent preoperative. Human aortic valve interstitial cells (HAVICs) were isolated from aortic valves of patients with AVS by collagenase and cultured on non-coated plastic dishes.
RESULTS: Immunocytochemical features and HAVIC differentiation activity were analyzed in passage 1 (P1). Most cultured P1 HAVICs were CD73-, CD90-, and CD105-positive, but CD45- and CD34-negative. We further confirmed that cultured P1 HAVICs showed vascular endothelial growth factor receptor 2 (VEGFR2)-positive, which did not disappeared according to cell proliferation; However, approximately half of cultured P1 HAVICs were α-smooth muscle actin (SMA)-positive, colonized, and easily differentiated into osteoblastic cells, which was confirmed. Furtherly, VEGFR2-positive HAVICs obtained from patients with calcified AVS were more sensitive to tumor necrosis factor-α-induced valve ectopic calcification, but which had no relationship with or without α-SMA positivity.
CONCLUSIONS: These data suggest that HAVICs obtained from patients with calcified AVS are VEGFR2-positive undifferentiated mesenchymal cells and may contribute to aortic valve ectopic calcification. It is very important for medical innovation of AVS by targeting VEGFR2 in future.


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