Isolation of Human Primary Valve Cells for In vitro Disease Modeling.
Isolation of Human Primary Valve Cells for In vitro Disease Modeling.
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DOI：
10.3791/62439
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发表时间：
2021-04-16
期刊：
影响因子：
--
通讯作者：
St Hilaire C
中科院分区：
文献类型：
--
作者：
Cuevas RA;Chu CC;Moorhead WJ 3rd;Wong R;Sultan I;St Hilaire C
钙化性主动脉瓣疾病（CAVD）存在于近三分之一的老年人中。主动脉瓣增厚、硬化和钙化导致主动脉狭窄，并导致心力衰竭和中风。疾病的发病机制是多因素的，炎症、细胞外基质重塑、湍流、机械应力和应变等应激有助于瓣膜内皮细胞和瓣膜间质细胞的成骨分化。然而，驱动健康细胞向钙化细胞的成骨转变的确切启动因素尚未完全确定。此外，目前唯一治疗cavd诱导的主动脉瓣狭窄的方法是主动脉瓣置换术，即移除原有瓣膜（外科主动脉瓣置换术，SAVR）或通过导管插入完全可折叠的置换瓣膜（经导管主动脉瓣置换术，TAVR）。这些外科手术费用高，风险大；因此，为药物发现确定新的治疗靶点是必要的。为此，本研究开发了一种工作流程，将手术切除的患者组织和供体尸体组织用于创建用于体外疾病建模的患者特异性瓣膜细胞原代系。该方案引入了器官移植中常用的冷藏溶液的使用，以减少组织切除和实验室处理之间通常较长的采购时间所造成的损害，并大大稳定了切除组织的细胞。本研究的结果表明，离体瓣膜细胞在从供体取下瓣膜后数天内仍能保持其增殖能力和内皮和间质表型。使用这些材料可以收集控制细胞和CAVD细胞，从这些细胞中建立控制和疾病细胞系。
Calcific aortic valve disease (CAVD) is present in nearly a third of the elderly population. Thickening, stiffening, and calcification of the aortic valve causes aortic stenosis and contributes to heart failure and stroke. Disease pathogenesis is multifactorial, and stresses such as inflammation, extracellular matrix remodeling, turbulent flow, and mechanical stress and strain contribute to the osteogenic differentiation of valve endothelial and valve interstitial cells. However, the precise initiating factors that drive the osteogenic transition of a healthy cell into a calcifying cell are not fully defined. Further, the only current therapy for CAVD-induced aortic stenosis is aortic valve replacement, whereby the native valve is removed (surgical aortic valve replacement, SAVR) or a fully collapsible replacement valve is inserted via a catheter (transcatheter aortic valve replacement, TAVR). These surgical procedures come at a high cost and with serious risks; thus, identifying novel therapeutic targets for drug discovery is imperative. To that end, the present study develops a workflow where surgically removed tissues from patients and donor cadaver tissues are used to create patient-specific primary lines of valvular cells for in vitro disease modeling. This protocol introduces the utilization of a cold storage solution, commonly utilized in organ transplant, to reduce the damage caused by the often-lengthy procurement time between tissue excision and laboratory processing with the benefit of greatly stabilizing cells of the excised tissue. The results of the present study demonstrate that isolated valve cells retain their proliferative capacity and endothelial and interstitial phenotypes in culture upwards of several days after valve removal from the donor. Using these materials allows for the collection of control and CAVD cells, from which both control and disease cell lines are established.
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影响因子：
2
作者：
Sider KL;Blaser MC;Simmons CA
通讯作者：
Simmons CA
影响因子：
5
作者：
Anstine, Lindsey J.;Bobba, Chris;Lincoln, Joy
通讯作者：
Lincoln, Joy
影响因子：
5.6
作者：
Hilaire, Cynthia St.;Carroll, Shannon H.;Chen, Hongjie;Ravid, Katya
通讯作者：
Ravid, Katya
影响因子：
4.6
作者：
Jian, B;Narula, N;Levy, RJ
通讯作者：
Levy, RJ
DOI：
10.1016/s1357-2725(02)00100-0
发表时间：
2003-02-01
影响因子：
4
作者：
Taylor, PA;Batten, P;Yacoub, MH
通讯作者：
Yacoub, MH