Timraz Sara B H, Farhat Ilyas A H, Alhussein Ghada, Christoforou Nicolas, Teo Jeremy C M
Department of Biomedical Engineering, Khalifa University, PO Box 127788, Abu Dhabi, United Arab Emirates.
Department of Applied Mathematics and Sciences, Khalifa University, PO Box 127788, Abu Dhabi, United Arab Emirates.
Exp Cell Res. 2016 May 1;343(2):168-176. doi: 10.1016/j.yexcr.2016.04.004. Epub 2016 Apr 11.
In vitro research on vascular tissue engineering has extensively used isolated primary human or animal smooth muscle cells (SMC). Research programs that lack such facilities tend towards commercially available primary cells sources. Here, we aim to evaluate the capacity of commercially available human SMC to maintain their contractile phenotype, and determine if dedifferentiation towards the synthetic phenotype occurs in response to conventional cell culture and passaging without any external biochemical or mechanical stimuli. Lower passage SMC adopted a contractile phenotype marked by a relatively slower proliferation rate, higher expression of proteins of the contractile apparatus and smoothelin, elongated morphology, and reduced deposition of collagen types I and III. As the passage number increased, migratory capacity was enhanced, average cell speed, total distance and net distance travelled increased up to passage 8. Through the various assays, corroborative evidence pinpoints SMC at passage 7 as the transition point between the contractile and synthetic phenotypes, while passage 8 distinctly and consistently exhibited characteristics of synthetic phenotype. This knowledge is particularly useful in selecting SMC of appropriate passage number for the target vascular tissue engineering application, for example, a homeostatic vascular graft for blood vessel replacement versus recreating atherosclerotic blood vessel model in vitro.
血管组织工程的体外研究广泛使用分离的原代人或动物平滑肌细胞(SMC)。缺乏此类设施的研究项目倾向于使用市售的原代细胞来源。在此,我们旨在评估市售人SMC维持其收缩表型的能力,并确定在没有任何外部生化或机械刺激的情况下,对传统细胞培养和传代培养是否会发生向合成表型的去分化。传代次数较低的SMC呈现收缩表型,其特征为增殖速率相对较慢、收缩装置和平滑肌蛋白的表达较高、形态细长以及I型和III型胶原蛋白沉积减少。随着传代次数增加,迁移能力增强,直至第8代,平均细胞速度、总移动距离和净移动距离均增加。通过各种检测,确凿证据表明第7代SMC是收缩表型和合成表型之间的转变点,而第8代明显且一致地表现出合成表型的特征。这一知识对于为目标血管组织工程应用选择合适传代次数的SMC尤为有用, 例如,用于血管置换的稳态血管移植物与在体外重建动脉粥样硬化血管模型。