Pham Phuc H, Vo Nguyen T K, Tan Elizabeth J H, Russell Spencer, Jones Ginny, Lumsden John S, Bols Niels C
Department of Biology, University of Waterloo, Waterloo, ON, Canada.
Department of Medical Physics and Applied Radiation Sciences, McMaster University, Hamilton, ON, Canada.
In Vitro Cell Dev Biol Anim. 2017 Jan;53(1):20-32. doi: 10.1007/s11626-016-0077-2. Epub 2016 Sep 1.
As diseases and abnormalities of the heart can interfere with the aquaculture of Atlantic salmon, the heart was investigated as a source of cell lines that could be used to study the cellular basis of these conditions. An Atlantic salmon heart endothelial cell line, ASHe, was developed and characterized for growth properties, endothelial cell characteristics, and responsiveness to lysophosphatidic acid (LPA). AHSe cells stained negative for senescence associated ß-galactosidase and grew well in 10 and 20% FBS/L15 at high cell density, but not in L15 medium supplemented with calf serum. It displayed many endothelial cell-like characteristics including a cobblestone morphology, capillary-like structures formation on Matrigel, and expression of von Willebrand factor and endothelial cell-related tight junction proteins ZO-1, claudin 3, and claudin 5. ASHe cells responded to the cardiovascular modulator, LPA, in two contrasting ways. LPA at 5 and 25 μM inhibited the ability of ASHe cells to heal a wound but stimulated their proliferation, especially as evaluated by colony formation in low-density cultures. The enhancement of proliferation by LPA parallels what has been observed previously in mammalian endothelial cell cultures exposed to LPA, whereas the LPA slowing of ASHe cell migration contrasted with the LPA-enhanced migration of some mammalian cells. Therefore, this cell line is a potentially useful model for future comparative studies on piscine and mammalian cardiovascular cell biology and for studies on diseases of Atlantic salmon in aquaculture.
由于心脏疾病和异常会干扰大西洋鲑的养殖，因此对心脏进行了研究，以寻找可用于研究这些病症细胞基础的细胞系来源。开发了一种大西洋鲑心脏内皮细胞系ASHe，并对其生长特性、内皮细胞特征以及对溶血磷脂酸（LPA）的反应性进行了表征。AHSe细胞衰老相关β-半乳糖苷酶染色呈阴性，在高细胞密度下于10%和20%胎牛血清/L15培养基中生长良好，但在添加小牛血清的L15培养基中生长不佳。它表现出许多内皮细胞样特征，包括鹅卵石样形态、在基质胶上形成毛细血管样结构，以及表达血管性血友病因子和内皮细胞相关紧密连接蛋白ZO-1、claudin 3和claudin 5。ASHe细胞对心血管调节剂LPA有两种截然不同的反应。5 μM和25 μM的LPA抑制ASHe细胞愈合伤口的能力，但刺激其增殖，尤其是通过低密度培养中的集落形成来评估时。LPA对增殖的增强与先前在暴露于LPA的哺乳动物内皮细胞培养物中观察到的情况相似，而LPA减缓ASHe细胞迁移则与某些哺乳动物细胞中LPA增强的迁移形成对比。因此，该细胞系对于未来关于鱼类和哺乳动物心血管细胞生物学的比较研究以及大西洋鲑养殖疾病的研究是一个潜在有用的模型。
