五指山猪二尖瓣间质细胞的分离、培养与鉴定

Isolation, culture and identification of mitral valveinterstitial cells from Wuzhishan pig

黏液瘤样二尖瓣疾病(MMVD)是哺乳动物最常见的心脏病之一,临床上可引发心力衰竭和猝死,目前该病的发病机制尚不完全明确。在MMVD发生发展进程中,二尖瓣间质细胞(MVICs)活化引起的细胞外基质重塑是本病的关键病理变化。因此本研究拟通过建立五指山猪原代MVICs分离、培养体系,获得可稳定培养的MVICs,为MMVD相关研究提供体外研究细胞材料。结果表明,用目前方法分离得到的五指山猪二尖瓣细胞呈CD31阴性、Vimentin阳性,符合MVICs特点。对比添加不同浓度血清的培养基,含10%胎牛血清(FBS)的完全培养基对MVICs细胞增殖活力更有利,可支持MVICs在体外稳定培养至少20代。本研究获得的五指山猪原代MVICs可为二尖瓣疾病病理机制研究与进一步的靶向药物开发提供体外研究平台。

Myxomatous mitral valve disease(MMVD)is one of the most common heart diseases in mammals,which can lead to heart failure and sudden death in clinical practice.The extracellular matrix remodeling caused by mitral stromal cells(MVICs)activation is the key pathological change in the development of MMVD.Therefore,this study intended to establish a separation and culture system for primary MVICs of Wuzhishan pig to obtain stable cultured MVICs,providing cell materials for in vitro research on MMVD.The results showed that the mitral valve cells of Wuzhishan pig isolated by the present method were CD31 negative and Vimentin positive,which were consistent with the characteristics of MVIC.Compared with the medium with different serum concentrations,the complete medium containing 10%FBS serum was more favorable to the proliferation activity of MVICs cells,and could support the stable culture of MVICs in vitro for at least 20 generations.The primary MVICs of Wuzhishan pig obtained in this study can provide an in vitro research platform for the study of the pathological mechanism of mitral valve disease and the further development of targeted drugs.