Urja Verma, Khaire Kashmira, Balakrishnan Suresh, Uggini Gowri Kumari
Department of Zoology, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, 390002, India.
In Vitro Cell Dev Biol Anim. 2018 Dec;54(10):756-769. doi: 10.1007/s11626-018-0297-8. Epub 2018 Oct 9.
Chick embryonic cells can be used to develop an easy and economical in vitro model for conducting studies on the disease muscle dystrophy (MD). For this, the limb myoblasts from 11th day chick embryo were isolated and cultured. To this muscle cell culture, anti-dystroglycan antibody (IIH6) was added so as to target the α-dystroglycan and disrupt the connection between the cytoskeletal proteins and the extracellular matrix (which is a characteristic feature of MD). Cells were allowed to differentiate further and the morphometrics and mRNA expression were studied. The IIH6-treated muscle cells displayed changes in morphometry, contractibility, and also atrophy was observed when compared to the control cultures. Concomitant gene expression studies showed an upregulation in TGF-β expression, while the muscle sculpture genes MYOD1, MYF5, LAMA2 and MYOG were downregulated resembling the MD in vivo. This simple and cost-effective method can be useful in studies to further understand the disease mechanism and also in conducting initial studies on effect of novel pharmacological agents.
鸡胚细胞可用于建立一种简单且经济的体外模型,用于开展肌肉萎缩症(MD)的研究。为此,从第11天鸡胚中分离并培养肢体成肌细胞。向这种肌肉细胞培养物中添加抗肌营养不良聚糖抗体(IIH6),以靶向α-肌营养不良聚糖并破坏细胞骨架蛋白与细胞外基质之间的连接(这是MD的一个特征)。使细胞进一步分化,并研究形态计量学和mRNA表达。与对照培养物相比,经IIH6处理的肌肉细胞在形态计量学、收缩性方面表现出变化,并且还观察到萎缩。伴随的基因表达研究表明,TGF-β表达上调,而肌肉形成相关基因MYOD1、MYF5、LAMA2和MYOG下调,类似于体内的MD。这种简单且经济高效的方法可用于进一步了解疾病机制的研究,也可用于开展新型药物制剂效果的初步研究。