Chick embryonic cells as a source for generating in vitro model of muscle cell dystrophy
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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.
KeywordsChick embryo Embryonic cells Muscle dystrophy In vitro model α-dystroglycan
This work was supported by University Grants Commission, New Delhi, grant number F.No.43–593/2014(SR).
- Grodzki AC, Berenstein E (2010) antibody purification: affinity chromatography–protein a and protein G Sepharose. In: Immunocytochemical methods and protocols. Humana Press, New York, United States, pp 33–41Google Scholar
- Godinho RO (2006) In vitro development of skeletal muscle fiber. Braz J Morphol Sci 23:173–186Google Scholar
- Strober W (2015) Trypan blue exclusion test of cell viability. Curr Protoc Immunol 111(1):A3-BGoogle Scholar