Efficient transfection of mouse-derived C2C12 myoblasts using a matrigel basement membrane matrix.
Myogenic cell lines have been used widely in the study of myogenic differentiation, muscle regeneration and homeostasis, but, myoblasts and myotubes are difficult to transfect using conventional techniques. We have used liposome-based transfection method to introduce a green fluorescence protein (GFP)-expressing plasmid into Matrigel basement membrane matrix-coated C2C12 mouse myoblast cells. Myoblasts adhered and proliferated more rapidly on a Matrigel; thus, a dramatic increase in transfection efficiency can be obtained compared to Matrigel-untreated cells. Transfection efficiency was determined by counting fluorescent and total cells from six random fields for each condition. This protocol results in efficient (up to 60-70%) transfection of C2C12 myoblasts, high levels of GFP expression and low rate of cell death (10%). This technique is rapid, reliable, uses a lipid-based transfection reagent, and yields high transfection rates in a previously hard-to-transfect cell type.
Balci
Burcu
B
Hacettepe University, Faculty of Medicine, Department of Medical Biology, Sihhiye, Ankara, Turkey.
Dinçer
Pervin
P
D000818:Animals; D001485:Basement Membrane / Q000737:chemistry / Q000378:metabolism; D048708:Cell Growth Processes / Q000502:physiology; D002460:Cell Line; D003094:Collagen / Q000737:chemistry / Q000378:metabolism*; D004338:Drug Combinations; D007797:Laminin / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D032446:Myoblasts / Q000378:metabolism / Q000502:physiology*; D010784:Photomicrography; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen