Traditionally, muscle contraction is triggered by electric field stimulation using electrodes placed in extracellular space. Alternatively, spontaneously beating cardiomyocytes have been used. Electric field stimulation is a simple method to control the time pattern of systolic activity. However, the electric field is usually uneven and stimulates many unexpected muscle cells at the same time. Therefore, it is difficult to specifically stimulate the identified muscle cell population.
Compared with traditional electrical stimulation methods, light stimulation methods have attracted much attention because of their fine spatial and temporal resolution, parallel stimulation of multiple parts, relatively harmless and convenient.
Construction of myoblast cell line which stably express ChR2 can be achieved at Creative BioMart. It allows people to stimulate muscle cells without contact with tissue or extracellular fluid. Because the optical signal also has high spatial and temporal resolution, it should enable people to accurately control muscle contraction in a given spatial and temporal mode.
Cell Culture and Generation of ChR2 viral-Transfected C2C12 Cell Lines
A murine leukemia virus based vector was constructed to mediate the expression of ChR2. The carrier has a modified yellow fluorescent protein. In order to produce a stable cell line expressing ChR2, myoblasts were transfected with virus particles and cloned twice by limited dilution. Single cell clones with bright Venus fluorescence were selected under conventional inverted fluorescence microscope.
Formation and Maturation of Myotubes
Myoblasts were induced to differentiate into myotubes in the differentiation medium composed of DMEM.
This technology will have many potential bioengineering applications, such as wireless driving of muscle dynamic actuators/micro devices, nerve culture model system that can be used to study muscle fiber maturation, muscle bioassay of biological and mechanical properties, and pathological and physiological research of muscular dystrophy, This method will be able to produce human muscle tissue substitutes whose contraction is optically regulated.
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