Study Of The Influence Of Optical And Electrical Stimulation On The Growth And Proliferation Of Myoblasts

Myoblast is the precursor cell of cytoplasm containing myofilament. It is asingle-layer and adherent growth cell and can spontaneously split and proliferate.Myoblast has the ability of self-renewal and damage reparation. Compared with othercells, it has obvious advantages in gene therapy. Thus, myoblast plays an important rolein clinical and basic research. Its growth is closely related to temperature, osmoticpressure, PH, electricity and light. Among them, light and electricity can significantlycontrol the speed of the proliferation and differentiation of cells, so the light andelectrical stimulation is always the research hot spot in in-vitro cell culture. So far, thereare a large number of experimental studies related to the light and electrical stimulationin clinical and basic research. Some of them have achieved certain results and aregradually moving towards the industrialization and clinical application. For example,light therapy is being used for leukoderma, psoriasis and other skin diseases.Electrotherapy is used for sciatica and many kinds of painful diseases.On the background, this paper will study the effect of light and electric stimulationon the proliferation of myoblasts through setting up home-made auxiliary devices. Themajor work is:①Read the literature and analyze the theoretical basis for the experimentalfeasibility. Before carrying out the experiment, many related experimental studies havebeen gone through such as the light stimulation experiment of cells by using low-energylasers. LED was selected as light source to study the biological effect of lightstimulation on fibroblast. Another one is the electrical stimulation of neural stem cell bymicro electric current pulse. It discussed the effect of electrical stimulation on theproliferation and differentiation of neural stem cells through loading micro-electriccurrent pulse on self-made electrodes. The mechanism of light and electrical stimulationcould be achieved by discussing these experimental results, and relevant theoreticalbasis can be put forward by analyzing the feasibility of this paper.②Design and building of the light and electrical stimulation device. Based on thetheoretical basis of the previous discussion, light and electric devices are respectivelydesigned. Low cost and power consumption are also thought of. Three kinds of LEDs (88) with a wavelength of467nm,589nm and630nm are selected as light sources tobuild a LED array through the combination of series and parallel style. Stainless steel is chosen as the electrode material, and PDMS is selected as the pouring solidificationmaterial to build the electric stimulation device. Plate electrodes (7cm1cm) areselected, and the distance between two electrodes is4.5cm.③Cell culture and stimulation experiments. After experimental plan is decided,stimulation parameters such as timing, number, wavelength and time are determined. Inthe culture of myoblasts, when cells have covered the plate (commonly75%~80%), asubculture will be achieved and the number of subcultures will be determined accordingto the experimental requirement. Stimulated timing are12h or24h after inoculation,stimulated numbers are one, two, three or four, stimulated wavelengths are467nm,589nm or630nm, and stimulated time is400s,600s,1000s or1600s. Above parametersare respectively used in turn to discuss their effect on the proliferation of myoblasts.④Evaluation of the growth status of myoblasts and determination of optimalstimulation parameters. After stimulation experiments, cells are collected and fixed forthe detection of cell cycle after24hours. In collecting and fixing processes, digestionusing pancreatic enzyme should not be excessive. Otherwise, cells will be broken toproduce debris. Centrifugal speed should be set to1000rpm. If the speed is too slow,cells cannot be very well coagulated to lead the loss of cells. On the contrary, if thespeed is too fast, cells will be broken. Cellular debris and broken cells will affect theresults of the flow cytometric detection. Furthermore, the number of cells cannot be lessthan1～2×10~6. Comparing with the experimental results with those of Guo Ruixiong, aset of optimal stimulation parameters can be determined.