Influence of ultrasonic stimulation on the growth and proliferation of Oryza sativa Nipponbare callus cells

Abstract

Ultrasound acts as an alternative stress on cells or tissues. In this study it is aimed to investigate the effect of ultrasonic stimulation on the growth and proliferation of Oryza sativa Nipponbare cells (rice callus) in suspension culture. After the samples were stimulated by ultrasound at 28 kHz, we measured their growth and proliferation by using a colorimetric MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay as well as fresh weights of the cells. Growth curves were obtained by fresh weights in the suspension culture after ultrasonic agitation for different time from 2 to 120 s. In MTT method, the optical density was determined at 570 nm in the cell suspension on 10 days after the ultrasonic agitation. Up to 5 s agitation OD 570 increased; it decreased for more prolonged stimulation. We found that ultrasonic stimulation could promote the growth and proliferation of O. sativa Nipponbare cells in suspension culture with the optimal stimulation of 5 s, while with longer agitation, its growth and proliferation was inhibited. The mechanism may be that the ultrasound activated or destroyed the cellular structure, such as cell membrane, cytoskeleton and mitochondria in which many enzymes and ion channels are affected. In addition, the enhancement of cell wall and cell membrane fluidity might be one of the factors to promote the cell growth in 5-s ultrasonic stimulation.