Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (25): 3949-3955.doi: 10.12307/2021.003

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Low-intensity pulsed ultrasound promotes the proliferation and adhesion of human adipose-derived mesenchymal stem cells

Chen Yang, Huang Denggao, Gao Yuanhui, Wang Shunlan, Cao Hui, Zheng Linlin, He Haowei, Luo Siqin, Xiao Jingchuan, Zhang Yingai, Zhang Shufang   

  1. Affiliated Haikou Hospital of Xiangya Medicine College, Central South University, Haikou 570208, Hainan Province, China
  • Received:2020-06-20 Revised:2020-06-28 Accepted:2020-07-23 Online:2021-09-08 Published:2021-03-24
  • Contact: Huang Denggao, Associate researcher, Affiliated Haikou Hospital of Xiangya Medicine College, Central South University, Haikou 570208, Hainan Province, China Zhang Shufang, Researcher, Professor, Affiliated Haikou Hospital of Xiangya Medicine College, Central South University, Haikou 570208, Hainan Province, China
  • About author:Chen Yang, Associate chief technician, Affiliated Haikou Hospital of Xiangya Medicine College, Central South University, Haikou 570208, Hainan Province, China
  • Supported by:
    the Natural Science Foundation of Hainan Province, No. 819QN386 (to HDG), No. 819QN390 (to GYH), No. 18A200137 (to ZYA); Major Scientific and Technological Project in Hainan Province, No. ZDZX2013003 (to ZSF), No. ZDKJ2017007 (to ZSF)

Abstract: BACKGROUND: Mesenchymal stem cells are one of the hot topics in tissue engineering and regenerative medicine. However, how to effectively expand the stem cells in vitro to meet the clinical needs is still a challenge for researchers.
OBJECTIVE: To investigate the effect of low-intensity pulsed ultrasound on the proliferation and adhesion of human adipose-derived mesenchymal stromal cells.
METHODS:  The human adipose-derived mesenchymal stromal cells of the passage 3 were cultured for 24, 48 and 72 hours. The first, second and third times of stimulation were performed with 0, 10, 30, and 50 mW/cm2 low-intensity pulsed ultrasound intensity, and each stimulation time was 5 minutes. After the stimulation, the cells were placed in the incubator for 24 hours. CCK-8 test was used to screen the best stimulation conditions. The cells stimulated twice with 30 mW/cm2 were selected as the experimental group for follow-up experiments, and 0 mW/cm2 for the control group. Flow cytometry was used to detect cell surface markers. Immunohistochemistry was used to observe the expression of proliferation related antigen Ki-67 and focal adhesion kinase FAK. RT-PCR was used to detect the expression of Cyclin D1 and c-myc. Transcriptome sequencing was used to detect the changes of transcription genes. 
RESULTS AND CONCLUSION: (1) 30 mW/cm2 continuous stimulation twice was the best condition to promote the proliferation of human adipose-derived mesenchymal stem cells. (2) Compared with the control group, cell surface markers remained unchanged after 30 mW/cm2 low-intensity pulsed ultrasound stimulation, and the expression of proliferation-related genes CyclinD1, c-myc and focal adhesion kinase FAK were up-regulated. (3) Transcriptome sequencing results showed that there were significant differences in 27 genes, among which 15 genes were up-regulated and 12 genes were down-regulated. (4) It is concluded that low-intensity pulsed ultrasound promotes human adipose-derived mesenchymal stromal cells proliferation in undifferentiated cells by upregulating the expression of CyclinD1, c-myc and FAK, accompanied by regulation of multiple transcriptome genes. 

Key words: stem cells, adipose-derived mesenchymal stem cells, ultrasound, low-intensity pulsed ultrasound, cell proliferation, cell adhesion, gene

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