Abstract:

BACKGROUND: Cell transplantation for disease treatment is one of interesting topics. To find a better source of seed cells and a more efficient amplification method is the basis for clinical application.
OBJECTIVE: To determine whether the RhoA gene silencing in fetal liver stem cells can optimize liver stem cell cultures.
METHODS: Fetal liver stem cells were in vitro cultured and were transfected by small interfering RNA (siRNA) to knock down the expression of RhoA. Then the cells were randomly divided into three groups. In the group A, fetal liver stem cells were non-treated. In the group B, fetal liver stem cells were transfected with randomly sequenced siRNA. In the group C, fetal liver stem cells were transfected with siRNA. RhoA gene and protein expression prior to and after transfection was detected by RT-PCR and western blot methods, respectively. Cellular proliferation was determined by MTT assay. The cell cycle was analyzed by flow cytometry.
RESULTS AND CONCLUSION: Compared with groups A, B, the expression of RhoA gene and protein in the group C was significantly decreased (P < 0.05), the cell growth speed was significantly increased, cells in the G0/G1 phase of the cell cycle were decreased, and cells in the S phase were increased (P < 0.05). These findings suggest that the RhoA gene silencing can promote the proliferation of fetal liver stem cells and optimize the culture of fetal liver stem cells.