Simvastatin enhances proliferation and pluripotent gene expression by canine bone marrow-derived mesenchymal stem cells (cBM-MSCs) in vitro

Abstract

Establishing the intervention to enhance proliferation and differentiation potential is crucial for the clinical translation of stem cell-based therapy. In this study, the effects of simvastatin on these regards were explored. Canine bone marrow-derived mesenchymal stem cells (cBM-MSCs) were treated with 4 doses of simvastatin, 0.1, 1, 10, and 100 nM. Simvastatin in low-dose range, 0.1 and 1 nM, enhanced dose-dependent cell proliferation at day 5 and 7. Exploration of the mechanisms revealed that simvastatin in low-dose range dose-dependently upregulated sets of cell cycle regulators, Cyclin D1 and Cyclin D2; proliferation marker, Ki-67; and anti-apoptotic gene; Bcl-2. Interestingly, pluripotent markers, Rex1 and Oct4, were dramatically increased upon the low-dose treatment. Contrastingly, treatment with high-dose simvastatin suppressed the expression of those genes. Thus, the results suggested beneficial effects of simvastatin on cBM-MSCs proliferation and expansion. Further study regarding differentiation potential and underlying mechanisms will accelerate the clinical application of the molecule on veterinary stem cell-based therapy.

Mesenchymal stem cell-based bone tissue engineering for veterinary practice

“…In this regard, strategy to enhance cell isolation yield and proliferation potential of cBM-MSCs is indeed necessitated for improving the potential application of the cells for bone tissue engineering. Our report has recently suggested the beneficial effects of simvastatin in low-dose range, 0.1 and 1 nM, on cBM-MSCs proliferation and pluripotent marker expression in vitro (Nantavisai et al., 2019).…”

Tailored generation of insulin producing cells from canine mesenchymal stem cells derived from bone marrow and adipose tissue

“…In this study, the pancreatic differentiation potential of cBM-MSCs and cAD-MSCs was evaluated to determine the feasibility of IPC formation in vitro and the potential of their clinical application. The cBM-MSCs and cAD-MSCs were isolated, cultured, and expanded using previous published protocols 17 , 18 , 30 , 31 . Their characteristics were similar to those described in previous reports, including fibroblast-like structures, mRNA expressions related to stemness and proliferation, and MSC-related surface marker expression, along with the multilineage differentiation potential toward osteogenic, chondrogenic, and adipogenic lineages 16 – 18 , 31 – 34 .…”

“…Multilineage differentiation potential was assessed regarding osteogenicity, chondrogenicity, and adipogenicity. For osteogenic differentiation, the previously published induction protocol was used 17 , 21 , 103 . Briefly, the cells were seeded onto a 24-well culture plate (Corning, USA) in a concentration of 2.5 × 10 5 cells/well.…”

Tailor-made generation of insulin-producing cells from canine mesenchymal stem cells derived from bone marrow and adipose tissue

“…In this study, the pancreatic differentiation potential of cBM-MSCs and cAD-MSCs was evaluated aiming for determining the feasibility of IPC formation in vitro and the potential of their clinical application. The cBM-MSCs and cAD-MSCs were isolated, cultured, and expanded using previous published protocols [25][26][27][28] . Their characteristics were similar as described in previous reports including fibroblast-like structure, mRNA expression related to stemness and proliferation, MSC-related surface marker expression, and osteogenic differentiation potential [26][27][28][29][30] .…”

“…The cBM-MSCs and cAD-MSCs were isolated, cultured, and expanded using previous published protocols [25][26][27][28] . Their characteristics were similar as described in previous reports including fibroblast-like structure, mRNA expression related to stemness and proliferation, MSC-related surface marker expression, and osteogenic differentiation potential [26][27][28][29][30] . It should be noted that the expression of Cd73 in both MSCs was relatively low as mentioned in previous report 31 .…”