The Effect of miR-210 Up-regulation on Proliferation and Survival of Mouse Bone Marrow Derived Mesenchymal Stem Cell

Abstract

Background: Bone marrow derived mesenchymal stem cells (MSCs) are a population of multipotent progenitors which have the capacity of proliferation and differentiation into mesenchymal lineage cells. Hypoxia could promote the proliferation of MSCs. Micro-RNAs are endogenous RNAs that can play an important role in some processes such as proliferation and differentiation. MiR-210 could help for better proliferation of MSCs since this miRNA could activate HIF pathway. In current study we investigated if MSCs can preserve their differentiation and proliferation ability under normoxic conditions by upregulation of miR-210.

Materials and methods: MSCs isolated from C57 BL/6 mice by flushing it's femurs into the cell culture media. After 72 hours, MSCs which are plastic adherent cells were attached to the flask and non-adherent cells were removed. Subsequently, MSCs induced to differentiate into osteocytes and adipocytes with specific differentiation media in order to confirm their identity and multipotency. Then miR-210 was inserted in Lentiviruse vectors and affected MSCs. In each passage, the number and viability of cells were evaluated.

Results: Comparison between miR-210 infected MSCs with control cells showed that miR-210 has ability to increase proliferation of MSCs significantly.

Conclusion: We showed that miR-210 has ability to induce proliferation of MSCs without any negative effect on their differentiation abilities. Further studies are needed for evaluation of probable effects of miR-210 mechanisms on MSCs proliferation.

Keywords: HIF-1α; Mesenchymal stem cells; MiR-210.

Fig. 1

The map of plasmid used in this study

Fig. 2

Mesenchymal stem cells 96h after isolation (a), Mesenchymal stem cells 14 days after isolation-3th passage (b)

Fig. 3

Oil Red O staining, intracellular lipid accumulation were stained bright red in adipocytes at day 14 (a), Alizarin Red S staining, calcium deposition were stained bright orange-red in osteocytes at day 21 (b)

Fig. 4

The GFP positive mesenchymal stem cells that infected with vectors without fluorescent microscope (a) and with fluorescent microscope (b)

Fig. 5

Cell number (cell/ml). The control groups are presented in black, cells without miR-210 in white and with miR-210 in gray. *: P value less than 0.01 was considered as statistically significant (P < 0.01).