Effects of hyperbaric oxygen intervention on the culture and proliferation of bone marrow mesenchymal stem cells in mice labeled by green fluorescent protein

Abstract

Objective To investigate the effects of hyperbaric oxygen (HBO) on the culture and proliferation of bone marrow mesenchymal stem cells (MSCs). Methods MSCs were derived from mice labeled by green fluorescent protein, isolated from whole bone marrow cells by adherent culture method. MSCs of the third generation were induced into neuron-like cells by using galactose-cerebroside and gliocyte-derived nerve growth factor coupled with daily HBO intervention. The expression of neuron specific enolase (NSE) was detected with immunofluorescence, and cell growth curve was delineated. Results MSCs could be ideally isolated by adherent culture method after repeated replacement of culture medium. Following HBO intervention, the exponential stage of MSC growth in the HBO group was advanced and the number of MSCs in the plateau stage was greater than that of the control group. Twenty-four hours after HBO intervention, axons and dendrites could be seen in the HBO group. After forty-eight hours, when comparisons were made between the 2 groups, noticeable changes could be found in MSCs. In the HBO group, processes were found in more than 80% MSCs, with the processes being relatively long and thick. By comparison, in the control group, less than 80% MSCs were detected to have processes, which were short and dispersed. Conclusions The whole bone marrow mesenchymal stem cell adherent culture method had the advantages of simple manipulation and good cell reproductivity, and HBO intervention could accelerate the growth of MSCs. In the induction of the cells supplemented by HBO intervention, large numbers of cells were differentiated with longer processes, indicating that HBO intervention could enhance the proliferation and differentiation of cells from in vitro bone marrow MSCs to neuron-like cells. Key words: Hyperbaric oxygen; Mice labeled by green fluorescent protein; Bone marrow mesenchymal stem cells; In vitro culture