| Bone marrow stoma not only provides micro surrounding for proliferation and differentiation of hematopoietic cells, but also exist a subset of non-hematopoietic cells referred as mesenchymal stem cells (MSCs). These cells can transdifferentiate into multiple cell lineages including osteoblasts, chondrocytes, adipocytes, tenocytes and myotubes. In given conditions, MSCs can also differentiate into lung cells, cardiac cells and neural cells. MSCs exist within bone marrow as well as umbilical cord blood, peripheral blood and fat tissues et al. The bone marrow is the most important source.The multipotential of bone MSCs, their easy isolation, culture, proliferation and being able to auto-transplant make them ideal engineering cells in cell and gene therapies for central nervous system diseases. MSCs can differentiate into neuron-like cells by using anti-oxidizer in vitro, but the neuron-like cells cannot live for a long time. Gangliosides (GM1) have been shown to re-establish functional recovery of central nervous system that has suffered damage. The basic mechanism for this effect is the phenomenon of neuroplasticity. While weather GM1 has the protection effect on MSCs-derived neuron-like cells has not been reported.Objective:To investigate the feasibility of neuronal induction of bone MSCs from adult rats, and the effects of GM1 on MSC-derived neuron-like cells providing experimental basis for MSCs apply in neuroscience field.Materials and methods:The rat MSCs were isolated primarily from the femurs and tibias of the albino Sprague-Dawley adult rats and maintained in DMEM supplemented with 10% fetal bovine serum (FBS). After 5 days of incubation at 37癈, in 5%CC?, nonadherent cells were removed from the cultures by replacing midium. When the cultures reached confluency, the cells were lifted by incubation with 0.25% trypsin for 2 or 3 minutes at room temperature. MSCs were purified and proliferated by passage culture. At passage 3 rd to 6 th the MSCs were planted into plastic petridishes for induction experiment. MSCs were first cultured with basic fibroblast growth factor (bFGF) for 24 hours for cell proliferation. Then the rat MSCs in experimental group were induced by butylated hydroxyanisole (BHA) and dimethyl sulfoxide (DMSO) for 5 hours. In control group, rMSCs were cultured without any induction medium. Immunocytochemistry were used to identify cell types at different time.After induction for 5 hours the MSCs-derived cells were separated into 6 groups. The induction medium of the 3 GM1 groups was replaced by DMEM containing 3 different levels of GM1 for 48 hours. In DMEM group, GMl-free DMEM was used. In serum group, use DMEM with 10%FBS to replace the induction medium. In induction group the induction medium were maintained. Immunocytochemistry were used to identify cell types at 24 hours and 48 hours after replacing medium.Results:1. The adult rat MSCs can be cultured and proliferated in vitro. The rat MSCs (rMSCs) proliferated twice in 7 days.2. Within 1 hour after induction, some of the rMSCs assumed neuronal morphology. After induction for 5 hours, a large number of responsive cells exhibited typical neuronal morphology: clusters of rMSCs-derived neurons of varying morphologies form complex network.3. The rMSCs in the control group (uninduced rMSCs) expressed nestin and NSE protein, 1.8% 0.8% and 5.6% 2.9% respectively. At 1 hour and 3 hours after induction of rMSCs in the experimental group, the cells expressing nestin reached 40.2% 3.3% and 27.0% + 3.0% respectively. There are significant difference between the two time points (P<0.05), and also between the experimental group and the control group (PO.05). After induction for 5 hours, the cells expressing NSE protein reached 78.8% 3.0%, it is greatly different from the control group (P<0.05). The cells after induction for 5 hours expressing GFAP protein reached 8.6% 1 .8%.4. The induced cells without replacing induction medium (in induction group) were dead or because of apoptosis after 24 hours. The...
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