| Recent years, the culture of the neurons has been the hotspot of research. At the same time, the neural stem cell has been dispersed and cultured in vitro successfully. It can provide the theoretic basement for the human being's neural disease, for example, Parkinson syndrome, medulla disease, spinal cord, senile dementia disease. Healthy SD rats were selected as experimental material in this paper. Hippocampus tissue from rat brain was sampled in sterile and the neurons were cultured in vitro. Trypan blue staining, Nissl dyeing, transmission electron microscopy, MTT methods were adopted to judge and check-up the neurons. The results of experiment are followed. 1. Healthy SD rats of 220g-300g were selected as experimental material in this paper. We obtain the separated neurons by the automatically or EDTA-amylopsin digestant method. At the same time, we optimize the cultured condition in vitro to inoculate cells to the board with 4×10~5/mL. The neurons be cultured well by 37℃,5%co2 long for 14-24 days. 2. The effects of different EGF concentration on hippocampal neurons were detected, the results indicated EGF can accelerate the growth of the neuron and the process, specially instinct to the axon growth. The neurons morphologic changes corrected with concentration of EGF linearly——Among them, 20ng/mLEGF is the best concentration. By the morphological observation, it can be seen the more complicated and altitudinal ramiform process. Compared with the contrast group, the difference is extreme distinct in 20ng/mL(P<0.01). It is distinct in 10ng/mL and 100ng/mL EGF(P<0.05), but the difference is not distinct between 10ng/mL and 20ng/mL(P>0.05). 3. There exists a certain extent effect of different Vc concentration on hippocampal neurons, the results show that the comparative high concentration Vc group(200nmol/L) can promote distinctively the cell liability. Compared with the control group, the difference is extreme distince(P<0.01). The fartherly observe the process number, length, area, they are distinct higher to control group(P<0.01). The low concentration of Vc(2 nmol/L, 20nmol/L) was not distinct infection to the hippocampal neurons growth compared with contrast group(P>0.05). However, the high concentration Vc group(2000nmol/L) reduce remarkably cell livability, it can lead to the neurons death. 4. Using two different kinds of basements(Poly-l-lysine ,PLL and rat hail collagen) and their united group to treat to the neurons, the result indicate the time of cell adhibiting the hemline is short in PLL group than the RHC group, but the time of cell casting is earlier. The cell configuration is the similar to the PLL group, the cell dispersal is symmetric, there is scarcely swarmed cell and there is evident luster around the process. It can be seen a lot of bitty tubers out of the process. With the time passed, the most of the neurons grow up the process, which are longer, they can interweave with neurons nearby. The traditional serological cultured method is used, based on it, appending the EGF and the necessary microelement and some growth hormone, we can succeed to culture primary rat hippacampal neurons. More important, we institute the growth system of neurons.
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