Influence of glial cell line-derived neurotrophic factors on spinal cord motor neurons in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND: Glial cell line-derived neurotrophic factor (GDNF) ischaracterized by its trophic function on motor neurons, but there is stilllack of quantitative data concerning the influence of different concentra tions of the neurotrophic factor on the growth of in vitro cultured motorneurons. OBJECTIVE: To observe the influence of GDNF on neuronal growth byobserving fetus rat spinal cord motor neurons cultured in vitro. DESIGN: Verifying observation taking in vitro cultured cells as subjects. SETTING: Neurological Department of the Second Hospital Affiliated toHebei Medical College. MATERIALS This experiment was conducted in the laboratory of Neu rological Department, the Second Hospital Affiliated to Hebei Medical College, between January 2001 and September 2002. Adult male and female rats were raised together in the same cage, embryonic rats at 15 days of gestation were obtained for spinal cord separation. METHODS: Ventral spinal tissues were obtained from embryonic rats at 15 days of gestation for prinary in vitro culture. They were divided into four groups according to the density of GDNF, namely 1, 10, 50 and 100 μg/L GDNF groups, while the culture medium in control group did not contain GDNF. Neurons were cultured in 8 wells foreach group, which was repeated for two batches. Then the influence of GDNF on spinal cord motor neu rons was observed from the perspective of cell morphology with MTF method. MAIN OUTCOME MEASURES: The survival rate of motor neurons andthe length of cell processes. RESULTS: ① The length of spinal cord motor neuronal processes It was found obviously longer in GDNF 1 μg/L group, 10 μg/L group, 50 μg/L group and 100 μg/L group than in control group [(107.4±35.406 8,160.5±38.564 9, 450.5±60.640 3, 293.5±67.381 4, 82.8±7.972 5) μm, t=2.610-2.647, P ＜ 0.01]. ② Cell survival rate It was higher in GDNF 1 μg/L group, 10 μg/L group, 50 μg/L group and 100 μ g/L group than in control group [(13.9±0.899 9, 16.1±0.668 0, 20.1±0.667 9, 26.0±0.603 0,10.5±0.782 0) μm, t=2.211-2.312, P ＜ 0.05]. ③ MTT colorimetric analy sis It was obviously higher in GDNF 1 μg/L group, 10 μg/L group, 50 μg/Lgroup and 100 μ g/L group than in control group [(0.350±0.059 8, 0.366 7±0.071 9, 0.381 9±0.063 8, 0.395 3±0.060 5, 0.285 8±0.032 5) μm,t=2.259-2.577, P ＜ 0.05]. CONCLUSION: GDNF of different concentrations exerts different effects on in vitro cultured embryonic spinal cord motor neurons.