骨髓基质细胞分泌物促进体外脊髓运动神经元的存活和生长

Bone marrow stromal cells secretory factors promotes survival and growth of spinal cord motoneuron in vitro

目的 体外检测骨髓基质细胞 (Bonemarrowstromalcells ,BMSCs)分泌物对运动神经元的活性作用。方法 体外培养 3 0只昆明小鼠的骨髓细胞 ,利用骨髓基质细胞容易贴附塑料壁的特点 ,纯化骨髓基质细胞并收集其条件培养液 ,将浓缩后的条件培养液按不同的体积百分比浓度加至脊髓运动神经元培养体系中 ,采用MTT法测定培养后 2 4h神经元的活性和相差倒置显微镜计量 ,及培养后 72h、1周的细胞形态学指标。结果 运动神经元的活性变化与条件培养液浓度呈显著的相关性 (r =0 .9589,P <0 .0 0 5) ;贴壁细胞数、细胞直径、突起长度、有突细胞 /总细胞数均随条件培养液浓度的增加而增加。结论 BMSCs在生理状态下合成和分泌有神经营养活性的物质 ,该物质对脊髓运动神经元有维持存活和促进生长的营养作用。

Objective To investigate the effects of bone marrow stromal cells (BMSCs) secretory factors on cultured spinal cord motor neuron. Methods BMSCs was cultured from 30 Kunming mice in vitro. BMSCs were purified due to its adherence to plastic. The conditioned media was collected, centrifuged and concentrated at ice cold condition. Then the preparation was added to motor neuron of cultured mouse spinal cord according to different volume percentage concentration. After 24 hours, the surviving activity of motor neurons was examined using MTT method, and the morphology was observed using phase contrast microscope at 72 hours and 1 week. Results The surviving activity of motor neurons was significantly correlated to concentration of the preparation(r = 0.958 9, P < 0.005). And the increase of cell anchoring, diameter, process length, the ratio of cells with process to gross number of cells was paralleled to the rise of concentration of the preparation. Conclusions BMSCs can synthesize and secrete neurotrophic factors physiologically. BMSCs secretory factors play an important role of promoting the survival and growth of motor neuron.