CAJ | 학술논문

目的：优选多聚赖氨酸（ poly－D－lysine，PDL）在大鼠海马神经元原代培养中的最佳工作浓度。方法颈椎脱臼处死孕18 d的Wistar大鼠3只，取出胚胎并分离脑组织。迅速分离海马组织，胰酶消化，吹打并以合适浓度（30万／3．5 cm皿）接种于0．01、0．05、0．1、0．25、0．5、0．75、1、2 mg／mL的PDL工作浓度包被的培养皿中，采用相差显微镜观察神经元形态，同时采用CCK－8试剂盒进行神经元活性检测。结果神经元接种4 h后贴壁，1 d后突起发生，4 d后与周围神经元形成联系，7 d后成熟并形成网状联系。 PDL最佳工作浓度范围位于0．25～0．75 mg／mL，神经元在此浓度范围内生长状态及活性最佳。结论通过实验确定了PDL最佳工作浓度，对于神经元培养有重要意义。
목적：우선다취뢰안산（ poly－D－lysine，PDL）재대서해마신경원원대배양중적최가공작농도。방법경추탈구처사잉18 d적Wistar대서3지，취출배태병분리뇌조직。신속분리해마조직，이매소화，취타병이합괄농도（30만／3．5 cm명）접충우0．01、0．05、0．1、0．25、0．5、0．75、1、2 mg／mL적PDL공작농도포피적배양명중，채용상차현미경관찰신경원형태，동시채용CCK－8시제합진행신경원활성검측。결과신경원접충4 h후첩벽，1 d후돌기발생，4 d후여주위신경원형성련계，7 d후성숙병형성망상련계。 PDL최가공작농도범위위우0．25～0．75 mg／mL，신경원재차농도범위내생장상태급활성최가。결론통과실험학정료PDL최가공작농도，대우신경원배양유중요의의。
Objective To explore working of concentration of PDL used in primary culture of rat hippocampal neurons.Methods 3 pregnant wistar rats were executed by cervical dislocation, the embryos were taken out and the hippocampal tissue was dissected quickly.Then the tissue was digested by trypsin and planted into dishes with proper concentration(300000/3.5cm vessle) which were coated by different PDL solution in different concentration (0.01,0.05,0.1,0.25,0.5,0.75,1,2 mg/mL).The state of the cultured neurons was observed to determine the most suitable concentration of PDL solution in coating dishes.Neurons activity was observed by CCK-8 Kit.Results Most neurons had adhered in 4 h.Protrusion of neurons began to grow in 1 d.The connection between neurons appeared in 4 d.Neurons matured and the network-connection was set up in 7 d.The best working concentration of PDL is between 0.25 and 0.75 mg/mL, neurons grew well and activity was optimum during this concentration.Conclusion The working concentration of PDL is important for the hippocampal neuron culture and this work is worth being applied.