Manthorpe M, Luyten W, Longo F M, Varon S
Brain Res. 1983 May 9;267(1):57-66. doi: 10.1016/0006-8993(83)91039-9.
Dissociated 4-day (stage 23) chick embryo lumbar cord cells were cultured at low or high cell densities for 1 or 5 days in the presence or absence of added spinal neuronotrophic factor (supplied as RN22 Schwannoma conditioned medium, RCM). In low density, 1-day cultures neuronal survival was dependent on added RCM whereas by 5 days no neurons survived, even in the presence of RCM. In high density 1-day cultures a substantial neuronal population could survive even without added RCM and a large proportion of this neuronal population would survive for 5 days. When conditioned media from high density lumbar cord cultures was supplied to low density unsupplemented cultures, a similar level of 5-day neuronal survival resulted. However, no neurons survived in RCM-supplemented 5-day high density cultures, indicating the presence in RCM of a material toxic for the neurons. Both the RCM and the high density lumbar culture-conditioned medium supported considerable choline acetyltransferase activity indicating the presence within these cultures of motoneurons.
将解离的4日龄(第23阶段)鸡胚腰髓细胞在添加或不添加脊髓神经营养因子(以RN22雪旺瘤条件培养基,RCM提供)的情况下,以低密度或高密度培养1天或5天。在低密度下,1天的培养物中神经元存活依赖于添加的RCM,而到5天时,即使存在RCM也没有神经元存活。在高密度1天的培养物中,即使不添加RCM,大量神经元群体也能存活,并且该神经元群体的很大一部分能存活5天。当将高密度腰髓培养物的条件培养基提供给未添加补充剂的低密度培养物时,产生了类似水平的5天神经元存活。然而,在添加RCM的5天高密度培养物中没有神经元存活,表明RCM中存在对神经元有毒的物质。RCM和高密度腰髓培养条件培养基都支持相当可观的胆碱乙酰转移酶活性,表明这些培养物中存在运动神经元。