Stoppini L, Buchs P A, Muller D
Department of Pharmacology, Centre Médical Universitaire, Geneva, Switzerland.
J Neurosci Methods. 1991 Apr;37(2):173-82. doi: 10.1016/0165-0270(91)90128-m.
Hippocampal slices prepared from 2-23-day-old neonates were maintained in culture at the interface between air and a culture medium. They were placed on a sterile, transparent and porous membrane and kept in petri dishes in an incubator. No plasma clot or roller drum were used. This method yields thin slices which remain 1-4 cell layers thick and are characterized by a well preserved organotypic organization. Pyramidal neurons labelled by extra- and intracellular application of horse radish peroxidase resemble by the organization and complexity of their dendritic processes those observed in situ at a comparable developmental stage. Excitatory and inhibitory synaptic potentials can easily be analysed using extra- or intracellular recording techniques. After a few days in culture, long-term potentiation of synaptic responses can reproducibly be induced. Evidence for a sprouting response during the first days in culture or following sections is illustrated. This technique may represent an interesting alternative to roller tube cultures for studies of the developmental changes occurring during the first days or weeks in culture.
从2至23日龄新生儿制备的海马切片在空气与培养基的界面处进行培养。它们被放置在无菌、透明且多孔的膜上，并保存在培养箱中的培养皿中。未使用血浆凝块或转鼓。这种方法产生的薄片保持1至4个细胞层厚，其特征是具有保存良好的器官型组织结构。通过细胞外和细胞内应用辣根过氧化物酶标记的锥体神经元，其树突过程的组织和复杂性与在可比发育阶段原位观察到的相似。使用细胞外或细胞内记录技术可以轻松分析兴奋性和抑制性突触电位。培养几天后，可重复性地诱导突触反应的长时程增强。展示了培养初期或切片后发芽反应的证据。对于研究培养最初几天或几周内发生的发育变化，该技术可能是转管培养的一个有趣替代方法。
