Varon S
Neurochem Res. 1979 Apr;4(2):155-73. doi: 10.1007/BF00964141.
Polylysine-coated culture surfaces are strongly adhesive for neural cells, restrict locomotion on nonneuronal elements, but do not inhibit neurite elongation. In the present study, culture dishes were pre-treated with poly-D-lysine (PDL) at various concentrations, seed with dissociates from 8-day chick embryo dorsal root ganglia, and incubated under conditions that normally support both neuronal survival and nonneuronal proliferation. Pretreatment with low (0.1 mg/ml) PDL concentrations had no effect on neuronal survival and neuritic growth, but entirely prevented an increase in ganglionic nonneurons, yielding a numericallly stable culture greatly enriched in neurons. Higher PDL concentrations caused increasing losses in both cell classes. The 50% levels of cell loss were achieved at about the same PDL dose, but earlier for neurons that nonneurons and still with no impairment of neuritic growth from the surviving neurons. A procedure was developed to compare acid-soluble and acid-precipitable accumulation of radioactivity under 1-hr pulses of [3H]uridine, which was applicable even to poorly attached cells. The cytotoxic effects of higher PDL pretreatments was revealed as early as 6 hr after seeding by 2- to 4-fold lower radioaccumulation. The data are discussed in terms of possible regulations of cell permeability and metabolism by adhesive interactions between cells and their substratum, or other cells.
聚赖氨酸包被的培养表面对神经细胞具有很强的黏附性,限制其在非神经元成分上的移动,但不抑制神经突的伸长。在本研究中,培养皿用不同浓度的聚-D-赖氨酸(PDL)进行预处理,接种8日龄鸡胚背根神经节的解离细胞,并在通常支持神经元存活和非神经元增殖的条件下孵育。低浓度(0.1 mg/ml)的PDL预处理对神经元存活和神经突生长没有影响,但完全阻止了神经节中非神经元数量的增加,从而得到神经元数量稳定且大量富集的培养物。较高浓度的PDL导致两类细胞的损失增加。细胞损失50%的水平在大约相同的PDL剂量下达到,但神经元比非神经元更早出现损失,且存活神经元的神经突生长未受损害。开发了一种程序来比较在[3H]尿苷1小时脉冲下放射性的酸溶性和酸沉淀性积累,该程序甚至适用于贴壁不良的细胞。较高PDL预处理的细胞毒性作用在接种后6小时就表现为放射性积累降低2至4倍。从细胞与其基质或其他细胞之间的黏附相互作用对细胞通透性和代谢的可能调节方面对数据进行了讨论。