Jucker M, Kleinman H K, Ingram D K
Laboratory of Cellular and Molecular Biology, National Institute on Aging, National Institutes of Health, Baltimore, Maryland 21224.
J Neurosci Res. 1991 Apr;28(4):507-17. doi: 10.1002/jnr.490280407.
Responses of rat embryonic septal cells to reconstituted basement membrane, laminin, and laminin A chain-derived synthetic peptides were studied in culture. Dissociated fetal E16/17 septal cells were grown for three days on differently coated plastic substrata. Reconstituted basement membrane (Matrigel), laminin, and a 19-amino acid synthetic peptide CSRARKQAASIKVAVSADR-NH2 (PA22-2) from the laminin A chain sequence mediated cell-substratum adhesion and promoted neurite outgrowth. In contrast, cells did not attach to or form processes on uncoated plastic or on plastic substrata coated with synthetic, laminin-derived control peptides. Polyethylenimine (PEI) supported the adhesion and survival of fetal septal cells; however, when laminin was added to the medium during cell plating or 18 hr afterward, a dose-dependent increase was observed in neurite outgrowth of cells attached to this substratum. Cells grown for 6 days on PEI in the presence of laminin showed a determined increase in the number of cholinergic neurons as marked by acetylcholinesterase staining. These data suggest that the subpopulation of cholinergic septal neurons present in the septal cells studied here were also responding to laminin. The results of this in vitro study suggest potential uses for basement membrane, laminin, or synthetic peptides, such as PA22-2, in fetal septal grafts to enhance regeneration in the damaged septo-hippocampal system.
在培养过程中研究了大鼠胚胎中隔细胞对重组基底膜、层粘连蛋白和层粘连蛋白A链衍生的合成肽的反应。将解离的胚胎E16/17期胎儿中隔细胞在不同包被的塑料基质上培养三天。重组基底膜(基质胶)、层粘连蛋白以及来自层粘连蛋白A链序列的19个氨基酸的合成肽CSRARKQAASIKVAVSADR-NH2(PA22-2)介导细胞与基质的黏附并促进神经突生长。相比之下,细胞在未包被的塑料或包被有合成的、层粘连蛋白衍生的对照肽的塑料基质上不黏附也不形成突起。聚乙烯亚胺(PEI)支持胎儿中隔细胞的黏附和存活;然而,当在细胞接种时或之后18小时向培养基中添加层粘连蛋白时,观察到黏附于该基质的细胞的神经突生长呈剂量依赖性增加。在层粘连蛋白存在的情况下在PEI上生长6天的细胞,经乙酰胆碱酯酶染色显示胆碱能神经元数量有明显增加。这些数据表明,此处研究的中隔细胞中存在的胆碱能中隔神经元亚群也对层粘连蛋白有反应。这项体外研究的结果表明,基底膜、层粘连蛋白或合成肽(如PA22-2)在胎儿中隔移植中具有潜在用途,可增强受损中隔-海马系统的再生。