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层粘连蛋白-111 衍生肽-壳聚糖基质在原代培养大鼠皮质神经元中的生物学活性。

Biological activities of laminin-111-derived peptide-chitosan matrices in a primary culture of rat cortical neurons.

机构信息

Department of Applied Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, 192-0392, Japan.

Department of Applied Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, 192-0392, Japan.

出版信息

Arch Biochem Biophys. 2018 Jun 15;648:53-59. doi: 10.1016/j.abb.2018.04.010. Epub 2018 Apr 17.

Abstract

Cell adhesive biomaterials have been used for various cells in culture, especially for primary cultures of neurons. Here we examined laminin-111 and its active peptides conjugated to chitosan matrices (ChtMs) for primary culture of rat cortical neurons. Laminin-111 on poly-d-lysine substrate promoted neuronal cell attachment and differentiation. The biological activity of six active laminin-111-derived peptides was examined using a peptide-ChtM construct. When the syndecan-binding peptides, AG73 (RKRLQVQLSIRT, mouse laminin α1 chain 2719-2730) and C16 (KAFDITYVRLKF, laminin γ1 chain 139-150), were conjugated to chitosan, AG73-ChtM and C16-ChtM showed potent neuronal cell attachment activity and promoted axon extension by primary cultured rat cortical neurons. However, the remaining peptides, including integrin-binding peptides, did not show activity when conjugated to ChtM. AG73-ChtM and C16-ChtM also supported neuron survival for at least 4 weeks in serum-free medium without a glia feeder layer. These data suggest that AG73-ChtM and C16-ChtM are useful for primary cultures of central nervous system neurons and have a potential for use as functional biomaterials for tissue engineering in the central nervous system.

摘要

细胞黏附性生物材料已被用于各种细胞培养,特别是用于原代神经元培养。在此,我们研究了层粘连蛋白-111 及其与壳聚糖基质(ChtM)结合的活性肽,用于原代培养大鼠皮质神经元。多聚-d-赖氨酸基质上的层粘连蛋白-111 促进神经元细胞附着和分化。使用肽-ChtM 构建物研究了六个活性层粘连蛋白-111 衍生肽的生物学活性。当将整联蛋白结合肽 AG73(RKRLQVQLSIRT,鼠层粘连蛋白 α1 链 2719-2730)和 C16(KAFDITYVRLKF,层粘连蛋白 γ1 链 139-150)缀合到壳聚糖上时,AG73-ChtM 和 C16-ChtM 表现出强烈的神经元细胞附着活性,并促进原代培养的大鼠皮质神经元轴突延伸。然而,其余的肽,包括整合素结合肽,当与 ChtM 缀合时没有活性。AG73-ChtM 和 C16-ChtM 还支持神经元在无血清培养基中至少 4 周的存活,而无需胶质细胞饲养层。这些数据表明,AG73-ChtM 和 C16-ChtM 可用于中枢神经系统神经元的原代培养,并且有可能用作中枢神经系统组织工程的功能性生物材料。

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