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

机译:大鼠皮质神经元初级培养中层蛋白-111衍生肽 - 壳聚糖基质的生物活性

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摘要

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 alpha 1 chain 27192730) and C16 (KAFDITYVRLKF, laminin gamma 1 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.
机译:细胞粘合剂生物材料已用于培养中的各种细胞,特别是对于神经元的原代培养物。在这里,我们将层蛋白-111及其与壳聚糖基质(CHTMS)缀合的活性肽用于大鼠皮质神经元的原代培养。聚-D-赖氨酸底物上的层粘连蛋白-111促进了神经元细胞附着和分化。使用肽-CHTM构建体检查六种活性层蛋白-111衍生肽的生物活性。当Syndecan结合肽,AG73(RKRRQVQLSIRT,小鼠层层蛋白α1链27192730)和C16(KafdityVrlkf,Lamininγ1150)与壳聚糖缀合时,Ag73-CHTM和C16-CHTM显示有效的神经元细胞附着活性通过初级培养的大鼠皮质神经元促进轴突延伸。然而,剩余的肽(包括整联蛋白结合肽)在与CHTM缀合时没有显示出活动。 AG73-CHTM和C16-CHTM还支持Neuron存活率在无胶质喂食器层的无血清培养基中至少4周。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物,并且有可能用作中枢神经系统中的组织工程的功能生物材料。

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