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用于神经干细胞培养的重组蜘蛛丝基质。

Recombinant spider silk matrices for neural stem cell cultures.

机构信息

Linnaeus Center in Developmental Biology for Regenerative Medicine (DBRM), Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.

出版信息

Biomaterials. 2012 Nov;33(31):7712-7. doi: 10.1016/j.biomaterials.2012.07.021. Epub 2012 Aug 3.

Abstract

Neural stem cells (NSCs) have the capacity to differentiate into neurons, astrocytes, and oligodendrocytes. Accordingly, NSCs hold great promise in drug screening and treatment of several common diseases. However, a major obstacle in applied stem cell research is the limitation of synthetic matrices for culturing stem cells. The objective of this study was to evaluate the suitability of recombinant spider silk (4RepCT) matrices for growth of NSCs. NSCs isolated from the cerebral cortices of mid-gestation rat embryos were cultured on either 4RepCT matrices or conventional poly-L-ornithine and fibronectin (P + F) coated polystyrene plates. From 48 h of culture, no significant differences in cell proliferation or viability were detected in NSC cultures on 4RepCT compared to control matrices (polystyrene plates coated with P + F). The NSCs retained an undifferentiated state, displaying low or no staining for markers of differentiated cells. Upon stimulation NSCs grown on 4RepCT differentiated efficiently into neuronal and astrocytic cells to virtually the same degree as control cultures, but a slightly less efficient oligodendrocyte differentiation was noted. We suggest that recombinant spider silk matrices provide a functional microenvironment and represent a useful tool for the development of new strategies in neural stem cell research.

摘要

神经干细胞(NSCs)具有分化为神经元、星形胶质细胞和少突胶质细胞的能力。因此,NSCs 在药物筛选和治疗几种常见疾病方面具有巨大的潜力。然而,应用干细胞研究中的一个主要障碍是用于培养干细胞的合成基质的局限性。本研究旨在评估重组蜘蛛丝(4RepCT)基质对 NSCs 生长的适用性。从中孕期大鼠胚胎大脑皮质分离的 NSCs 分别在 4RepCT 基质或传统的多聚-L-鸟氨酸和纤维连接蛋白(P + F)包被聚苯乙烯板上培养。从培养的 48 小时开始,与对照基质(用 P + F 包被的聚苯乙烯板)相比,在 4RepCT 上培养的 NSCs 的细胞增殖或活力没有明显差异。NSCs 保持未分化状态,对分化细胞标志物的染色低或无。在刺激下,在 4RepCT 上生长的 NSCs 有效地分化为神经元和星形胶质细胞,与对照培养物几乎相同程度,但少突胶质细胞的分化效率略低。我们认为,重组蜘蛛丝基质提供了功能性的微环境,是开发神经干细胞研究新策略的有用工具。

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