Mammadov Busra, Guler Mustafa O, Tekinay Ayse B
Institute of Materials Science and Nanotechnology, National Nanotechnology Research Center (UNAM), Bilkent University, Ankara, 06800, Turkey.
Methods Mol Biol. 2014;1202:131-48. doi: 10.1007/7651_2013_35.
Self-assembled peptide nanofibers form three-dimensional networks that are quite similar to fibrous extracellular matrix (ECM) in their physical structure. By incorporating short peptide sequences derived from ECM proteins, these nanofibers provide bioactive platforms for cell culture studies. This protocol provides information about preparation and characterization of self-assembled peptide nanofiber scaffolds, culturing of neural stem cells (NSCs) on these scaffolds, and analysis of cell behavior. As cell behavior analyses, viability and proliferation of NSCs as well as investigation of differentiation by immunocytochemistry, qRT-PCR, western blot, and morphological analysis on ECM mimetic peptide nanofiber scaffolds are described.
自组装肽纳米纤维形成三维网络，其物理结构与纤维状细胞外基质（ECM）非常相似。通过整合源自ECM蛋白的短肽序列，这些纳米纤维为细胞培养研究提供了生物活性平台。本方案提供了有关自组装肽纳米纤维支架的制备和表征、在这些支架上培养神经干细胞（NSCs）以及细胞行为分析的信息。作为细胞行为分析，描述了NSCs在ECM模拟肽纳米纤维支架上的活力和增殖，以及通过免疫细胞化学、qRT-PCR、蛋白质印迹和形态分析对分化的研究。
