Recombinant self‐assembling 16‐residue peptide nanofiber scaffolds for neuronal axonal outgrowth |
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Authors: | Peng Liang Jinsheng Xiong Liwei Zhao Ye Xu Jiaxin Zhao Qing Liu |
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Institution: | 1. Department of Neurosurgery, Affiliated Tumor Hospital, Third Clinical College, Harbin Medical University, Harbin, China;2. Dana‐Farber Cancer Institute, Harvard Medical University, Boston, MA, USA |
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Abstract: | Self‐assembling peptides are considered a good biological scaffold for the repair of injured nervous system. In order to set up a stable system to produce the peptides at low cost, we used a gene recombinant expression method. The sequence of the peptide was devised to facilitate neural cell attachment and growth. The nucleotide sequence of the self‐assembling peptide was designed, artificially synthesized, and inserted into the fusion protein vector pTYB2. After being transformed and expressed in Escherichia coli BL‐21 (DE3) by means of the fusion protein, the soluble 16‐residue peptide (named RAE16) was obtained by one‐step chitin affinity chromatography. During cell culture, bone marrow stromal cells were fully embedded in the 3D environment of the peptide scaffolds. The MTT (3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide) test indicated that bone marrow stromal cells cultured in RAE16 had the highest survival rate with the absorbance value of 0.7 at 7 days. Moreover, the cortical neural axons in the RAE16 group were longer (118.36 ± 7.04 μm) than in the other groups (p < 0.01). The recombinant peptide nanofiber scaffolds we designed provide a promising cell culture system for general molecular and cell biology studies and are useful as well for neural regeneration studies. |
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Keywords: | Axonal outgrowth Neuron Protein expression Self‐assembled nanofiber scaffold Tissue engineering |
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