Cell-selective adhesion short peptides for enhancing cell culture on scaffold

Jun 29, 2022, 11:20 AM
10m
Room: S3 A

Room: S3 A

Speaker

Fujimoto, Akiyo (Nagoya University/Graduate School of Pharmaceutical Sciences)

Description

"[Introduction] Extracellular matrix (ECM) plays a critical role in the control of cell adhesion and growth as in vivo scaffold material, therefore the design of ECM-like or ECM-mimicking scaffold has been one of the powerful concepts to provide a successful culture platform for tissue engineering. However, by their molecular size, it has been a challenge to obtain highly purified ECM molecules. Moreover, it is also extremely costly to obtain ECM molecules to be feasibly used for scaffold material. As a result, such ECM-molecular which can be easily obtained and used for scaffold coating has been limited to several bulk-produced proteins, such as collagens. From the aspect of obtaining pure and highly designable functionalization performance, peptides are advantageous with their synthesis and purification feasibility and their designability for effective surface modification applications. Our group has been investigating short peptides, 3-mer peptides, which can control not only cell adhesion but also “cell-selective adhesion”, to design ECM-mimicking surfaces on cell culture scaffolds [1,2]. The enhancement of cell-selective adhesion is expected to improve the regeneration process on the surface of implantable medical devices because they can provide a material surface to be attractive for “objective cells for treatment”, and be inhibitory for “non-objective cells that disturb treatment”. In this research, we tried to screen novel peptide sequences and combinations to enhance such cell-selective adhesion using peptide array-based direct cell assay [3].
[Methods] On the cellulose membrane, a peptide array was designed using Fmoc peptide synthesis. On the solid-bound peptide array, target cells (endothelial cells, fibroblasts, neural cells, smooth muscle cells) were seeded directly, and their cell adhesion rates were evaluated by a plate reader. We designed various short peptide libraries, containing control RGD cell adhesion peptides, and combined their sequences with various amino acid linker sequences.
[Results and conclusions] By the combinatorial direct screening on peptide array, we found that several short 3-mer peptides can be found to control the cell-selective adhesions to relatively discriminate objective cells and non-objective cells. Moreover, from the data, we found that the short peptide-based cell adhesion effect can be greatly influenced by the linker motifs, the neighboring sequence which provides a physicochemical effect for better or worse cell-peptide interaction. Our results indicate the effective performances of short peptides and their effective surface modification methods to provide a cell-selective effect on cell culture scaffolds. Moreover, our data also suggest that it is important to control the cell-surface interaction mechanism which affects apart from the integrin-mediated adhesion.

  1. Kanie, K., et al. Biotech. Bioeng. 109(7), 1808-1816 (2012)
  2. Kanie, K., et al. Materials 9(9), 730 (2016)
  3. Kato R., et al. J. Biosch. Bioeng. 101(6), 485-95 (2006)"

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