1. Synergistic effect of co-immobilized FGF-2 and vitronectin-derived peptide on feeder-free expansion of induced pluripotent stem cells.
- Author
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Sohi, Alireza Naderi, Naderi-Manesh, Hossein, Soleimani, Masoud, Yasaghi, Elaheh Roshani, Manjili, Hamidreza Kheiri, Tavaddod, Sharareh, and Nojehdehi, Shahrzad
- Subjects
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PLURIPOTENT stem cells , *VITRONECTIN , *CELL adhesion molecules , *CHITOSAN , *X-ray photoelectron spectra - Abstract
Abstract Expansion of human induced pluripotent stem cells (h-iPSCs) on mouse derived feeder layers or murine cells secretions such as Matrigel hamper their clinical applications. Alternative methods have introduced novel substrates as stem cell niches or/and optimized combinations of humanized soluble factors as fully defined mediums. Accordingly vitronectin as a main part of ECM have been commercialized significantly as a stem cell niche-forming substrate. In this work, we used a functional peptide derived from vitronectin (VTN) and co-immobilized it with FGF-2 (as an indisputable ingredient of defined culture mediums) on chitosan film surface. After chemical and physical characterization of the pristine chitosan surface as well as ones modified by VTN or/and FGF-2, h-iPS cells were cultured on them at the xeno/feeder-free conditions. Our results demonstrated that co-immobilization of these two biomolecules has a synergistic effect on adhesion and clonal growth of h-iPS cells with maintained expression of pluripotency markers in a FGF-2 density-dependent manner. This is the first report of co-immobilization of an ECM derived molecule and a growth factor for stem cell culture. Graphical abstract Unlabelled Image Highlights • Co-immobilization of an ECM-derived peptide (i.e. vitronectin-derived peptide) and a growth factor (i.e. FGF-2) on the surface (i.e. chitosan film) was accomplished for the first time and confirmed by XPS, fluorometry and ELISA techniques. • The yield of each reaction revealing the surface density of each immobilized biomolecule was estimated which was neglected in the most of immobilization reports. • Feeder/xeno-free culture of human induced pluripotent stem cells (h-iPSCs) on chitosan based surfaces demonstrated that the co-immobilization of vitronectin and FGF-2 has a synergistic effect on attachment, proliferation and colony formation of h-iPS cells with the maintained expression of pluripotency markers in a FGF-2 density-dependent manner. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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