Designed peptide-grafted hydrogels for human pluripotent stem cell culture and differentiation

Abstract

Human pluripotent stem cells (hPSCs) have the ability to differentiate into cells derived from three germ layers and are an attractive cell source for cell therapy in regenerative medicine. However,...

Design of dual peptide-conjugated hydrogels for proliferation and differentiation of human pluripotent stem cells

“…Pluripotent protein (Oct3/4, Nanog, Sox-2 and SSEA-4) expression in hPSCs was examined by immunohistochemical labelling, following a conventional procedure, using specific antibodies for the pluripotent proteins [ 8 , 28 , 32 , 33 , [35] , [36] , [37] , [38] ]. The labelled cells were assessed with a fluorescence microscope (Eclipse Ti–U, Nikon Instruments, Tokyo, Japan).…”

“…Unlike conventional cells such as cancer cells or primary cells extracted from tissues, hPSCs cannot, in general, proliferate on conventional dishes made of polystyrene. hPSCs are extensively cultured on extracellular matrix (ECM) proteins, for example, in recombinant vitronectin (rVN)-coated or laminin 521 (LN521)-coated dishes [ [8] , [9] , [10] , [11] , [12] ]. Recently, peptide-conjugated materials have been prepared for hPSC proliferation by several researchers [ 8 , [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] ].…”

“…hPSCs are extensively cultured on extracellular matrix (ECM) proteins, for example, in recombinant vitronectin (rVN)-coated or laminin 521 (LN521)-coated dishes [ [8] , [9] , [10] , [11] , [12] ]. Recently, peptide-conjugated materials have been prepared for hPSC proliferation by several researchers [ 8 , [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] ]. Peptide-conjugated cell culture bags, dishes or hydrogels, which can be made in completely xeno-free conditions, are promising biomaterials for hPSC culture and differentiation.…”

“…The peptides are typically derived from vitronectin or laminin for most peptide-conjugated surfaces for hPSC culture [ 8 , [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] ]. hPSCs can bind to vitronectin or laminin via integrin receptors of hPSCs, i.e., αVβ5 or α6β1, respectively, and the binding of hPSCs via integrin αVβ5 or α6β1 can maintain the pluripotency of hPSCs.…”

“…In our previous studies [ 8 , 19 , [26] , [27] , [28] ], we prepared single peptide-conjugated hydrogels using 1000 μg/mL peptide solution for hPSC proliferation and differentiation. Other investigators [ 17 , 19 ] who prepared peptide-conjugated biomaterials also used a relatively high concentration of peptide (>1000 μg/mL) and only a single peptide-conjugated surface or single ECM-derived peptides-conjugated surface.…”