Human embryonic stem cell dispersion in electrospun PCL fiber scaffolds by coating with laminin‐521 and E‐cadherin‐Fc

Abstract

Advances in human pluripotent cell cultivation and differentiation protocols have led to production of stem cell-derived progenitors as a promising cell source for replacement therapy. Three-dimensional (3-D) culture is a better mimic of the natural niche for stem cells and is widely used for disease modeling. Here, we describe a nonaggregate culture system of human embryonic stem cells inside electrospun polycaprolactone (PCL) fiber scaffolds combined with defined extracellular proteins naturally occurring in the stem cell niche. PCL fiber scaffolds coated with recombinant human laminin-521 readily supported initial stem cell attachment and growth from a single-cell suspension. The combination of recombinant E-cadherin-Fc and laminin-521 further improved cell dispersion rendering a uniform cell population. Finally, we showed that the cells cultured in E-cadherin-Fc- and laminin-521-coated PCL scaffolds could differentiate into all three germ layers. Importantly, we provided a chemically defined 3-D system in which pluripotent stem cells grown and differentiated avoiding the formation of cell aggregates. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1226-1236, 2018.

Laminin as a Potent Substrate for Large-Scale Expansion of Human Induced Pluripotent Stem Cells in a Closed Cell Expansion System

“…Microcarriers can be used in suspension bioreactor systems for PSC expansion [36–38]. Using an electrospun polycaprolactone fiber system, Leino et al [36] expanded PSCs in a 3D environment. The authors observed a homogeneous culture environment and the cells maintained pluripotency.…”

“…However, the cells failed to proliferate when low seeding density was used and could not be detached from the system without compromising cell viability due to the porosity of the fibers. Though these systems maintain the stemness of the cells, they exert the negative effect of bead agitation on cell yield, including increased risk of cell detachment [9, 32, 36]. However, lower agitation rates compromise nutrient exchange, resulting in larger aggregates that are difficult to dissociate.…”

“…These coatings have previously been utilized to increase the reliability and reproducibility of cell expansion and differentiation protocols [22, 32, 40, 42, 43]. Both coatings (LN and VN) allowed cells to be expanded, to self-renew with a normal karyotype, and to maintain pluripotency markers; furthermore, they allowed the formation of teratomas containing cells from three germ layers [22, 32, 36, 40, 44]. Although VN could substitute for Matrigel™, Rowland et al [45] demonstrated that Matrigel™ had a slightly better performance in the adhesion and proliferation of cells.…”

Electrospun Poly(ε‐Caprolactone)/Silk Fibroin Coaxial Core‐Sheath Nanofibers Applied to Scaffolds and Drug Carriers

“…The contact angle is widely carried out to measure the fabricated materials hydrophilicity, since it implies the inclination of cell adhesion and proliferation of the material. PCL is inherently hydrophobic thus was usually modified with hydrophilic materials to improve its hydrophilicity, such as collagen, gelatin, laminin, and chitosan [33][34][35][36]. Herein, we found that the hydrophilicity of PCL was greatly increased after it was encapsulated within SF sheath.…”

Extracellular Matrix Coatings on Cardiovascular Materials—A Review

“…ECMbased scaffolds provide a biological matrix with environmental cues that can support the formation of proper vascular tissue. When the scaffold is coated with recombinant human laminin fibers, it supports the attachment and growth of naive stem cells in single-cell suspension [30]. A fibrin-coated pericardial ECM can be used as a material for cardiovascular surgery [31].…”