Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds

Abstract

Human embryonic stem (hES) cells hold promise as an unlimited source of cells for transplantation therapies. However, control of their proliferation and differentiation into complex, viable 3D tissues is challenging. Here we examine the use of biodegradable polymer scaffolds for promoting hES cell growth and differentiation and formation of 3D structures. We show that complex structures with features of various committed embryonic tissues can be generated, in vitro, by using early differentiating hES cells and further inducing their differentiation in a supportive 3D environment such as poly(lactic-co-glycolic acid)͞poly(L-lactic acid) polymer scaffolds. We found that hES cell differentiation and organization can be influenced by the scaffold and directed by growth factors such as retinoic acid, transforming growth factor ␤, activin-A, or insulin-like growth factor. These growth factors induced differentiation into 3D structures with characteristics of developing neural tissues, cartilage, or liver, respectively. In addition, formation of a 3D vessel-like network was observed. When transplanted into severe combined immunodeficient mice, the constructs continue to express specific human proteins in defined differentiated structures and appear to recruit and anastamose with the host vasculature. This approach provides a unique culture system for addressing questions in cell and developmental biology, and provides a potential mechanism for creating viable human tissue structures for therapeutic applications.

Synthetic nanostructures inducing differentiation of human mesenchymal stem cells into neuronal lineage

“…The suppression might be due to the interference of nanotopography-induced signal with the RA signaling. Although the concentration of 1μM of RA has been shown to be sufficient in inducing neuronal differentiation in embryonic stem cells [27] and neuronal stem cells [28], a concentration of 30μM RA is necessary to achieve the neuronal differentiation of hMSCs [29]. A higher concentration of the RA might be needed to overcome the lineage restriction of MSCs to become neuronal-like cells.…”

Oriented, Multimeric Biointerfaces of the L1 Cell Adhesion Molecule: An Approach to Enhance Neuronal and Neural Stem Cell Functions on 2-D and 3-D Polymer Substrates

“…This suggests cooperative enhancement of L1 signaling on subcellular scales of substrate geometry, a phenomenon that deserves further mechanistic analysis. Scaffold topography has been shown to influence and enhance cellular responses, [65][66][67] as the 3-D architecture better emulates in vivo conditions compared to 2-D thin films.…”

Endochondral bone tissue engineering using embryonic stem cells

“…Besides efforts to increase bone formation by MSCs in vivo, we also explore ES cells (ESCs) (6)(7)(8) as a potential source for bone tissue engineering. ESCs are capable of indefinite undifferentiated proliferation in vitro and can provide an unlimited supply of cells, which can be differentiated into various cell types (9).…”