A chemically defined substrate for the expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells

Abstract

Due to the limitation of current pharmacological therapeutic strategies, stem cell therapies have emerged as a viable option for treating many incurable neurological disorders. Specifically, human pluripotent stem cell (hPSC)-derived neural progenitor cells (hNPCs), a multipotent cell population that is capable of near indefinite expansion and subsequent differentiation into the various cell types that comprise the central nervous system (CNS), could provide an unlimited source of cells for such cell-based therapies. However the clinical application of these cells will require (i) defined, xeno-free conditions for their expansion and neuronal differentiation and (ii) scalable culture systems that enable their expansion and neuronal differentiation in numbers sufficient for regenerative medicine and drug screening purposes. Current extracellular matrix protein (ECMP)-based substrates for the culture of hNPCs are expensive, difficult to isolate, subject to batch-to-batch variations, and, therefore, unsuitable for clinical application of hNPCs. Using a high-throughput array-based screening approach, we identified a synthetic polymer, poly(4-vinyl phenol) (P4VP), that supported the long-term proliferation and self-renewal of hNPCs. The hNPCs cultured on P4VP maintained their characteristic morphology, expressed high levels of markers of multipotency, and retained their ability to differentiate into neurons. Such chemically defined substrates will eliminate critical roadblocks for the utilization of hNPCs for human neural regenerative repair, disease modeling, and drug discovery.

Neurite extension and neuronal differentiation of human induced pluripotent stem cell derived neural stem cells on polyethylene glycol hydrogels containing a continuous Young's Modulus gradient

“…Many matrix properties affect cellular response . Even small changes in substrate properties can lead to significant changes in cellular response .…”

Combinatorial extracellular matrix microenvironments promote survival and phenotype of human induced pluripotent stem cell-derived endothelial cells in hypoxia

“…In addition, the microarray platform was utilized to identify synthetic polymers that promote attachment, proliferation and self-renewal among human embryonic stem cell lines [33]. More recently, Brafman et al reported that this platform could be used to examine ECMs that regulate endodermal differentiation in human ESCs [35], as well as identify synthetic polymers that support neuronal differentiation of neural progenitor cells [41]. These aforementioned works primarily employed ECM microarrays to identify the ideal microenvironment niche for cell maintenance and differentiation.…”

Combinatorial Extracellular Matrix Microenvironments for Probing Endothelial Differentiation of Human Pluripotent Stem Cells

“…In addition, the microarray platform was used to select synthetic polymers that improve attachment, proliferation and self-renewal among multiple human ESCs lines over five passages 15 . More recently, it was reported that ECM microarrays were used to study the ECMs that regulate endodermal differentiation in human ESCs 16 , as well as identify synthetic polymers that promote neuronal differentiation of progenitor cells 17 .…”