Peptide‐modified, hyaluronic acid‐based hydrogels as a 3D culture platform for neural stem/progenitor cell engineering

Abstract

Neural stem/progenitor cell (NS/PC)‐based therapies have shown exciting potential for regeneration of the central nervous system (CNS) and NS/PC cultures represent an important resource for disease modeling and drug screening. However, significant challenges limiting clinical translation remain, such as generating large numbers of cells required for model cultures or transplantation, maintaining physiologically representative phenotypes ex vivo and directing NS/PC differentiation into specific fates. Here, we report that culture of human NS/PCs in 3D, hyaluronic acid (HA)‐rich biomaterial microenvironments increased differentiation toward oligodendrocytes and neurons over 2D cultures on laminin‐coated glass. Moreover, NS/PCs in 3D culture exhibited a significant reduction in differentiation into reactive astrocytes. Many NS/PC‐derived neurons in 3D, HA‐based hydrogels expressed synaptophysin, indicating synapse formation, and displayed electrophysiological characteristics of immature neurons. While inclusion of integrin‐binding, RGD peptides into hydrogels resulted in a modest increase in numbers of viable NS/PCs, no combination of laminin‐derived, adhesive peptides affected differentiation outcomes. Notably, 3D cultures of differentiating NS/PCs were maintained for at least 70 days in medium with minimal growth factor supplementation. In sum, results demonstrate the use of 3D, HA‐based biomaterials for long‐term expansion and differentiation of NS/PCs toward oligodendroglial and neuronal fates, while inhibiting astroglial fates. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 704–718, 2019.

Hydrogels Derivatized With Cationic Moieties or Functional Peptides as Efficient Supports for Neural Stem Cells

“…Seidlits et al, 2019). The HA based 3-D hydrogel alone shows an increased differentiation of human NSCs into oligodendrocytes and neurons and a decreased formation of reactive astrocytes compared to a 2-D laminin-coated culture (Seidlits et al, 2019). Additionally, the modification with RGD increased the viability of the human NSCs.…”

“…An often used hydrogel system are the HA based hydrogels ( Table 1; Tarus et al, 2016;Adil et al, 2017;Perera et al, 2019; Madl et al, 2017Madl et al, , 2019 FIGURE 2 | Exemplary synthesis of a synthetic hydrogel system and examples for hydrogels bearing cationic moieties and the RGD sequence. Seidlits et al, 2019). The HA based 3-D hydrogel alone shows an increased differentiation of human NSCs into oligodendrocytes and neurons and a decreased formation of reactive astrocytes compared to a 2-D laminin-coated culture (Seidlits et al, 2019).…”

“…Initially, numerous studies examined the ECM as modulating agents in two-dimensional (2-D) in vitro cell culture systems. Recently, the notion emerged that the three-dimensional (3-D) organization of the ECM exerts specific effects (Duval et al, 2017;Seidlits et al, 2019). In this perspective, a novel aim consisted of finding an appropriate 3-D scaffold for cultivating cells in what is considered a more natural environment.…”

Chemical cross-linking methods for cell encapsulation in hydrogels

“…3D culture of H9 human embryonic stem cells was performed in these gels. [208] Baldwin and Kiick showed that the succinimide thioether linkage resulting from the Michael addition of thiols to maleimides could undergo retro and exchange reactions at physiological pH and temperature. [209] It is possible to stabilize the thioether linkage by opening the succinimide ring.…”

3D Hyaluronic Acid Hydrogels for Modeling Oligodendrocyte Progenitor Cell Behavior as a Function of Matrix Stiffness

“…[7][8][9] OPCs and their precursors (neural stem cells) have been studied in polyethylene glycol (PEG) hydrogels 10 as well as collagen, agarose, chitosan, and fibrin hydrogels. 11,12 Hyaluronic acid (HA) is an excellent candidate for engineering brain-like extracellular matrix (ECM) microenvironments 13,14 due to its abundance in the CNS [15][16][17] and its anti-inflammatory role in brain and spinal cord injuries. 18 Hydrogel cell culture models enable interrogation of the roles that extracellular cues such as matrix stiffness play in regulating salient cell behaviors.…”