Bioorthogonally Cross‐Linked Hyaluronan–Laminin Hydrogels for 3D Neuronal Cell Culture and Biofabrication

Abstract

Laminins (LNs) are key components in the extracellular matrix of neuronal tissues in the developing brain and neural stem cell niches. LN‐presenting hydrogels can provide a biologically relevant matrix for the 3D culture of neurons toward development of advanced tissue models and cell‐based therapies for the treatment of neurological disorders. Biologically derived hydrogels are rich in fragmented LN and are poorly defined concerning composition, which hampers clinical translation. Engineered hydrogels require elaborate and often cytotoxic chemistries for cross‐linking and LN conjugation and provide limited possibilities to tailor the properties of the materials. Here a modular hydrogel system for neural 3D cell cultures, based on hyaluronan and poly(ethylene glycol), that is cross‐linked and functionalized with human recombinant LN‐521 using bioorthogonal copper‐free click chemistry, is shown. Encapsulated human neuroblastoma cells demonstrate high viability and grow into spheroids. Long‐term neuroepithelial stem cells (lt‐NES) cultured in the hydrogels can undergo spontaneous differentiation to neural fate and demonstrate significantly higher viability than cells cultured without LN. The hydrogels further support the structural integrity of 3D bioprinted structures and maintain high viability of bioprinted and syringe extruded lt‐NES, which can facilitate biofabrication and development of cell‐based therapies.

Click chemistry for 3D bioprinting

“…This effectively address the issue of the poor mechanical properties of the pure MHA scaffold which results in poorly printed constructs. For instance, Jury et al , 211 fabricated 3D bioprinting-compatible modular HA-based hydrogel. 211 In the study, full-size laminins (LNs) were successfully conjugated to HA-based hydrogel system via biorthogonal copper-free click reaction and retained for 7 days.…”

“…For instance, Jury et al , 211 fabricated 3D bioprinting-compatible modular HA-based hydrogel. 211 In the study, full-size laminins (LNs) were successfully conjugated to HA-based hydrogel system via biorthogonal copper-free click reaction and retained for 7 days. The human neuroblastoma cells (SH-SY5Y cells) encapsulated in LN-functionalized HA-based hydrogel were well protected and were not affected by lethal shear forces.…”

Cryopreservation of Cell Sheets for Regenerative Therapy: Application of Vitrified Hydrogel Membranes

“…Human laminin-511 supports the stable culture of hESCs and hiPSCs [ 54 ]. Laminin is an important constituent in neuronal tissue and brain [ 55 ]. To create three-dimensional neuronal models with neurons that are similar to those of living organisms, it is useful to incorporate laminin into hydrogels.…”

Enhancing Stem Cell-Based Therapeutic Potential by Combining Various Bioengineering Technologies

“…These 3D-bioprinted neural tissues expressed various neuronal markers (TUJ1, FOXA2, NURR1, TH, and PAX6) and glial markers (GFAP and O4) ( Sharma et al, 2020 ). Jury et al (2022) also developed 3D-bioprinted neural structures with hyaluronan and poly (ethylene glycol)-based hydrogels. The neuroepithelial stem cells within hydrogels undergo spontaneous differentiation into neural cells with significantly enhanced proliferation and viability.…”