Soft surfaces promote astrocytic differentiation of mouse embryonic neural stem cells via dephosphorylation of MRLC in the absence of serum

Abstract

Astrocytes, which can be obtained from neural stem cells (NSCs) by adding serum and/or recombinant proteins in culture media or by passaging NSCs repeatedly, are expected to be applicable in regenerative medicine for the treatment of neurodegenerative diseases. However, astrocytes obtained using existing methods are costly and have poor quality. The stiffness of culture surfaces has been reported to affect astrocytic differentiation of adult NSCs. However, the influence of surface stiffness on astrocytic differentiation of embryonic NSCs has not yet been reported. In this study, we showed that astrocytic differentiation of embryonic NSCs was increased on soft surfaces (1 kPa and 12 kPa) compared with the NSCs on stiff surfaces (2.8 GPa) in serum-free condition. Furthermore, di-phosphorylated myosin regulatory light chain (PP-MRLC) was decreased in embryonic NSCs cultured on the soft surfaces than the cells on the stiff surfaces. Additionally, astrocytic differentiation of embryonic NSCs was induced by a Ras homolog associated kinase (ROCK) inhibitor, which decreased PP-MRLC in NSCs. These results suggest that decreasing the PP-MRLC of embryonic NSCs on soft surfaces or treating NSCs with a ROCK inhibitor is a good method to prepare astrocytes for application in regenerative medicine.

Hydrogel platform facilitating astrocytic differentiation through cell mechanosensing and YAP-mediated transcription

“…2 B–C), which was therefore used in all the subsequent experiments to represent the hydrogel. Remarkably, comparing to the PS substrate (elastic modulus in the order of GPa) [ 8 , 24 ], astrocytic differentiation on the alginate hydrogel was drastically enhanced, as revealed by immunofluorescence of S100B and GFAP ( Fig. 2 E–F, S2), and verified by immunoblots ( Fig.…”

“…Mechanical signals, such as matrix elasticity, regulate NSC differentiation into astrocytes. While compliant substrates with elastic modulus <1 kPa preferably direct NSCs into neuronal cells, relatively stiffer substrates (1–10 kPa) preferably lead to astrocytes and oligodendrocytes [ 7 , 8 ]. A study using human NSCs (hNSCs) has demonstrated that the stretch-activated Ca 2+ channel, Piezo1, promotes neurogenesis and suppresses astrogenesis, possibly via Yes-associated protein (YAP) [ 9 ].…”

Canonical and noncanonical Wnt signaling: Multilayered mediators, signaling mechanisms and major signaling crosstalk

“… 46 , 47 ROCK activates the myosin II regulatory light chain (MRLC). 48 , 49 CapZIP, MRLC, DIA1, and profilin all contribute to actin polymerization, which is vital to cell polarity and migration. 39 The Celsr1 receptor appears to function similarly to the Fz receptor, where Wnt binding ultimately causes activation of Dvl to stimulate the same signaling cascade.…”

Sequencing-based study of neural induction of human dental pulp stem cell

“…This property facilitates NSC cultivation in vitro, making them extremely attractive for investigating neurogenesis and the progenitor-mediated development of various cell lineages. Moreover, NSC transplantation is presumed to exert therapeutic effects in neurodegenerative disorders, cerebrovascular diseases, and traumatic brain or spinal cord injuries via regeneration, repair, or enhancement of central nervous system functions [2][3][4][5]. Furthermore, NSCs substantially contribute to the establishment of disease models and drug and toxicity screening research [6].…”