Differentiation of Human Neural Progenitor Cells in Functionalized Hydrogel Matrices

Abstract

Hydrogel-based three-dimensional (3D) scaffolds are widely used in the field of regenerative medicine, translational medicine, and tissue engineering. Recently, we reported the effect of scaffold formation on the differentiation and survival of human neural progenitor cells (hNPCs) using PuraMatrix™ (RADA-16) scaffolds. Here, we were interested in the impact of PuraMatrix modified by the addition of short peptide sequences, based on a bone marrow homing factor and laminin. The culture and differentiation of the hNPCs in the modified matrices resulted in an approximately fivefold increase in neuronal cells. The examination of apoptotic and necrotic cells, as well as the level of the anti-apoptotic protein Bcl-2, indicates benefits for cells hosted in the modified formulations. In addition, we found a trend to lower proportions of apoptotic or necrotic neuronal cells in the modified matrices. Interestingly, the neural progenitor cell pool was increased in all the tested matrices in comparison to the standard 2D culture system, while no difference was found between the modified matrices. We conclude that a combination of elevated neuronal differentiation and a protective effect of the modified matrices underlies the increased proportion of neuronal cells.

Inhibition of BCL‐2 leads to increased apoptosis and delayed neuronal differentiation in human ReNcell VM cells in vitro

“…Anti-apoptotic BCL-2 increases and predominantly co-localizes with mitochondria during the differentiation of neural ReNcell VM progenitor cells, which is in agreement with published literature (Merry et al, 1994;Liedmann et al, 2012).…”

Alzheimer's in 3D culture: Challenges and perspectives

“…We have demonstrated that a powerful advantage of 3D culture for modeling AD is that it provides a local environment that promotes aggregation of β‐amyloid, which can trigger pathogenic cascades, including NFT . In addition, other studies have shown that 3D conditions more closely mimic in vivo environments and can accelerate neuronal differentiation and neural network formation . Neurons grown in a 2D environment are able to spread freely across the surface of the glass or plastic surface, but they have no support for vertical growth, which can lead to unintended apical‐basal polarity rather than the more accurate neuronal stellar morphology .…”

Modelling Alzheimer's disease: Insights fromin vivotoin vitrothree-dimensional culture platforms

“…The various advantages offered by 3D over 2D cellular cultures have been well acknowledged in recent literature (Pampaloni, Reynaud, & Stelzer, ; Ravi, Paramesh, Kaviya, Anuradha, & Solomon, ), especially in the provision of cues influencing cell structure, adhesion, proliferation, signalling, and mechanotransduction (Baker & Chen, ). This is also relevant in neuronal cultures, where a 3D substrate helps improve both cell yield (Liedmann, Frech, Morgan, Rolfs, & Frech, ) and cell differentiation (Zhang et al, ) in comparison with traditional 2D dishes. Figure highlights the key approaches utilized by researchers in realizing a 3D environment for in vitro cell culture, which is relevant in the context of neurodegenerative disease modelling and neurotherapeutics testing (Edelman & Keefer, ; Hopkins et al, ; Ko & Frampton, ).…”