Astrocytes Grown in Alvetex® Three Dimensional Scaffolds Retain a Non-reactive Phenotype

Abstract

Protocols which permit the extraction of primary astrocytes from either embryonic or postnatal mice are well established however astrocytes in culture are different to those in the mature CNS. Three dimensional (3D) cultures, using a variety of scaffolds may enable better phenotypic properties to be developed in culture. We present data from embryonic (E15) and postnatal (P4) murine primary cortical astrocytes grown on coated coverslips or a 3D polystyrene scaffold, Alvetex. Growth of both embryonic and postnatal primary astrocytes in the 3D scaffold changed astrocyte morphology to a mature, protoplasmic phenotype. Embryonic-derived astrocytes in 3D expressed markers of mature astrocytes, namely the glutamate transporter GLT-1 with low levels of the chondroitin sulphate proteoglycans, NG2 and SMC3. Embroynic astrocytes derived in 3D show lower levels of markers of reactive astrocytes, namely GFAP and mRNA levels of LCN2, PTX3, Serpina3n and Cx43. Postnatal-derived astrocytes show few protein changes between 2D and 3D conditions. Our data shows that Alvetex is a suitable scaffold for growth of astrocytes, and with appropriate choice of cells allows the maintenance of astrocytes with the properties of mature cells and a non-reactive phenotype.

Neuronal-glial populations form functional networks in a biocompatible 3D scaffold

“…Furthermore astrocyte morphology is notably different in such 3D cultures when compared with 2D monolayers. This is particularly interesting when the astrocyte morphology seen in 3D culture (extensive, intricate processes) was more consistent with astrocyte morphology in vivo than with that seen in comparator 2D monolayer cultures, as previously described [14]. However, given growing recognition of the 'quadpartite synapse' [12], investigations of microglia in 3D scaffolds such as Alvetex are warranted.…”

“…While a variety of commercial and bespoke scaffolds which support 3D cell culture are now in use [11] and neuronal-glial cultures have previously been grown on Alvetex membranes [14], our results demonstrate for the first time that spontaneously active, functional networks can be formed and monitored using murine central nervous system cells grown within this substrate. Specifically, we show viable murine mixed neuronal-glial cultures grow in 3D within the Alvetex substrates.…”

Sonic hedgehog signalling mediates astrocyte crosstalk with neurons to confer neuroprotection

“…For three‐dimensional (3D) transwell experiments, primary cortical astrocytes were seeded on Alvetex ® 6‐ and 12‐well hanging inserts (Reinnervate, Durham, UK) as previously described (Ugbode et al . ). The primary astrocyte cultures were of 95% purity.…”

Differentiation Patterns of Immortalized Human Meibomian Gland Epithelial Cells in Three-Dimensional Culture

“…As HMGEC cultured on ThinCert did not yield convincing results, we further tested Alvetex scaffolds allowing cells to proliferate three-dimensionally, thereby communicating more dynamically, 34 to differentiate in response to distinct stimuli 35,36 and to function in an appropriate microenvironment. 37,38 Histologic analysis demonstrated that HMGECs exhibited the same appearance regardless of their location within Alvetex.…”