Three-Dimensional Neural Spheroid Culture: AnIn VitroModel for Cortical Studies

Abstract

There is a high demand for in vitro models of the central nervous system (CNS) to study neurological disorders, injuries, toxicity, and drug efficacy. Three-dimensional (3D) in vitro models can bridge the gap between traditional two-dimensional culture and animal models because they present an in vivo-like microenvironment in a tailorable experimental platform. Within the expanding variety of sophisticated 3D cultures, scaffold-free, self-assembled spheroid culture avoids the introduction of foreign materials and preserves the native cell populations and extracellular matrix types. In this study, we generated 3D spheroids with primary postnatal rat cortical cells using an accessible, size-controlled, reproducible, and cost-effective method. Neurons and glia formed laminin-containing 3D networks within the spheroids. The neurons were electrically active and formed circuitry through both excitatory and inhibitory synapses. The mechanical properties of the spheroids were in the range of brain tissue. These in vivo-like features of 3D cortical spheroids provide the potential for relevant and translatable investigations of the CNS in vitro.

Developments in 3D neural cell culture models: the future of neurotherapeutics testing?

“…Self-assembled aggregate cultures, which encompass spheroids, embryoid bodies, neurospheres, and microtissues, can be formed through suspension culture, in concave microwells, on sponge-gel matrices, and by a variety of other methods [12][13][14][15][16][17][18]. Neural spheroids can be produced using primary neural cells or neural cell lines such as the SK-N-SH neuroblastoma cell line for brain tumor studies [12,13].…”

“…Neural spheroids can be produced using primary neural cells or neural cell lines such as the SK-N-SH neuroblastoma cell line for brain tumor studies [12,13]. Neural stem cell aggregates can also be used, but these aggregates have been criticized for being very sensitive to cell culture conditions, resulting in inconsistently differentiated cells even between experimental trials [14,15].…”

“…John.Frampton@dal.ca [12,14]. Scaffold-based cell cultures attempt to use synthetic and/ or natural biomaterials to reproduce neural tissue.…”

Spheroids Formation on Non‐Adhesive Surfaces by Liquid Overlay Technique: Considerations and Practical Approaches

“…[10] On the other side, Dingle and co-workers produced primary cortical spheroids that display several features of the in vivo cortical tissue (e.g., cell types, cell morphology, ECM production, and mechanical properties). [11] Moreover, spheroids have been also applied for the expansion and differentiation of pluripotent mesenchymal stem cells (MSCs), embryonic stem cells (ESC), and induced pluripotent stem cells (iPSC), as the cellular aggregates constituted by these cells (embryoid bodies) are able to mimic many cell differentiation features of early embryogenesis and play an important role in the in vitro differentiation process of cells. [12] Notwithstanding, spheroids are also a valuable 3D cellular model to replicate the features of human solid tumors, as discussed in different reviews.…”

Three-dimensional neuronal cell culture: in pursuit of novel treatments for neurodegenerative disease

“…[40] For example, Dingle et al generated neural spheroids from primary rat cortical tissues by means of micromolding, which were reproducible in size and cellular composition. [41] Spheroid characterization revealed resemblance to in vivo cortical tissue in terms of stiffness, neuronal electrophysiology, neural cell types, and morphology. Following on from this study, Boutin et al assessed the potential of this heterogeneous spheroid model to form capillary-like networks, the later playing an important role in protecting the neuronal environment.…”