Smart Neurobiology with BIOFLOAT
The brain is composed of a complex 3D interconnected system of different cells and extracellular matrixes (ECM). To mimic its 3D organization in-vivo, in-vitro culturing presents a unique challenge. The culture conditions should mimic the appropriate microenvironment with topographical cues that influences proliferation, differentiation and migration of cells, resulting in different protein and gene expression profiles (1-4). 3D cultures achieve such conditions, providing an in-vivo like environment, on a well-defined platform.
Scaffold free Astrocyte spheroids
To this end, 3D culturing systems have come along way at improving 3D cell culture methods that mimic the native state conditions, and provide reliable, reproducible cultures that can be used in drug discovery studies, as well as track disease progression. Spheroid cultures based on a scaffold free system, prevents the cell from exposure to foreign materials, and enables formation of native cells populations with its own extracellular matrixes. For example, SFFV2 (immortalized astrocytes) have been successfully used in spheroid culture to study the role of astrocytes in health and disease (5). Astrocytes are the most numerous cell populations in the brain and are found in direct vicinity of neuronal synapsis; they are necessary for neurotransmitter recycling but also control synapse formation, function, and removal. To this end, astrocytes 3D spheroid cultures can be generated using BIOFLOAT 96-well plates. These plates are produced with a special polymeric coating rendering the cell culture surface cell repellent and fully inert.
Human immortalized astrocyte spheroids (SFFV2) with BIOFLOAT 96 well plates
When comparing with other benchmark products, Human immortalized astrocytes (SFFV2) cells cultured on BIOFLOAT 96 well plates indicated cell aggregation within the first 1 hour-24 hours post seeding. Spheroid generation time is crucial for time-dependent experiments as well as for drug testing studies in which reducing days of analysis can shorten the preclinical phase. The astrocytes cultivated on BIOFLOAT coated plates, were of uniform shape and consistency, another essential feature necessary to reduce data variability. BIOFLOAT plates also outperformed both Benchmark products tested in the viability of generated spheroids, which qualifies them for applications in which the performance is evaluated by reduced viability such as drug testing systems (5).
Establishment of a well defined, reliable and modifiable 3D astrocyte cultures has a wide range of applications and is essential to answer fundamental questions in the field of neurodegenerative diseases or as model systems for toxicological screening purposes. BIOFLOAT coated 96 well plates provides an easy to use platform with fast growth of spheroids with uniform consistency and high viability. This manner of 3D cell culture
platform can provide the ability to overcome the gap between current 2D based model systems with insufficient reproducibility or transferability to humans as well as the lack of human donor cells.
References
1) Balgude, A.P., Yu, X., Szymanski, A., and Bellamkonda, R.V. Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures. Biomaterials 22, 1077, 2001.
2) Li, G.N., Livi, L.L., Gourd, C.M., Deweerd, E.S., and Hoffman-Kim, D. Genomic and morphological changes of neuroblastoma cells in response to three-dimensional matrices. Tissue Eng 13, 1035, 2007.
3) Bruder, J.M., Lee, A.P., and Hoffman-Kim, D. Biomimetic materials replicating Schwann cell topography enhance neuronal adhesion and neurite alignment in vitro. J Biomater Sci Polym Ed 18, 967, 2007.
4) Baker, B.M., and Chen, C.S. Deconstructing the third dimension—how 3D culture microenvironments alter cellular cues. J Cell Sci 125, 3015, 2012.
5) https://facellitate.com/wp-content/uploads/White-Paper-Astrocytes_final.pdf
