Neuro-differentiated Ntera2 cancer stem cells encapsulated in alginate beads: First evidence of biological functionality

- Embryonic cancer stem NTera2 cells (NT2) were successfully encapsulated within alginate beads.
- The used procedure for NT2 encapsulation allowed to preserve the cell viability over time.
- The functionality of the encapsulated NT2 cells was confirmed by their dopamine production capability.
- Higher starting cell concentrations favoured the NT2 cell aggregation and organization in clusters.
- The alginate capsules/NT2 systems may be potentially employable as an alternative cell therapy for the Parkinson disease.

The present communication investigates an application of alginate encapsulation technology to the differentiation of the embryonic cancer stem NTera2 cells (NT2) into dopamine-producing cells. The encapsulation of cells in polymeric beads allows their immune isolation and makes them eligible for transplantation, thus representing a promising biotech tool for the delivery of biologically active compounds to the brain. The polysaccharide alginate is one of the most commonly used material for this procedure since it is well tolerated by various tissues, including the brain. Two different initial cell concentrations (i.e. 0.5 ∗ 106/ml and 1.0 ∗ 106/ml) were tested, in order to identify which one could better reflect the homogeneous cell distribution into the alginate beads and guarantee a good cell viability at different times of culture. As evidenced, the higher number of cells promoted the formation of clusters resulting in a better interaction among encapsulated cells and the subsequent promotion of mitotic activity. The distribution of alive/dead cells into the alginate beads was verified and followed at different time points through the fluorescein diacetate/propidium iodide (FDA/PI) staining, confirming the presence of living neuronal positive cells, as determined from fluorescence microscopy imaging. The functionality of the encapsulated NT2 cells was confirmed by their dopamine production capability as assessed by UV-Vis spectrophotometric analysis and by liquid chromatography-mass spectrometry (LC-MS). The NT2/microspheres system can be considered a groundbreaking experimental procedure, a functionally active platform, able to produce and release dopamine, and thus potentially exploitable for therapy in Parkinson's disease.

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