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Grace, A. A. & Bunney, B. S. Intracellular and extracellular electrophysiology of nigral dopaminergic neurons–1. Identification and characterization. Neuroscience 10, 301–315 (1983).Higher numbers of mDA neurons are generated in 3D, with marker expression profiles indicative of a midbrain fate

With soluble media conditions 1, 4 adapted to 3D, we engineered a thermoresponsive, synthetic hydrogel system for scalable induction and differentiation of mDA neurons for biomedical applications such as Parkinson’s disease therapy. After 25 days of differentiation, a timepoint previously found to be optimal for cell transplantation in PD models 1, we report the generation of a ~2-fold higher proportion of mDA neurons ( Fig. 2b), and ~5 fold higher numbers of cells generated per volume of medium consumed ( Fig. 2a), in the 3D platform compared to a 2D control. A crucial benchmark for clinically relevant mDA neurons is TH expression. The majority in vitro differentiation studies report 15–30% TH positive cells after 25–45 days of differentiation on 2D platforms 1, 6, 40, 41, and in 2D we similarly find 20% TH+ cells at D25. In contrast, in the 3D hydrogel we generated a higher quality, purer mDA neuronal population, with almost double the percentage of TH+ cells (37%) compared to our 2D control. A more enriched population of mDA neurons entails a more efficient use of resources, may increase the therapeutic chances of success, and reduces the risk of side effects from contaminating cell types 42. Finally, medium (including small molecules and growth factors) is one of the most resource intensive components in cell production, and notably 100,000 neurons were generated per ml of medium used in 3D, compared to 40,000/ml on 2D.

Delivery & returns

It's built in slices which are connected to the one below by four studs around the central pillar. This enables them to be attached at angles other than 90 and 45 degrees, which helps give the finished conifer an organic appearance.