An In Vitro Partial Lesion Model of Differentiated Human Mesencephalic Neurons: Effect of Pericyte Secretome on Phenotypic Markers

Abstract

Parkinson’s disease (PD) is characterised by the progressive degeneration of dopaminergic (DA) neurons in the substantia nigra pars compacta. Post-mortem data suggests that the loss of DA markers may long precede the cell death, leaving a window to rescue the DA phenotype. Screening for potential neuroprotective or restorative therapies, however, requires that partial lesions of DA neurons can be modelled in vitro. In order to establish a partial lesion model of DA neurons in vitro, we evaluated the effects of different exposure times to 1-methyl-4-phenylpyridinium (MPP+) and 6-hydroxydopamine (6-OHDA) on the cell survival and DA marker expression using DA neurons derived from the Lund human mesencephalic (LUHMES) cell line. We show that 24-h incubation with 50 μM of MPP+ or 6-h incubation with 100 μM of 6-OHDA leads to a significant decrease in the protein expression of DA markers without affecting overall cell death, consistent with a mild DA lesion. Using conditioned medium of human brain–derived pericytes stimulated with platelet-derived growth factor BB (PDGF-BB), we demonstrate a significant upregulation of DA markers. In conclusion, we provide an experimental model of an in vitro DA neuron partial lesion suitable to study different molecules and their potential neuroprotective or neurorestorative effects on the DA phenotype. We provide evidence that the secretome of brain pericytes stimulated via PDGF-BB/PDGFRβ affects DA marker expression and may represent one possible mechanism contributing to the neurorestoration previously observed in PD by this growth factor.

A New Method for the Visualization of Living Dopaminergic Neurons and Prospects for Using It to Develop Targeted Drug Delivery to These Cells

“…This conclusion was strongly supported by the fact that we did not observe any changes in LUHMES cells with a MTT test even after 24 h of incubation with GBR-BP. Under the same incubation but with MPP+ (concentration 5 μM or 10 μM, 12-h incubation) instead of GBR-BP, a massive degeneration was observed in LUHMES cells [ 49 , 50 ].…”

Probabilistic cell seeding and non-autofluorescent 3D-printed structures as scalable approach for multi-level co-culture modeling

“…All three proteins show increased expression in differentiated LUHMES cells and are used as maturity markers. [ 33 , 39 , 40 ] The quantitative analysis showed that TH is significantly downregulated in all co-culture conditions (LUHMES cells growing on the bottom substrate layer and inside pillar cavities) compared to the monoculture ( Fig. 6 a).…”

Insights Into the Role of Platelet-Derived Growth Factors: Implications for Parkinson’s Disease Pathogenesis and Treatment

“…This demonstrates that brain pericytes may play a role in the pathogenesis of PD and may be a target for PDGF-BB treatment of PD neural recovery mechanisms (Padel et al, 2016). Subsequently, using an in vitro model of dopaminergic injury, Gaceb et al (2020) demonstrated that PDGF-BB/PDGFRβ-mediated brain pericyte secretion affects the expression of dopamine markers, which may shed light on the mechanism by which PDGF-BB promotes neural recovery in PD (Gaceb et al, 2020). These studies provide inspiration for future related research in this field.…”