SH‐SY5Y and LUHMES cells display differential sensitivity to MPP+, tunicamycin, and epoxomicin in 2D and 3D cell culture

Abstract

SH‐SY5Y and LUHMES cell lines are widely used as model systems for studying neurotoxicity. Most of the existing data regarding the sensitivity of these cell lines to neurotoxicants have been recorded from cells growing as two‐dimensional (2D) cultures on the surface of glass or plastic. With the emergence of 3D culture platforms designed to better represent native tissue, there is a growing need to compare the toxicology of neurons grown in 3D environments to those grown in 2D to better understand the impact that culture environment has on toxicant sensitivity. Here, a simple 3D culture method was used to assess the impact of growth environment on the sensitivity of SH‐SY5Y cells and LUHMES cells to MPP+, tunicamycin, and epoxomicin, three neurotoxicants that have been previously used to generate experimental models for studying Parkinson's disease pathogenesis. SH‐SY5Y cell viability following treatment with these three toxicants was significantly lower in 2D cultures as compared to 3D cultures. On the contrary, LUHMES cells did not show significant differences between growth conditions for any of the toxicants examined. However, LUHMES cells were more sensitive to MPP+, tunicamycin, and epoxomicin than SH‐SY5Y cells. Thus, both the choice of cell line and the choice of growth environment must be considered when interpreting in vitro neurotoxicity data.

Advanced 3D Models of Human Brain Tissue Using Neural Cell Lines: State-of-the-Art and Future Prospects

“…A number of studies compare cell responses to toxic compounds in 3D and 2D models developed with human neural cell lines. Differentiated neuroblastoma SH-SY5Y cells exhibit lower sensitivity to toxins when cultured in 3D constructs than in 2D ones [ 66 , 67 ]. Furthermore, 3D models allow the inspection of the influence of the micro-environment on the cell response and sensitivity to neurotoxins [ 84 ].…”

LUHMES Cells: Phenotype Refinement and Development of an MPP+-Based Test System for Screening Antiparkinsonian Drugs

“…In a number of studies devoted to the development of a PD model on LCs using MPP + , it is emphasized that the ATCC cells at our disposal are much less sensitive to the toxic effect of MPP + than the UKN cells [10,32,63,64]. This is mainly due to a much lower level (about 50 times) of DAT expression in ATCC cells compared to UKN cells [32].…”

Biomaterials-based strategies for in vitro neural models

“…276 These models generate more physiologically accurate responses to applied pharmaceuticals. 16 Further strategies that incorporate chemical cues of the native tissues or co-cultures with in vivo support cells such as astrocytes, oligodendrocytes or Schwann cells drive the models even closer to the complex biochemical environment of neural tissues in situ. 153 In this context, organoids and spheroids utilize intrinsic cell-cell interactions of pluripotent stem cells cultured as embryoid bodies 277 or within a 3D matrix that is chemically close to native ECM.…”

“…Taken together, when neural structures or pathologies are studied with in vitro systems which present the aforementioned limitations, deviations from the native cell behavior (e.g. flattened growth cones, 15 restricted neurite outgrowth and altered response to administered drugs 16 ) are observed. In addition, conventional models further restrict the investigation of neurodegenerative pathologies since cell-secreted neurotoxic factors are diluted in the culture medium and eventually removed during media changes, as opposed to creating highly concentrated nucleation sites in the extracellular space during in vivo progression and propagation of the pathologies.…”