Major changes of cell function and toxicant sensitivity in cultured cells undergoing mild, quasi-natural genetic drift

Gutbier, Simon; May, Patrick; Berthelot, Sylvie; Krishna, Abhimanyu; Trefzer, Timo B.; Behbehani, Mehri; Efremova, Liudmila V.; Delp, Johannes; Gstraunthaler, Gerhard; Waldmann, Tanja; Leist, Marcel

doi:10.1007/s00204-018-2326-5

Cited by 32 publications

(35 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, a 72‐h MPP+ exposure time did significantly reduce intracellular ATP, even at higher initial seeding densities (Figure S5a and S6). The elevated LC50 values for MPP+ we observed in our experiments appear to be consistent with a recent report comparing the sensitivity of LUHMES cells obtained from ATCC to LUHMES cells from the original provider . Genomic comparison identified that genetic drift may be occurring for this cell line, resulting in biochemical differences in differentiated cells that influence sensitivity to MPP+.…”

Section: Discussionsupporting

confidence: 91%

“…The elevated LC50 values for MPP+ we observed in our experiments appear to be consistent with a recent report comparing the sensitivity of LUHMES cells obtained from ATCC to LUHMES cells from the original provider. 56 Genomic comparison identified that genetic drift may be occurring for this cell line, resulting in biochemical differences in differentiated cells that influence sensitivity to MPP+. MPP+ cytotoxicity was associated with changes to the color of the culture medium from pink to yellow, indicating a lower pH.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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

Tam

Teixeira

et al. 2019

Biotechnology Progress

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

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

Tam

Teixeira

et al. 2019

Biotechnology Progress

View full text Add to dashboard Cite

show abstract

“…LUHMES cells have a normal human genome, a dopaminergic phenotype, neurotypical electrophysiological properties and sensitivity to PD-related neurotoxicants, including rotenone, 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) and its metabolite MPP + . Handling of LUHMES cell cultures was performed as previously described (Gutbier et al 2018a ; Scholz et al 2011 ). Briefly, the conditionally immortalized cells were maintained in proliferation medium (PM) that consisted of advanced Dulbecco's Modified Eagle Medium (DMEM)/F12, 2 mM l -glutamine, 1xN2 supplement (Invitrogen, CA, USA) and 40-ng/ml FGF-2, and were kept in a 5% CO 2 /95% air atmosphere at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

Design and evaluation of bi-functional iron chelators for protection of dopaminergic neurons from toxicants

et al. 2020

Self Cite

View full text Add to dashboard Cite

While the etiology of non-familial Parkinson's disease (PD) remains unclear, there is evidence that increased levels of tissue iron may be a contributing factor. Moreover, exposure to some environmental toxicants is considered an additional risk factor. Therefore, brain-targeted iron chelators are of interest as antidotes for poisoning with dopaminergic toxicants, and as potential treatment of PD. We, therefore, designed a series of small molecules with high affinity for ferric iron and containing structural elements to allow their transport to the brain via the neutral amino acid transporter, LAT1 (SLC7A5). Five candidate molecules were synthesized and initially characterized for protection from ferroptosis in human neurons. The promising hydroxypyridinone SK4 was characterized further. Selective iron chelation within the physiological range of pH values and uptake by LAT1 were confirmed. Concentrations of 10-20 µM blocked neurite loss and cell demise triggered by the parkinsonian neurotoxicants, methyl-phenyl-pyridinium (MPP +) and 6-hydroxydopamine (6-OHDA) in human dopaminergic neuronal cultures (LUHMES cells). Rescue was also observed when chelators were given after the toxicant. SK4 derivatives that either lacked LAT1 affinity or had reduced iron chelation potency showed altered activity in our assay panel, as expected. Thus, an iron chelator was developed that revealed neuroprotective properties, as assessed in several models. The data strongly support the role of iron in dopaminergic neurotoxicity and suggests further exploration of the proposed design strategy for improving brain iron chelation.

show abstract

“…Although variability of effects with different cell lines has been described (Colleoni et al 2014), no systematic analysis has been performed so far to evaluate their toxicological relevance. To eliminate the possibility of a differential genetic drift of cell lines in culture, which could interfere with toxicity assessment (Gutbier et al 2018), we used cell lines derived in one single laboratory, maintained in identical conditions, that have undergone a comparable moderate number of passages in culture (Sterthaus et al 2014). We nevertheless observed a significant level of variability (up to fivefold) in the responses of the four hESC lines to both compounds.…”

Section: Discussionmentioning

confidence: 99%

Accuracy, discriminative properties and reliability of a human ESC-based in vitro toxicity assay to distinguish teratogens responsible for neural tube defects

Feutz

Geyter

2019

Arch Toxicol

View full text Add to dashboard Cite

The poor correlation of developmental toxicity studies in animals with human outcome data has emphasized the need for complementary assays based on human cells and tissues. As neural tube defects represent an important proportion of congenital malformations, we evaluated here the accuracy of a human embryonic stem cell (hESC)-based assay to predict chemically induced disruption of neural tube formation. As teratogenic compounds, we used cyclopamine (CPA), valproic acid (VPA), ochratoxin A (OTA) and mycophenolic acid (MMF), all suspected or known inducers of human neural tube defects, as well as theophylline and saccharin as negative control compounds. We analyzed their effects on the ability of hES cells to give rise to neural precursors (expressing specific marker Nestin), to form neural tube-like structures (rosettes), and to express specific markers (Sox1, Otx2, Lix1, EvI1, Rspo3) during rosette formation. The results showed that various effects of the selected compounds on early neural development could be specifically revealed in vitro through related alterations of neurogenic differentiation of hESC. Furthermore, it was possible to discriminate toxicants acting at different time points during embryonic development and, therefore, responsible for distinct adverse effects on neural tube formation. By comparing four different hESC lines, we observed a significant (up to fivefold) variability of the line-dependent response to toxicants. We highlight at least two sources of variability: one related to the heterogeneity of hESC lines in culture (stemness/commitment profiles); the second to possible genetically determined differences in individual sensitivity to teratogens.Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

show abstract

Major changes of cell function and toxicant sensitivity in cultured cells undergoing mild, quasi-natural genetic drift

Cited by 32 publications

References 87 publications

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

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

Design and evaluation of bi-functional iron chelators for protection of dopaminergic neurons from toxicants

Accuracy, discriminative properties and reliability of a human ESC-based in vitro toxicity assay to distinguish teratogens responsible for neural tube defects

Contact Info

Product

Resources

About