Comparative Human and Rat “Neurosphere Assay” for Developmental Neurotoxicity Testing

Baumann, Jenny; Barenys, Marta; Gaßmann, Kathrin; Fritsche, Ellen

doi:10.1002/0471140856.tx1221s59

Cited by 39 publications

(51 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rat neural progenitor cells [rNPCs, postnatal day (PND) 5] were prepared time-matched to hNPCs (Clancy et al 2007) as described previously (Baumann et al 2014).…”

Section: Cell Culturementioning

confidence: 99%

“…Differentiation was initiated by growth factor withdrawal in differentiation medium and plating onto poly-d-lysine (PDL)/laminin-coated chamber slides as described previously (Baumann et al 2014). For details, see Supplementary Material.…”

Section: Cell Culturementioning

confidence: 99%

“…In every experiment, mitochondrial reductase activity was assessed in the same wells than the specific endpoint evaluations as previously described (Baumann et al 2014). For details, see Supplementary Material.…”

Section: Cell Viability Assaymentioning

confidence: 99%

“…In this respect, we previously developed in vitro models for DNT key event screening, which are based on primary human and rat neural progenitor cells grown as neurospheres (Baumann et al 2014). They are able to mimic basic processes of early fetal brain development such as proliferation, migration and differentiation to neural effector cells (Fig.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Comparative human and rat neurospheres reveal species differences in chemical effects on neurodevelopmental key events

et al. 2015

Self Cite

View full text Add to dashboard Cite

The developing brain is highly vulnerable to the adverse effects of chemicals, resulting in neurodevelopmental disorders in humans. Currently, animal experiments in the rat are the gold standard for developmental neurotoxicity (DNT) testing; however, these guideline studies are insufficient in terms of animal use, time and costs and bear the issue of species extrapolation. Therefore, the necessity for alternative methods that predict DNT of chemicals faster, cheaper and with a high predictivity for humans is internationally agreed on. In this respect, we developed an in vitro model for DNT key event screening, which is based on primary human and rat neural progenitor cells grown as neurospheres. They are able to mimic basic processes of early fetal brain development and enable an investigation of species differences between humans and rodents in corresponding cellular models. The goal of this study was to investigate to what extent human and rat neurospheres were able to correctly predict the DNT potential of a well-characterized training set of nine chemicals by investigating effects on progenitor cell proliferation, migration and neuronal differentiation in parallel to cell viability, and to compare these chemical responses between human and rat neurospheres. We demonstrate that (1) by correlating these human and rat in vitro results to existing in vivo data, human and rat neurospheres classified most compounds correctly and thus may serve as a valuable component of a modular DNT testing strategy and (2) human and rat neurospheres differed in their sensitivity to most chemicals, reflecting toxicodynamic species differences of chemicals.

show abstract

“…Rat neural progenitor cells [rNPCs, postnatal day (PND) 5] were prepared time-matched to hNPCs (Clancy et al 2007) as described previously (Baumann et al 2014).…”

Section: Cell Culturementioning

confidence: 99%

Section: Cell Culturementioning

confidence: 99%

Section: Cell Viability Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Comparative human and rat neurospheres reveal species differences in chemical effects on neurodevelopmental key events

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…for rats, where there is sufficient data on mode of action of compounds in vivo, then the equivalent cell system will with a high probability be also predictive for different species including humans. This approach has been tested with primary NPs as well as primary rat cortical and hESC-derived neurons for different DNT-specific endpoints (Baumann et al, 2014, Gassmann et al, 2010, Harrill et al, 2011. The endpoint group 'NS/PC Endpoints' is mainly comprised of different methods assessing stemness-or progenitor cell markers or morphological aspects like neural rosette formation.…”

Section: Neural Proliferationmentioning

confidence: 99%

Literature review on in vitro and alternative Developmental Neurotoxicity (DNT) testing methods

Fritsche

Alm

Baumann

et al. 2015

EFSA Supporting Publications

Self Cite

View full text Add to dashboard Cite

The goal of this systematic review performed under a contract with EFSA was the evaluation of information on assessment methods in the field of developmental neurotoxicity (DNT). Therefore, a systematic and comprehensive literature search and collection of scientific literature and all other relevant grey literature and website information (in English) from past 20 years until mid of April 2014 on the state of the art of in vivo DNT testing methods including novel and alternative non-mammalian models, in vitro test methods, in silico methods, read across and combination of testing methods in test batteries was performed. This systematic review identified a variety of methods covering early and later stages of neurodevelopment that have the ability to predict DNT of chemicals. Few in vivo models alternative to OECD TG 426 were identified. In general, the available published in vivo data is scattered and heterogeneous, making comparisons across exposure paradigms and compounds very difficult. Thus, to make more useful evaluations, publications with more consistent experimental designs are needed, and more focused in vivo-in vitro comparisons are needed to establish useful effect biomarkers and testing models. From the alternative methods, a testing strategy covering early and later neurodevelopmental stages (from stem cell to zebrafish larvae motor behaviour) can be assembled. For gaining regulatory acceptance, definition of biological application domains of alternative methods by performing e.g. in vitro -in vivo validation is needed. Moreover, protocols for cell-based and zebrafish assays need international standardization. With such standardized protocols, the test battery needs to be tested for its sensitivity and specificity by testing concentration-responses of known DNT positive and negative compounds across the different assays. Such data might then be used either for regulatory decision-making or compound prioritization in favour of a reduction of rodent guideline in vivo testing.

show abstract

Rabbit neurospheres as a novel in vitro tool for studying neurodevelopmental effects induced by intrauterine growth restriction

Barenys

Illa

Hofrichter

et al. 2020

Stem Cells Translational Medicine

Self Cite

View full text Add to dashboard Cite

The aim of this study was to develop a rabbit neurosphere culture to characterize differences in basic processes of neurogenesis induced by intrauterine growth restriction (IUGR). A novel in vitro neurosphere culture has been established using fresh or frozen neural progenitor cells from newborn (PND0) rabbit brains. After surgical IUGR induction in pregnant rabbits and cesarean section 5 days later, neural progenitor cells from both control and IUGR groups were isolated and directly cultured or frozen at −80 C. These neural progenitor cells spontaneously formed neurospheres after 7 days in culture. The ability of control and IUGR neurospheres to migrate, proliferate, differentiate to neurons, astrocytes, or oligodendrocytes was compared and the possibility to modulate their responses was tested by exposure to several positive and negative controls. Neurospheres obtained from IUGR brains have a significant impairment in oligodendrocyte differentiation, whereas no significant differences are observed in other basic processes of neurogenesis. This impairment can be reverted by in vitro exposure of IUGR neurospheres to thyroid hormone, which is known to play an essential role in white matter maturation in vivo. Our new rabbit neurosphere model and the results of this study open the possibility to test several substances in vitro as neuroprotective candidates against IUGR induced neurodevelopmental damage while decreasing the number of animals and resources and allowing a more mechanistic approach at a cellular functional level.

show abstract

Comparative Human and Rat “Neurosphere Assay” for Developmental Neurotoxicity Testing

Cited by 39 publications

References 31 publications

Comparative human and rat neurospheres reveal species differences in chemical effects on neurodevelopmental key events

Comparative human and rat neurospheres reveal species differences in chemical effects on neurodevelopmental key events

Literature review on in vitro and alternative Developmental Neurotoxicity (DNT) testing methods

Rabbit neurospheres as a novel in vitro tool for studying neurodevelopmental effects induced by intrauterine growth restriction

Contact Info

Product

Resources

About