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In the in vitro model here proposed we used the bEnd3 cell line as brain endothelial cells to mimic the blood brain barrier (BBB) cultured on dextran microbeads ranging from 67 μm to 80 μm in size (Cytodex) and the primary human umbilical vein cells (HUVEC) for comparison.
The effect of cruzipain on viral replication in macrophages is clear but all the analyses have been conducted on an in vitro model here.
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Thus, we developed astrocyte-like and neuron-like in vitro models here, as 3D monotypic (GL15 cells only; SH-SY5Y cells only) and heterotypic (cocultures of both GL15 and SH-SY5Y cells) cell cultures in the RCCS bioreactor.
This is also likely to be the case in the in vitro model used here.
Since the in vivo model is much closer to reality, even taking the limitations of a mice model into account, we suspect that the in vitro decrease in infection towards 96 hours post infection was a consequence of the limitations of the in vitro model used here.
Whether this finding reflects different selection processes in the field in Cambodia and during the in vitro model used here is unclear.
While detailed in vivo studies of the host response are now required, the in vitro model described here will allow responses to specific modulators (such as therapeutics) to be investigated.
Implications and future directions The iPSC-based in vitro model described here allows the evaluation of the effects of patient-specific ALS-associated FUS mutations on ALS-relevant cell types, e.g. motoneurons.
The in vitro model presented here could also facilitate the search for strategies to prevent and/or reduce the tissue and organ damage caused by hypoxia and ischemia-reperfusion injury.
The in vitro model used here showed that when neurons and d-hDPSCs were cultured together within a 3D hydrogel environment, the d-hDPSCs spontaneously ensheathed neurites and, in some cases, generated myelin structures, both characteristics that would be associated with neuron-Schwann cell interactions in vivo.
As a future perspective, the iPSC-based in vitro model described here, derived from patients by reprogramming or generated by site-directed mutagenesis, might greatly improve our understanding of the molecular basis of MN death and possibly help finding new therapeutic targets for ALS.
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