Establishment of human cerebral organoid systems to model early neural development and assess the central neurotoxicity of environmental toxins.

JOURNAL/nrgr/04.03/01300535-202501000-00032/figure1/v/2024-05-14T021156Z/r/image-tiff Human brain development is a complex process, and animal models often have significant limitations. To address this, researchers have developed pluripotent stem cell-derived three-dimensional structures, known as brain-like organoids, to more accurately model early human brain development and disease. To enable more consistent and intuitive reproduction of early brain development, in this study, we incorporated forebrain organoid culture technology into the traditional unguided method of brain organoid culture. This involved embedding organoids in matrigel for only 7 days during the rapid expansion phase of the neural epithelium and then removing them from the matrigel for further cultivation, resulting in a new type of human brain organoid system. This cerebral organoid system replicated the temporospatial characteristics of early human brain development, including neuroepithelium derivation, neural progenitor cell production and maintenance, neuron differentiation and migration, and cortical layer patterning and formation, providing more consistent and reproducible organoids for developmental modeling and toxicology testing. As a proof of concept, we applied the heavy metal cadmium to this newly improved organoid system to test whether it could be used to evaluate the neurotoxicity of environmental toxins. Brain organoids exposed to cadmium for 7 or 14 days manifested severe damage and abnormalities in their neurodevelopmental patterns, including bursts of cortical cell death and premature differentiation. Cadmium exposure caused progressive depletion of neural progenitor cells and loss of organoid integrity, accompanied by compensatory cell proliferation at ectopic locations. The convenience, flexibility, and controllability of this newly developed organoid platform make it a powerful and affordable alternative to animal models for use in neurodevelopmental, neurological, and neurotoxicological studies.

Modeling early human cortical development and evaluating neurotoxicity with a forebrain organoid system.

The complexity and subtlety of brain development renders it challenging to examine effects of environmental toxicants on human fetal brain development. Advances in pluripotent cell-derived organoid systems open up novel avenues for human development, disease and toxicity modeling. Here, we have established a forebrain organoid system and recapitulated early human cortical development spatiotemporally including neuroepithelium induction, apical-basal axis formation, neural progenitor proliferation and maintenance, neuronal differentiation and layer/region patterning. To explore whether this forebrain organoid system is suitable for neurotoxicity modeling, we subjected the organoids to bisphenol A (BPA), a common environmental toxicant of global presence and high epidemic significance. BPA exposure caused substantial abnormalities in key cortical developmental events, inhibited progenitor cell proliferation and promoted precocious neuronal differentiation, leading premature progenitor cell depletion and aberrant cortical layer patterning and structural organization. Consistent with an antagonistic mechanism between thyroid hormone and BPA, T3 supplementation attenuated BPA-mediated cortical developmental abnormalities. Altogether, our in vitro recapitulation of cortical development with forebrain organoids provides a paradigm for efficient neural development and toxicity modeling and related remedy testing/screening.

Variable metal resistance of P. putida CZ1 biofilms in different environments suggests its remediation application scope.

Pseudomonas putida is potentially used in the bioremediation of heavy metals (HMs). Its response to different HMs in different environments is still not fully understood. This study investigated resistance against 12 kinds of metals by P. putida CZ1 planktonic cells and its biofilm in LB and mineral medium (MM). P. putida CZ1 biofilms have high resistance and accumulation capacity for Cu2+, Zn2+, Pb2+, Fe3+, Mn2+, Al3+ and Ni2+, but less resistance to Co2+, Cd2+, Cr2O72-, Ag+ and Hg2+. Biofilms were 2-8 times more resistant to Cu2+ and Zn2+ than planktonic cells. There was a strong correlation between the P content and the accumulation of Cu2+, Zn2+, Fe3+, Mn2+, Pb2+, Ni2+and Al3+ respectively. Confocal laser scanning microscopy (CLSM) combined with live/dead staining study found that cells in the biofilms can keep viable after 36 h under MIC of Cu2+ or Zn2+ both in LB and MM. When the metal concentration increased, cells can be killed gradually. For Cu2+, Zn2+, Fe3+, Mn2+, Pb2+ and Ni2+, higher resistance was found in MM (2-4 times higher) than in LB and higher accumulation of these metals were also found in MM. P. putida CZ1 biofilm cultured in MM with citric acid as carbon source had stronger resistance and accumulation ability to Cu2+, Zn2+, Pb2+, Fe3+, Mn2+, and Ni2+. This suggested that P. putida CZ1 had greater remediation potential for these metals in organic acid rich environments.