Method Profile
A defined anesthetic exposure is followed by four weeks of culture before neuronal, glial, myelin and molecular endpoints are compared.
Investigation of persistent developmental responses to anesthetic exposure in human neural tissue.
Brain microphysiological spheroids derived from one human iPSC line, containing neural and glial lineages.
Nominal gas-phase 2.4% sevoflurane for four hours at week 8, followed by four additional culture weeks; carrier-gas control.
Human microtissues
iPSC-derived neural progenitors differentiated into multicellular bMPS cultures.
Measurements
Immunofluorescence, cell-marker ratios, myelin-associated process length, qPCR and mTOR-associated protein/phosphorylation measurements.
Interpret structural, expression and pathway-associated endpoints separately. Assay-specific cultures and differentiation batches remain distinct sampling units.
Compare delayed tissue phenotypes after sevoflurane versus carrier-gas exposure.
One iPSC line. Selected IF/WB cultures derive from two differentiation batches; vGlut1 qPCR uses six independent cultures.
Carrier-gas controls under otherwise matched culture conditions.
Exposed cultures showed lower myelin-associated measurements and presynaptic markers and increased apoptosis and mTOR-associated phosphorylation. Some protein-intensity changes were not significant.
A human brain microphysiological system showed lower myelin-associated measurements and presynaptic markers four weeks after sevoflurane exposure. The study combines neuronal, glial and pathway measurements within one iPSC background.
