for the assessment of developmental neurotoxicity

Overview

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Key numbers

The worker extracted the full PDF text with layout preservation twice and compared extraction hashes before commit. The following lines are copied from numeric/table-bearing regions of the PDF and retain the source units and wording where legible:

• iPSCs were passaged every 4 - 5 days when it reached 80 - 90% confluency using the StemPro®
• GlutaMAX (35050-038, Invitrogen), 1% (v/v) MEM-nonessential amino acids (MEM-NEAA;
• M7145, MilliporeSigma), 3% hESC quality FBS (10439001, ThermoFisher), 100 µM 2-
• medium (NIM) consisting of DMEM-F12, 1% (v/v) N2 supplement (17502001, ThermoFisher), 1%
• equal volume of DMEM-F12 and neurobasal medium (21103049, Fisher Scientific) with 0.5% (v/v)
• consists of an equal volume of DMEM-F12 and neurobasal medium with 0.5% (v/v) N2
• NEAA, 1% (v/v) penicillin-streptomycin, 100 µM 2-mercaptoethanol and 2% (v/v) of Matrigel.
• serially diluted in the NIM to prepare six different dosages. The concentration range used was 4
• from 0% to 100% plotted against the logarithm of test concentrations, we normalized the
• luminescence intensities of all cerebral organoid spots with the luminescence intensity of a 100%
• The cerebral organoids were fixed with 4% paraformaldehyde (PFA) overnight at 4°C, washed
• with PBS for 10 minutes each for three times, and incubated with 30% sucrose overnight at 4°C.
• laminar flow for 30 minutes before staining. The slides were fixed in 10% buffered formalin for 20
• organoids on the pillar plate were rinsed with 1x PBS and treated with 4% PFA in the deep well
• plate overnight at 4°C. The organoids were permeabilized with 1% Triton X-100 in PBS for 1 hour
• at room temperature and then exposed to a blocking buffer (consisting of 4% normal donkey
• serum in 1x PBS with 0.5% Triton X-100) overnight at 4°C. Primary antibodies were diluted to
• 0.2% Triton X in PBS three times for 20 minutes each and incubated with appropriate secondary
• with 0.5% Triton X-100 in PBS three times for 20 minutes each and incubated with 1 µg/mL DAPI
• in 0.5% Triton X-100 in PBS for 30 minutes. This was followed by final washing with PBS three
• 4 AM cell permeant dye (F14201, ThermoFisher) for calcium imaging. Briefly, the culture medium
• phosphate dehydrogenase (GAPDH). The primers used are listed in Supplementary Table 3.
• suspension was collected in a 15 mL tube and fixed with 4% paraformaldehyde (PFA) for 10
• minutes at room temperature. The cells were permeabilized with 0.1% Triton X-100 in PBS for 15
• minutes, followed by blocking with 5% normal donkey serum (NDS) in 0.1% Triton X-100 in PBS
• generate cerebral organoids reproducibly with the coefficient of variation (CV) in the range of 7 -
• 19% in three trials 36. To demonstrate the cerebral organoid culture uniformly across the entire
• organoids on day 31 was in the range of 1000 - 1400 µm in diameter, which was significantly
• bigger than the size of statically cultured cerebral organoids in the range of 700 - 900 µm with the
• CV value less than 15% (Figs. 2C & 2D). The size of the organoids after 63 days of static and
• 31 cerebral organoids cultured in static (SC) and dynamic (DC) conditions. n = 36 per condition.
• kit. n = 18. (F) Neural biomarker expression in dynamically cultured cerebral organoids at days 0,
• 0.001, ** for p < 0.01, * for p < 0.05, n. s. = not significant (p > 0.05). n = 10 - 12 per qPCR run.
• assess the percentage of the cells expressing PAX6, FOXG1, TBR2, and CTIP2. n = 8.
• < 0.001, ** for p < 0.01, * for p < 0.05, n. s. = not significant (p > 0.05). n = 10 - 12 per qPCR run.
• as CTIP2 and SOX2 staining revealed the distinctive arrangement of cortical neurons in the
• 3 acid was noncytotoxic at the concentrations tested. n = 6 per dosage.
• compound-treated organoids (control) for apoptotic biomarkers. n = 10 - 12 per qPCR run. BCL2
• exposed to 250 nM methylmercury. n = 12.

Methods (brief)

• collected cells were further dissociated into single cells through pipetting and seeded in an
• suspension was collected in a 15 mL tube and fixed with 4% paraformaldehyde (PFA) for 10
• for p < 0.01, * for p < 0.05, and ns = not significant (p > 0.05). Sample sizes are indicated in the
• 24 H. Kim, R. Xu, R. Padmashri, A. Dunaevsky, Y. Liu, C. F. Dreyfus and P. Jiang, Stem Cell

Implications

This page makes the source discoverable for category-level evidence routing. Values remain source-native and should be used only with the stated matrix, species, basis, geography, and censoring context from the paper. The page does not convert total mercury to methylmercury or use total arsenic as inorganic arsenic.

Wiki pages this source may touch

• Fish — marine, non-predatory (sardines, anchovies, salmon, cod)
• Mercury

Verification notes

• Identity check: DOI, raw handle, candidate cite-key, and SHA-256 were compared against existing wiki/sources/ pages before creation.
• Full-PDF read: pdftotext -layout was run on the full PDF twice; extracted text hashes matched before the page was written.
• Numeric verification: numeric/table-bearing lines were selected mechanically from the verified extraction and preserved without unit conversion or rounding.
• Brand firewall: the worker skips PDFs when extracted numeric lines appear brand/manufacturer-sensitive; this page contains category-level or species-level evidence only.
• HMTc firewall: no threshold, percentile, pass/fail, clean/dirty, or certification math is stated.

Update history

The five most recent substantive edits to this page, classified major (evidence or structure moved), correction (a published value or statement was wrong and has been fixed), or minor (narrative rewritten without changing the underlying evidence). Each description is derived from what the edit did to this page; the linked commit is the authoritative record, routine regeneration passes are excluded, and the full version history lives in git. When DOI minting comes online (see schema docs), each entry below will also link to a version-pinned DataCite DOI.