Evidence map›Paper›PMID 40466868›Full record

ArticleJournal of neuroscience methods2025

Under-oil neuronal cell culture: Enhanced system stability, yield, and modulated oxygen environment.

Nai-Wen Liang, Ligi Milesh, Karina N Buttram, Jessica L Park, Jing Zhang, Luke A Summey, Christian Franck, David J Beebe, Hau D Le, Chao Li

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Journal of neuroscience methods, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors.

Nai-Wen LiangDepartment of Biomedical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA.
Ligi MileshDepartment of Surgery, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA.
Karina N ButtramDepartment of Mechanical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA.
Jessica L ParkDepartment of Mechanical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA.
Jing ZhangDepartment of Mechanical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA.
Luke A SummeyDepartment of Mechanical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA.
Christian FranckDepartment of Mechanical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA.
David J BeebeCarbone Cancer Center, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA; Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA. Electronic address: djbeebe@wisc.edu.
Hau D LeCarbone Cancer Center, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA; Department of Surgery, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA. Electronic address: leh@surgery.wisc.edu.
Chao LiCarbone Cancer Center, University of Wisconsin-Madison, 500 Lincoln Drive, Madison, WI 53706, USA; Department of Biomedical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, PA 15213, USA; Department of Chemical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, PA 15213, USA. Electronic address: chaoli3@andrew.cmu.edu.

Funding

UW COMPREHENSIVE CANCER CENTER SUPPORTP30CA014520 · NCI · UNIVERSITY OF WISCONSIN-MADISON · PI Justine Yang Bruce · 1985 to 2026
$142.6M
Under-oil open microfluidic system (UOMS) for studying systemic fungal infectionR01AI154940 · NIAID · UNIVERSITY OF WISCONSIN-MADISON · PI BEEBE, DAVID J, MCCLEAN, MEGAN N · 2021 to 2025
$3.8M
Enhancing Epigenetic Analysis Of Rare Cells With Multi-Phase MicrofluidicsR01CA247479 · NCI · UNIVERSITY OF WISCONSIN-MADISON · PI BEEBE, DAVID J, LANG, JOSHUA MICHAEL · 2020 to 2024
$3.1M
A multiplexed micro scale assay for real time analysis of pediatric immune cell functionU24AI152177 · NIAID · UNIVERSITY OF WISCONSIN-MADISON · PI BEEBE, DAVID J, KERR, SHEENA CATHERINE · 2020 to 2024
$1.8M
Understanding cell migration through microscale in vitro modelsR01EB010039 · NIBIB · UNIVERSITY OF WISCONSIN-MADISON · PI BEEBE, DAVID J, HUTTENLOCHER, ANNA · 2011 to 2014
$1.7M
NCI NIH HHS P30 CA014520NCI NIH HHS R01 CA247479NIAID NIH HHS R01 AI154940NIAID NIH HHS U24 AI152177NIBIB NIH HHS R01 EB010039
6 · The paper itself

Abstract

backgroundCulturing neuronal cells in vitro, especially at smaller scales with reduced media volumes, has been challenging due to the limited proliferation of mature neurons and the inherent high sensitivity of neuronal cells to environmental fluctuations. NEW

methodIn this study, we report a neuronal cell culture method that leverages oil overlay and an autonomously regulated oxygen microenvironment (AROM), in which primary rat cortical cells and human neural progenitor cells (NPCs) were cultured in standard well plates with an oil overlay on top of the media layer. The oil overlay prevents evaporation and achieves in vivo-like oxygen concentrations without the use of glove boxes or hypoxic chambers.

resultsThis oil overlay method achieved > 95 % yield of viable replicates after up to 30 days. Human NPCs cultured under the oil overlay for 15 days exhibited sustained viability without requiring media change. Additionally, oil overlays create a modulated oxygen microenvironment (i.e., AROM) that mimics in vivo conditions, capable of maintaining and restoring optimal oxygen concentrations after disturbances. COMPARISON WITH EXISTING

methodIn contrast, existing method (no-oil controls) resulted in < 20 % yield, low viability for human NPCs (11 % versus 89 % with oil overlay), and oxygen concentrations that returns to ambient levels (21 % oxygen).

conclusionOverall, these results support the oil overlay method as a robust small-scale neuronal cell culture system, offering improved stability and higher yield. The results also underscore the critical role of the oxygen microenvironment in supporting neuronal cell viability, maintenance, and growth.

Indexed as

Cell Culture TechniquesNeural Stem CellsNeuronsOxygenAnimalsCells, CulturedCell SurvivalCerebral CortexHumansRatsOxygenCell culture techniqueHiPSC-derived NPCOil overlayOxygen microenvironment

Identifiers

PMID40466868
PMCPMC12452084

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.