3D differentiation of LUHMES cell line to study recovery and delayed neurotoxic effects

Research output: Research - peer-reviewArticle

Abstract

Current neurotoxicity testing and the study of molecular mechanisms in neurodegeneration in vitro usually focuses on acute exposures to compounds. 3D Lund human mesencephalic (LUHMES) cells allow long-term treatment or pulse exposure in combination with compound washout to study delayed neurotoxic effects as well as recovery and neurodegeneration pathways. In this unit we describe 3D LUHMES culture and characterization. Characterization of the model involves immune cytochemistry, flow cytometry, and qPCR measurements. Studying the delayed effects of compounds is more relevant to human exposures and neurodegenerative diseases with a strong genetic or environmental component. Most assays for molecular endpoints have been developed for monolayer cell culture and therefore need to be adapted for 3D models. In this unit, we further describe toxicological assays for molecular endpoints such as ATP levels, mitochondrial viability, and neurite outgrowth, which have been adapted for use in 3D LUHMES cultures.

LanguageEnglish (US)
JournalCurrent Protocols in Toxicology
Volume2017
DOIs
StatePublished - 2017

Fingerprint

Assays
Cells
Recovery
Cell Line
Neurodegenerative diseases
Flow cytometry
Cell culture
Monolayers
Adenosine Triphosphate
Testing
Endpoint Determination
Histocytochemistry
Neurodegenerative Diseases
Toxicology
Flow Cytometry
Cell Culture Techniques
Therapeutics
Neuronal Outgrowth
In Vitro Techniques

Keywords

  • 3D culture
  • Delayed cellular response
  • LUHMES
  • Neurotoxicity
  • Washout

ASJC Scopus subject areas

  • Toxicology

Cite this

@article{7c24754ff8b64925857f0038d4c285d7,
title = "3D differentiation of LUHMES cell line to study recovery and delayed neurotoxic effects",
abstract = "Current neurotoxicity testing and the study of molecular mechanisms in neurodegeneration in vitro usually focuses on acute exposures to compounds. 3D Lund human mesencephalic (LUHMES) cells allow long-term treatment or pulse exposure in combination with compound washout to study delayed neurotoxic effects as well as recovery and neurodegeneration pathways. In this unit we describe 3D LUHMES culture and characterization. Characterization of the model involves immune cytochemistry, flow cytometry, and qPCR measurements. Studying the delayed effects of compounds is more relevant to human exposures and neurodegenerative diseases with a strong genetic or environmental component. Most assays for molecular endpoints have been developed for monolayer cell culture and therefore need to be adapted for 3D models. In this unit, we further describe toxicological assays for molecular endpoints such as ATP levels, mitochondrial viability, and neurite outgrowth, which have been adapted for use in 3D LUHMES cultures.",
keywords = "3D culture, Delayed cellular response, LUHMES, Neurotoxicity, Washout",
author = "Georgina Harris and Helena Hogberg and Thomas Hartung and Lena Smirnova",
year = "2017",
doi = "10.1002/cptx.29",
volume = "2017",
journal = "Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]",
issn = "1934-9254",
publisher = "John Wiley and Sons Inc.",

}

TY - JOUR

T1 - 3D differentiation of LUHMES cell line to study recovery and delayed neurotoxic effects

AU - Harris,Georgina

AU - Hogberg,Helena

AU - Hartung,Thomas

AU - Smirnova,Lena

PY - 2017

Y1 - 2017

N2 - Current neurotoxicity testing and the study of molecular mechanisms in neurodegeneration in vitro usually focuses on acute exposures to compounds. 3D Lund human mesencephalic (LUHMES) cells allow long-term treatment or pulse exposure in combination with compound washout to study delayed neurotoxic effects as well as recovery and neurodegeneration pathways. In this unit we describe 3D LUHMES culture and characterization. Characterization of the model involves immune cytochemistry, flow cytometry, and qPCR measurements. Studying the delayed effects of compounds is more relevant to human exposures and neurodegenerative diseases with a strong genetic or environmental component. Most assays for molecular endpoints have been developed for monolayer cell culture and therefore need to be adapted for 3D models. In this unit, we further describe toxicological assays for molecular endpoints such as ATP levels, mitochondrial viability, and neurite outgrowth, which have been adapted for use in 3D LUHMES cultures.

AB - Current neurotoxicity testing and the study of molecular mechanisms in neurodegeneration in vitro usually focuses on acute exposures to compounds. 3D Lund human mesencephalic (LUHMES) cells allow long-term treatment or pulse exposure in combination with compound washout to study delayed neurotoxic effects as well as recovery and neurodegeneration pathways. In this unit we describe 3D LUHMES culture and characterization. Characterization of the model involves immune cytochemistry, flow cytometry, and qPCR measurements. Studying the delayed effects of compounds is more relevant to human exposures and neurodegenerative diseases with a strong genetic or environmental component. Most assays for molecular endpoints have been developed for monolayer cell culture and therefore need to be adapted for 3D models. In this unit, we further describe toxicological assays for molecular endpoints such as ATP levels, mitochondrial viability, and neurite outgrowth, which have been adapted for use in 3D LUHMES cultures.

KW - 3D culture

KW - Delayed cellular response

KW - LUHMES

KW - Neurotoxicity

KW - Washout

UR - http://www.scopus.com/inward/record.url?scp=85027010892&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85027010892&partnerID=8YFLogxK

U2 - 10.1002/cptx.29

DO - 10.1002/cptx.29

M3 - Article

VL - 2017

JO - Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]

T2 - Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]

JF - Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]

SN - 1934-9254

ER -