Using intracellular SCGB1A1-sorted, formalin-fixed club cells for successful transcriptomic analysis.

Bronchioles / cytology Flow Cytometry / methods Formaldehyde Gene Expression Profiling / methods Humans Paraffin Embedding RNA, Messenger / isolation & purification Tissue Fixation / methods Transcriptome Uteroglobin / chemistry
Club cells RNA quality SCGB1A1 sorting Transcriptomic analysis

Journal

Biochemical and biophysical research communications
ISSN: 1090-2104
Titre abrégé: Biochem Biophys Res Commun
Pays: United States
ID NLM: 0372516

Informations de publication

Date de publication:
14 05 2022
Historique:
received: 25 02 2022
accepted: 07 03 2022
pubmed: 20 3 2022
medline: 12 4 2022
entrez: 19 3 2022
Statut: ppublish

Résumé

As opposed to surface marker staining, certain cell types can only be recognized by intracellular markers. Intracellular staining for use in cell sorting remains challenging. Fixation and permeabilization steps for intracellular staining and the presence of RNases notably affect preservation of high-quality mRNA. We report the work required for the optimization of a successful protocol for microarray analysis of intracellular target-sorted, formalin-fixed human bronchial club cells. Cells obtained from differentiated air-liquid interface cultures were stained with the most characteristic intracellular markers for club cell (SCGB1A1

Identifiants

DOI: 10.1016/j.bbrc.2022.03.040 PMID: 35305419
pubmed: 35305419
pii: S0006-291X(22)00365-5
doi: 10.1016/j.bbrc.2022.03.040
pii:
doi:

Substances chimiques

RNA, Messenger 0
SCGB1A1 protein, human 0
Formaldehyde 1HG84L3525
Uteroglobin 9060-09-7

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

151-157

Informations de copyright

Copyright © 2022 Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of competing interest The authors declare no competing interests.

Auteurs

Charlotte Vernisse (C)

PhyMedExp, University of Montpellier, INSERM U1046, CNRS UMR9214, France; Medicine Biology Mediterranee, Department of Respiratory Diseases and Addictology, Arnaud de Villeneuve Hospital, CHRU Montpellier, France.

Aurélie Petit (A)

Medicine Biology Mediterranee, Department of Respiratory Diseases and Addictology, Arnaud de Villeneuve Hospital, CHRU Montpellier, France.

Véronique Pantesco (V)

IRMB, University of Montpellier, INSERM, Montpellier University Hospital, France.

Pascal Chanez (P)

Aix-Marseille University, C2VN, AMU - INSERM 1263, INRAE, 1260, Marseille, France; Aix-Marseille University, Department of Respiratory Diseases, Nord Hospital, Marseille, France.

Delphine Gras (D)

Aix-Marseille University, C2VN, AMU - INSERM 1263, INRAE, 1260, Marseille, France; Aix-Marseille University, Department of Respiratory Diseases, Nord Hospital, Marseille, France.

Edouard Tuaillon (E)

Pathogenesis and Control of Chronic Infections, INSERM U1058/EFS, Montpellier University and Laboratory of Virology, Montpellier University Hospital, France.

Christophe Duperray (C)

IRMB, University of Montpellier, INSERM, Montpellier University Hospital, France; MRI, BioCampus Montpellier, CNRS, INSERM, University of Montpellier, France.

Isabelle Vachier (I)

Medicine Biology Mediterranee, Department of Respiratory Diseases and Addictology, Arnaud de Villeneuve Hospital, CHRU Montpellier, France.

Said Assou (S)

IRMB, University of Montpellier, INSERM, Montpellier University Hospital, France.

Arnaud Bourdin (A)

PhyMedExp, University of Montpellier, INSERM U1046, CNRS UMR9214, France; Department of Respiratory Diseases and Addictology, Arnaud de Villeneuve Hospital, Montpellier University Hospital, France. Electronic address: a-bourdin@chu-montpellier.fr.

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Classifications MeSH

questionsmedicales.fr › Appareil respiratoire › Poumon › Bronches › Bronchioles › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Techniques d'investigation › Techniques cytologiques › Cytophotométrie › Cytométrie en flux › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Composés chimiques organiques › Aldéhydes › Formaldéhyde › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Analyse de profil d'expression de gènes › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Techniques d'investigation › Techniques de laboratoire clinique › Techniques histologiques › Techniques de préparation histocytologique › Inclusion de tissu › Inclusion en paraffine › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN messager › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Techniques d'investigation › Techniques de laboratoire clinique › Techniques histologiques › Techniques de préparation histocytologique › Fixation tissulaire › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Transcriptome › Using intracellular SCGB1A1-sorted,...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Sécrétoglobines › Blastokinine › Using intracellular SCGB1A1-sorted,...