Protocol to dissociate and isolate wide-diversity single cells by density gradient centrifugation from human hepatoblastoma tissue.
Cancer
Cell Isolation
RNAseq
Sequencing
Single Cell
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
16 Jul 2023
16 Jul 2023
Historique:
received:
05
02
2023
revised:
22
03
2023
accepted:
19
06
2023
medline:
17
7
2023
pubmed:
17
7
2023
entrez:
17
7
2023
Statut:
aheadofprint
Résumé
Single-cell transcriptome sequencing can characterize various cell types in human liver tissue and facilitate understanding of hepatoblastoma heterogeneity. Here, we present a protocol for isolating hepatocytes and immune cells from human hepatoblastoma samples with high viability. We describe steps for tissue processing, enzymatic digestion, Percoll density gradient separation, cell lysis, cell suspension quality control, and scRNA library construction. We then detail sequencing and data analysis. This protocol is applicable to preparing single-cell suspensions from other human liver tissue samples.
Identifiants
pubmed: 37459235
pii: S2666-1667(23)00416-1
doi: 10.1016/j.xpro.2023.102449
pmc: PMC10511933
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
102449Informations de copyright
Copyright © 2023. Published by Elsevier Inc.
Déclaration de conflit d'intérêts
Declaration of interests The authors declare no competing interests.
Références
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