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Protocol to isolate human normal and neoplastic pancreatic cells for single-cell omic analyses

The high-throughput sequencing at single-cell resolution requires high-quality samples of inputted 100–10,000 cells with at least 80%–90% viability according to the applied platform. Here, we present a protocol for single-cell isolation from normal and neoplastic human pancreas. We describe steps fo...

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Detalles Bibliográficos
Autores principales: Chen, Hao, Peng, Junya, Liu, Lulu, Huang, Dan, Zhao, Yupei, Wu, Wenming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10382986/
https://www.ncbi.nlm.nih.gov/pubmed/37480562
http://dx.doi.org/10.1016/j.xpro.2023.102464
Descripción
Sumario:The high-throughput sequencing at single-cell resolution requires high-quality samples of inputted 100–10,000 cells with at least 80%–90% viability according to the applied platform. Here, we present a protocol for single-cell isolation from normal and neoplastic human pancreas. We describe steps for sample harvesting, procurement, tissue digestion, and cell purification. Subsequently, isolated cells can be further applied to single-cell profiling, for example, single-cell RNA sequencing and single-cell ATAC-seq using 10× Genomics platform. For complete details on the use and execution of this protocol, please refer to Peng et al. (2019)(1) and Li et al. (2021).(2)