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Pancreatic Tissue Dissection to Isolate Viable Single Cells
Published on: May 26, 2023
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Protocol to isolate human normal and neoplastic pancreatic cells for single-cell omic analyses
Hao Chen1, Junya Peng2, Lulu Liu2
1Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China; State Key Laboratory of Complex Severe and Rare Diseases, Beijing 100730, China.
STAR Protocols
|July 22, 2023
Summary
This study details a protocol for isolating high-quality single cells from human pancreas samples. This method ensures cell viability for advanced single-cell analyses like RNA sequencing.
Area of Science:
- Biotechnology
- Genomics
- Cell Biology
Background:
- High-throughput sequencing demands high-quality cell samples (100-10,000 cells) with 80%-90% viability.
- Efficient single-cell isolation is critical for accurate genomic and transcriptomic profiling.
Purpose of the Study:
- To present a detailed protocol for single-cell isolation from normal and neoplastic human pancreas tissues.
- To ensure high cell viability suitable for downstream single-cell omics applications.
Main Methods:
- Protocol development for human pancreas sample harvesting and procurement.
- Tissue digestion and cell purification techniques for single-cell suspension.
- Application of isolated cells to 10× Genomics single-cell RNA sequencing and single-cell ATAC-seq.
Main Results:
- A robust protocol for obtaining high-quality single-cell suspensions from pancreatic tissue.
- Demonstrated suitability of isolated cells for single-cell RNA sequencing and ATAC-seq.
- Achieved high cell viability meeting platform requirements for single-cell analysis.
Conclusions:
- The presented protocol enables efficient single-cell isolation from human pancreas.
- This method supports advanced single-cell profiling, facilitating research in pancreatic biology and disease.
- The protocol is a valuable resource for researchers utilizing single-cell omics technologies.
