Department 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.
State Key Laboratory of Complex Severe and Rare Diseases, Beijing 100730, China; Medical Research Center, Peking Union Medical College Hospital, Peking Union Medical College, Beijing 100730, China.
STAR Protoc. 2023 Sep 15;4(3):102464. doi: 10.1016/j.xpro.2023.102464. Epub 2023 Jul 24.
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) and Li et al. (2021)..
高通量测序需要高质量的输入细胞样本，每个样本的细胞数为 100-10000 个，细胞活性至少为 80%-90%，具体要求取决于应用的平台。本方案介绍了从正常和肿瘤人类胰腺中分离单细胞的方法。我们描述了样本采集、获取、组织消化和细胞纯化的步骤。随后，分离的细胞可进一步用于单细胞分析，例如使用 10× Genomics 平台进行单细胞 RNA 测序和单细胞 ATAC-seq。如需了解本方案的详细使用和执行方法，请参考 Peng 等人（2019 年）和 Li 等人（2021 年）的文献。
