从鼠白色脂肪组织中分离细胞核用于单细胞基因组学。

Center for Functional Genomics and Tissue Plasticity, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M 5230, Denmark.

STAR Protoc. 2021 Jun 17;2(3):100612. doi: 10.1016/j.xpro.2021.100612. eCollection 2021 Sep 17.

Lipid-filled adipocytes are incompatible with droplet-based single-cell methods, such as 10x Genomics-based technology, thus restricting droplet-based single-cell analyses of adipose tissues to the stromal vascular fraction. To overcome this limitation and obtain cellular and molecular insight into adipose tissue composition and plasticity, single-nucleus sequencing-based technologies can be applied. Here, we provide an optimized protocol for nuclei isolation from mouse adipose tissues suitable for single-nucleus RNA sequencing. This allows for transcriptomic profiling of the entire adipose tissue at single-cell resolution. For complete details on the use of this protocol, please refer to Sárvári et al., 2021.

富含脂滴的脂肪细胞与基于液滴的单细胞方法（如 10x Genomics 技术）不兼容，因此限制了基于液滴的单细胞分析应用于脂肪组织的基质血管部分。为了克服这一限制，并深入了解脂肪组织的组成和可塑性，可应用基于单个核测序的技术。在这里，我们提供了一种从小鼠脂肪组织中分离核的优化方案，该方案适用于单细胞 RNA 测序。这使得能够以单细胞分辨率对整个脂肪组织进行转录组分析。如需详细了解本方案的使用方法，请参考 Sárvári 等人，2021 年。