Department of Comparative Biosciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, USA.
Methods Mol Biol. 2022;2471:271-282. doi: 10.1007/978-1-0716-2193-6_15.
Breast cancer is a multifactorial disease with risk factors that are fixed or modifiable. Understanding how these risk factors interact within breast tissue may provide insight into how to improve interventions or chemoprevention strategies to reduce breast cancer incidence. Here we describe methods to utilize breast tissue from patients with defined risk factors undergoing reduction mammoplasty or prophylactic mastectomy to isolate epithelial cells, stromal cells, adipocytes, and macrophages to investigate how risk factors impact distinct cell populations within breast tissue. Following enzymatic digestion of breast tissue, adipocyte-enriched, stromal cell, and epithelial organoid fractions can be isolated. Using antibody-conjugated beads, further cell populations, such as macrophages, can be isolated for molecular analysis. These methods can be adapted to sequentially isolate other cell populations based on specific cell surface markers and are useful for small-sized breast tissue specimens.
乳腺癌是一种多因素疾病，其危险因素可分为固定因素和可改变因素。了解这些危险因素在乳腺组织中的相互作用方式，可能有助于深入了解如何改进干预措施或化学预防策略，以降低乳腺癌的发病率。在这里，我们描述了利用接受缩乳术或预防性乳房切除术的具有明确危险因素的患者的乳腺组织来分离上皮细胞、基质细胞、脂肪细胞和巨噬细胞的方法，以研究危险因素如何影响乳腺组织内不同的细胞群体。在对乳腺组织进行酶消化后，可以分离出富含脂肪细胞、基质细胞和上皮类器官的部分。使用抗体偶联珠，可以进一步分离巨噬细胞等其他细胞群体，用于分子分析。这些方法可以根据特定的细胞表面标志物进行适应性调整，以顺序分离其他细胞群体，对于小型乳腺组织标本非常有用。
