Miller Daniel H, Sokol Ethan S, Gupta Piyush B
Whitehead Institute for Biomedical Research, 455 Main Street, Cambridge, MA, 02142, USA.
Department of Biology, Massachusetts Institute of Technology, 31 Ames Street, Cambridge, MA, 02139, USA.
Methods Mol Biol. 2017;1612:139-147. doi: 10.1007/978-1-4939-7021-6_10.
We present a protocol for expanding human mammary tissues from primary patient-derived cells in three-dimensional (3D) cultures. The primary epithelial cells are seeded into 3D hydrogels with defined components, which include both proteins and carbohydrates present in mammary tissue. Over a span of 10-14 days, the seeded cells form mammary tissues with complex ductal-lobular topologies and include luminal and basal cells in the correct orientation, together with cells that stain positively for stem cell markers. In addition to recapitulating key architectural features of human mammary tissue, the expanded tissues also respond to lactogenic hormones including estrogen, progesterone, and prolactin. We anticipate that these cultures will prove useful for studies of mammary development and breast cancer.
我们展示了一种在三维（3D）培养中从原发性患者来源的细胞扩展人类乳腺组织的方案。将原代上皮细胞接种到具有特定成分的3D水凝胶中，这些成分包括乳腺组织中存在的蛋白质和碳水化合物。在10-14天的时间里，接种的细胞形成具有复杂导管-小叶拓扑结构的乳腺组织，包括正确定向的腔上皮细胞和基底细胞，以及对干细胞标志物呈阳性染色的细胞。除了重现人类乳腺组织的关键结构特征外，扩展后的组织还对包括雌激素、孕激素和催乳素在内的泌乳激素有反应。我们预计这些培养物将被证明对乳腺发育和乳腺癌的研究有用。
