Ren Gang, Sharma Vandana, Letson Joshua, Walia Yashna, Fernando Veani, Furuta Saori
Department of Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, 3000 Arlington Ave. Toledo, OH 43614, USA.
Bio Protoc. 2019 Oct 5;9(19):e3392. doi: 10.21769/BioProtoc.3392.
Co-culture systems utilizing reconstituted or synthetic extracellular matrix (ECM) and micropatterning techniques have enabled the reconstruction of surface epithelial tissues. This technique has been utilized in the regeneration, disease modeling and drug screening of the surface epithelia, such as the skin and esophagus. On the other hand, the reconstruction of glandular epithelia would require more intricate ECM organizations. Here we describe a protocol for a novel three-dimensional organotypic co-culture system for the reconstruction of mammary glands that utilizes the discontinuous ECM. In this technique, primary mammary fibroblasts first establish a layer of the connective tissue rich in collagen I. Then, mammary epithelial cells form acinar structures, the functional glandular units, within the laminin-rich basement membrane embedded in the connective tissue. This method allows for the regeneration of the -like architecture of mammary glands and could be utilized for monitoring the real-time response of mammary glands to drug treatment.
利用重组或合成细胞外基质（ECM）以及微图案化技术的共培养系统已能够重建表面上皮组织。该技术已被用于皮肤和食道等表面上皮的再生、疾病建模和药物筛选。另一方面，腺上皮的重建需要更复杂的ECM组织。在此，我们描述了一种利用不连续ECM重建乳腺的新型三维器官型共培养系统的方案。在该技术中，原代乳腺成纤维细胞首先形成一层富含I型胶原的结缔组织。然后，乳腺上皮细胞在嵌入结缔组织的富含层粘连蛋白的基底膜内形成腺泡结构，即功能性腺单位。该方法能够再生类似乳腺的结构，并可用于监测乳腺对药物治疗的实时反应。
