Buric Duje, Brisken Cathrin
The Swiss Institute for Experimental Cancer Research (ISREC), Swiss Federal Institute of Technology (EPFL), Station 19, CH-1015, Lausanne, Switzerland.
Methods Mol Biol. 2017;1501:115-129. doi: 10.1007/978-1-4939-6475-8_4.
The mammary gland is the only organ to undergo most of its development after birth and therefore particularly attractive for studying developmental processes. In the mouse, powerful tissue recombination techniques are available that can be elegantly combined with the use of different genetically engineered mouse models to study development and differentiation in vivo.In this chapter, we describe how epithelial intrinsic gene function can by discerned by grafting mammary epithelial cells of different genotypes to wild-type recipients. Either pieces of mammary epithelial tissue or dissociated mammary epithelial cells are isolated from donor mice and subsequently transplanted into recipients whose mammary fat pads were divested of their endogenous epithelium. This is followed by phenotypic characterization of the epithelial outgrowth either by fluorescence stereomicroscopy for the fluorescently marked grafts or carmine alum whole mount for the unmarked epithelia.
乳腺是出生后大部分发育过程才会经历的唯一器官，因此对于研究发育过程特别有吸引力。在小鼠中，可以使用强大的组织重组技术，这些技术可以与不同的基因工程小鼠模型巧妙地结合使用，以研究体内的发育和分化。在本章中，我们描述了如何通过将不同基因型的乳腺上皮细胞移植到野生型受体中来识别上皮内在基因功能。从供体小鼠中分离出乳腺上皮组织块或解离的乳腺上皮细胞，随后将其移植到乳腺脂肪垫已去除内源性上皮的受体中。接下来，通过荧光立体显微镜对荧光标记的移植物进行上皮生长的表型特征分析，或对未标记的上皮进行胭脂红明矾整装标本分析。
