Microarray expression profiling of distinct subsets of mouse mammary epithelial cells. Mus musculus

The purpose of this microarray experiment was to obtain reference gene expression patterns of a number of epithelial cell populations [mammary stem cells (MASC), luminal progenitors (LP), alveolar luminal stem/progenitor cells (WC virgin-these are mammary epithelial cells genetically marked by Wap-Cre in virgin females), mature luminal cells (ML, mainly represent ductal luminal cells in virgin females), and alveolar luminal cells (WC preg – these are alveolar cells genetically marked by Wap-Cre during mid-gestation)] present in the mammary gland of wildtype adult mice on a C57BL6 genetic background. Overall design: For the isolation of RNA from mammary stem cells (MASC, Lin-CD24+CD29hi), luminal progenitors (LP, Lin-CD24hiCD29+CD61+), and mature luminal cells (ML, lin-CD24hiCD29+CD61-), the thoracic and inguinal mammary glands from 3 adult virgin female mice were harvested, minced and digested into a single cell suspension. Form each of these 3 single cell suspensions, the above populations were sorted by FACS. For alveolar luminal stem/progenitor cells and alveolar luminal cells, Lin-YFP+ mammary epithelial cells were isolated from virgin or midgestation mice genetically marked by Wap-Cre;R26Y. R26Y is a conditional YFP reporter that would be turned on upon Cre-mediated recombination.

本微阵列实验（microarray experiment）旨在获取C57BL6遗传背景下野生型成年小鼠乳腺内多种上皮细胞群的参考基因表达谱，所涉细胞群包括：乳腺干细胞（mammary stem cells, MASC）、腔系祖细胞（luminal progenitors, LP）、肺泡腔系干/祖细胞（WC virgin——即未交配雌性小鼠中经Wap-Cre遗传标记的乳腺上皮细胞）、成熟腔系细胞（mature luminal cells, ML，主要代表未交配雌性小鼠的导管腔系细胞）以及肺泡腔系细胞（WC preg——即妊娠中期经Wap-Cre遗传标记的肺泡细胞）。
实验总体设计如下：
针对乳腺干细胞（MASC, Lin⁻CD24⁺CD29^hi）、腔系祖细胞（LP, Lin⁻CD24^hiCD29⁺CD61⁺）与成熟腔系细胞（ML, Lin⁻CD24^hiCD29⁺CD61⁻）的RNA提取：收集3只成年未交配雌性小鼠的胸部与腹股沟乳腺组织，将其剪碎并消化为单细胞悬液；随后通过荧光激活细胞分选（Fluorescence Activated Cell Sorting, FACS）从这3份单细胞悬液中分别分选得到上述细胞群。
针对肺泡腔系干/祖细胞与肺泡腔系细胞：从经Wap-Cre;R26Y遗传标记的未交配或妊娠中期小鼠体内分离Lin⁻YFP⁺乳腺上皮细胞。其中R26Y为条件性黄色荧光蛋白（Yellow Fluorescent Protein, YFP）报告基因，可在Cre介导的重组作用下被激活。