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Gene Expression Patterns

Volume 38, December 2020, 119151
Gene Expression Patterns

The mesenchymal property of mouse mammary anlagen repopulating cell population is associated with its stemness

https://doi.org/10.1016/j.gep.2020.119151Get rights and content

Abstract

During early embryogenesis, mammary glands are derived from surface ectoderm and their morphogenesis is controlled by mammary stem cells (MaSCs) and epithelial-mesenchymal transition (EMT). Mammary anlagen stage (E13.5–15.5) is an important stage for fetal mice to achieve EMT dependent mammary morphogenesis. And the characteristics of mammary anlagen repopulating cell population (MaRC) should be identified for understanding its stemness at earlier embryonic stage. Here we quantify and characterize MaSCs proportion at mammary anlagen stage. Compared with adult mouse mammary gland, our data revealed that E14.5 mammary anlagen exhibit higher stem cell activities. Then we purified mammary anlagen cell populations depending on the expression levels of CD24 and CD49f in mouse mammary anlagen, and identified an unique MaRC population (Lin-CD24medCD49f+) by real-time PCR, transplantation and mammosphere forming assays. In addition, by comparing with adult MaSC (Lin-CD24+CD29hi) and differentiated mammary anlagen cells, we find that E14.5 mouse MaRC population exhibit gene expression programs related to mesenchymal properties. To further identify the cell types of E14.5 mouse MaRC population, the expressions of K8, K14, K18, e-cadherin, n-cadherin and vimentin in mammary anlagen Lin-CD24medCD49f + cells were detected by immunofluorescence assay. These findings verified that the undifferentiated E14.5 mouse MaRC population is a heterogeneous population with mesenchymal property, which is associated with cell stemness and mammary duct morphogenesis.

Section snippets

Author statement

Jiazhe Song: Conceptualization, Methodology, Software, Resources, Investigation. Kai Xue: Conceptualization, Data curation, Writing Original draft preparation. Yinhui Liu: Visualization, Investigation. Meng Jia: Formal analysis, Investigation. Shujun Liu: Visualization, Investigation. Lin Lin: Software, Validation. Wenyong Ding: Supervision, Writing- Reviewing and Editing.

The analysis of stem cell activities in fetal mouse mammary anlagen

First, to identify the potential MaSCs distribution of mammary rudiments at earlier embryonic stage (E13.5–15.5), the immunostaining of multiple lineages-associated cytokeratins for fetal mouse mammary anlagen were performed. According to the previous reports, expressions of K14 and K8 have ever been proposed to indicate multipotent cells in the normal mammary gland and in breast cancers (Van Keymeulen et al., 2011; Bai and Rohrschneider, 2012; Spike et al., 2012). The immunostaining showed

Discussion

Morphogenesis of mammary gland in embryonic stage and postnatal life, or the regeneration of functional structure by transplantation are thought to be mainly accomplished by MaSC, which is regulated by different signals, and then producing different types of cells through symmetrical and asymmetrical division. The development of mammary glands originates from the surface ectoderm, and its morphology is also controlled by the interaction of epithelial and mesenchymal substances (Robinson et al.,

Mice

All animal research was approved by Beijing Administration Committee of Laboratory Animals under the leadership of the Beijing Association for Science and Technology. All experiments were carried out according to the guidelines of the American Association for Laboratory Accreditation.

CD-1 and C57 female mice (Vital River Co., Beijing, China) in estrus were maintained and mated in our animal facility. Insemination was verified by the presence of vaginal plugs. Noon of the day following

Funding

This work was supported by National Natural Science Foundation of China (No: 31101717 and 81272429). Science research funding of Lioaning Provincial Education Department (No: LZ2019033, LZ2019062).

Data statement

Data openly available in a public repository that issues datasets with DOIs.

Declaration of competing interest

None.

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