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Direct Reprogramming of Fibroblasts into Functional Mammary Epithelial cells by small molecules (lncRNAseq)

ABSTRACT: Mammary epithelial cells (MECs), the only cell type which could be capable of secreting milk in mammary gland, can be applied for the generation of transgenic mammary cell/gland bioreactor. However, continuously achieving large amount of high-quality goat MECs by non-invasion method, especially prepuberal MECs, in vitro has been a challenge. Here, we show that a small molecules cocktail enables the in vitro generation of chemically-induced mammary epithelial cells (CiMECs) with milk-secreting properties from goat ear fibroblasts. The properties of CiMECs were highly similar to those of naturally isolated MECs. Notably, the CiMECs lineage reprogramming process may partially imitate the MECs developmental process from embryonic to milk-secreting stages. Interestingly, fibroblasts were firstly reversed back to embryonic ectoderm/surface ectoderm-like state. Our findings constitute a substantial step toward further promoting the application of transgenic mammary bioreactors, moreover, also offer a new strategy for obtaining other cell types by reprogrammed fibroblasts.

ORGANISM(S): Capra hircus

PROVIDER: GSE142547 | GEO | 2022/12/20

REPOSITORIES: GEO

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Direct Reprogramming of Fibroblasts into Functional Mammary Epithelial cells by small molecules (miRNAseq)

Project description:Mammary epithelial cells (MECs), the only cell type which could be capable of secreting milk in mammary gland, can be applied for the generation of transgenic mammary cell/gland bioreactor. However, continuously achieving large amount of high-quality goat MECs by non-invasion method, especially prepuberal MECs, in vitro has been a challenge. Here, we show that a small molecules cocktail enables the in vitro generation of chemically-induced mammary epithelial cells (CiMECs) with milk-secreting properties from goat ear fibroblasts. The properties of CiMECs were highly similar to those of naturally isolated MECs. Notably, the CiMECs lineage reprogramming process may partially imitate the MECs developmental process from embryonic to milk-secreting stages. Interestingly, fibroblasts were firstly reversed back to embryonic ectoderm/surface ectoderm-like state. Our findings constitute a substantial step toward further promoting the application of transgenic mammary bioreactors, moreover, also offer a new strategy for obtaining other cell types by reprogrammed fibroblasts.

2022-12-20 | GSE142659 | GEO

Direct Reprogramming of Fibroblasts into Functional Mammary Epithelial cells by small molecules (WGBSseq)

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Isolation and Identification of Mammary Epithelial Cells from Goat Milk

Project description:Mammary epithelial cells (MECs) have strong secretory function, which is an important target cell for studying mammary gland bioreactor. It is also a unique cell with lactation function in mammary gland. It can be used to study cell biological characteristics and molecular biological mechanism in the process of mammary gland growth, development and lactation. At present, mammary epithelial cells are obtained from mammary tissue, and the extraction of tissue block involves surgical anesthesia, which may lead to breast tissue injury and infection. This study aimed to describe the process of isolating and culturing goat mammary epithelial cells (GMECs) from goat milk, and to identify the biological characteristics of GMECs. Our study laid a foundation for the study of the regulation mechanism of mammary gland development and lactation, the improvement of milk quality, the increase of milk production and the establishment of mammary gland bioreactor.

2023-02-20 | GSE145681 | GEO

Liver X Receptor α Activation Enhances Cholesterol Secretion in Lactating Mammary Epithelium

Project description:We profiled differential gene expression in vivo between pregnant day 14 (secretory differentiation) and lactation day 4 (established secretory activation) using isolated mouse mammary epithelial cells depleted of the mammary adipocytes. Liver-X-Receptors are ligand-dependent transcription factors activated by cholesterol metabolites. These receptors induce a suite of target genes required for de novo synthesis of triglycerides and cholesterol transport in many tissues. Two different isoforms -LXRα and LXRβ- have been well characterized in liver, adipocytes, macrophages and intestinal epithelium among others, but their contribution to cholesterol and fatty acid efflux in the lactating mammary gland is poorly understood. We hypothesize that LXR regulates lipogenesis during milk fat production in lactation. Global mRNA analysis of mouse mammary epithelial cells (MECs) revealed multiple LXR/RXR targets are upregulated sharply at the onset of lactation compared to mid-pregnancy. LXRα is the primary isoform and its protein levels increase throughout lactation in MECs. The LXR agonist GW3965 markedly induced several genes involved in cholesterol transport and lipogenesis and enhanced cytoplasmic lipid droplet accumulation in the HC11 MEC cell line. Importantly, in vivo pharmacological activation of LXR increased the milk cholesterol percentage and induced Srebp1c and Abca7 expression in MECs. Cumulatively, our findings identify LXRα as an important regulator of cholesterol incorporation into the milk through key nodes of de novo lipogenesis, suggesting a potential therapeutic target in women with difficulty initiating lactation.

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Discovery and Comparative Transcriptome Profiling of Dairy Goat MicroRNAs from Dry Period and Peak Lactation Mammary Gland Tissues

Project description:In this study, two small RNA libraries were constructed using dry period and peak lactation dairy goat mammary gland tissues and sequenced by the Illumina Solexa high-throughput sequencing system. A total of 346 conserved and 95 novel miRNAs were identified in the dairy goat. The expression of miRNAs was confirmed by qRT-PCR in nine tissues and the mammary gland during development cycles. In addition, several candidate miRNAs that may be involved in mammary gland development and lactation were found by the comparison of miRNA expression profiles among different tissue and developmental stages of the mammary gland. This study provides the identification and profile of miRNAs related to the biology of the mammary gland in the dairy goat. The identification of these miRNAs could contribute to understanding the molecular mechanisms of lactation physiology and the development of the mammary gland in the dairy goat.

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Project description:Animal nutrition considerably affects milk composition that influences its nutritional quality. Milk component synthesis and secretion by the mammary gland involve the expression of a large number of genes whose nutritional regulation remains poorly defined. In this study, 16 lactating goats received 4 experimental diets differing in either forage to concentrate ratio (high forage, HF, or low forage, LF) supplemented, or not, with lipids (whole rapeseeds, RS, or sunflower oil, SO) in a 4 x 4 Latin Square design. To investigate the pathways regulated by nutrition, we examined the effect of these diets on the expression of approximately 8400 genes in caprine mammary gland using a bovine oligonucleotide microarray. Due to the limited quantity of mammary RNA available, equal amounts of total RNA sample from mammary gland of each goat belonging to the same Latin Square group were mixed together before labeling. Each mammary pooled sample (4 by dietary treatment) was then co-hybridized with a standard reference corresponding to a mixture of purified total RNA from several caprine tissues. Each hybridization was repeated in a dye-swap manner for a total of 32 slides (8 slides and 4 independent comparisons by dietary treatment).

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Expression data from sorted control and Pygo2-null mouse mammary stem/basal cells

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