Project description:Bovine mammosphere-derived epithelial cells are a population with reported stem cell-like properties. We assessed these cells using scRNA-seq for markers of pluripotency and other features associated with the effects of their conditioned medium (CM).
Project description:Bovine milk derived cells are a novel population with reported stem cell-like properties. We assessed these cells using scRNA-seq for markers of pluripotency and other features associated with the effects of their conditioned medium (CM).
Project description:Purpose: to detect expression profile of differentially expressed mRNAs during bovine mammary epithelial cells (MAC-T) transfected with miR-375 inhibitor or negative control (NC) inhibitor in vitro. Methods: bovine mammary epithelial cells were transfected with miR-375 inhibitor or negative control (NC) inhibitor to assess the expression profiles of mRNAs using RNA-seq. Results: silencing miR-375 down-regulated and upregulated the expression of 48 and 15 mRNAs, respectively, in bovine mammary epithelial cells. Conclusion: miR-375 silencing dysregulated the expression of 63 mRNAs in bMECs. Also, miR-375 silencing increased the expression of NR4A1 and PTPN5 genes, all anti-inflammatory genes, via the MAPK signaling pathway. Given the negative correlation between miR-375 expression and NR4A1 and PTPN5 genes, miR-375 potentially promotes inflammation in the mammary gland through the MAPK signaling pathway. The findings of this study provide a new perspective of treating mastitis in cows.
Project description:Purpose: to detect expression profile of RNA (lncRNA and circRNA) and elucidate differentially expressed RNAs (DElncRNAs and DEcircRNAs) with potential roles during lipopolysaccharide (LPS)-induced inflammation models of bovine mammary epithelial cells MAC-T in vitro. Methods: bovine mammary epithelial cells MAC-T were exposed to LPS for 0, 6 and 12 hours to assess the expression profiles of RNA (lncRNA and circRNA) using RNA-seq. Results: totally 112 DElncRNAs and 71 DEcircRNAs were screened out at different time points. Functional enrichment analysis on target genes of lncRNAs and host genes of circRNAs indicated that these genes were involve in regulating inflammation-related signaling pathways, including Notch, NF-κB, MAPK, PI3K-Akt, mTOR, MAPK and NOD-like receptor signaling pathway. Conclusion: these differentially expressed RNAs (DElncRNAs and DEcircRNAs) may be involved in the regulation of a variety of immune-related processes including inflammatory responses bovine mammary epithelial cells exposed to LPS via some vital signaling pathways. This study lays a foundation for further research on molecular regulation of bovine mastitis, and also provides a reference for breeding strategies based on molecular markers for mastitis resistance in dairy cows.
Project description:In vitro 2D cultures of intestinal epithelial cells or epithelial cell lines have been widely used to study cell function and host-pathogen interactions in the bovine intestine. However, these cultures lack the cellular diversity encountered in the intestinal epithelium, and the physiological relevance of monocultures of transformed cell lines is uncertain. Little is also known of the factors that influence cell differentiation and homeostasis in the bovine intestinal epithelium, and few cell-specific markers that can distinguish the different intestinal epithelial cell lineages have been reported. Here we describe a simple and reliable procedure to establish in vitro 3D enteroid, or “mini gut”, cultures from bovine small intestinal (ileal) crypts. These enteroids contained a continuous central lumen lined with a single layer of polarized enterocytes, bound by tight junctions with abundant microvilli on their apical surfaces. Histological and transcriptional analyses suggested that the enteroids comprised a mixed population of intestinal epithelial cell lineages including intestinal stem cells, enterocytes, Paneth cells, goblet cells and enteroendocrine cells. We show that bovine enteroids can be successfully maintained long-term through multiple serial passages without observable changes to their growth characteristics, morphology or transcriptome. Furthermore, the bovine enteroids can be cryopreserved and viable cultures recovered from frozen stocks. Our data suggest that these 3D bovine enteroid cultures represent a novel, physiologically-relevant and tractable in vitro system in which epithelial cell differentiation and function, and host-pathogen interactions in the bovine small intestine can be studied.
Project description:In this study, we focused on the changes of mRNA expression in bovine oviduct epithelial cells in vitro co-cultured with embryos during preimplantation development in order to identify genes, which might be involved in embryo-maternal communication. For this purpose, we used large-scale cDNA microarray hybridizations to identify the genes differentially regulated in bovine oviduct epithelial cells (Boec) during in vitro culture. The main objective was to identify genes which are differentially expressed due to the presence or absence of bovine embryos on cell monolayers.