Project description:Spermatogenesis is an intricate developmental process occurring in testes by which spermatogonial stem cells (SSCs) self-renew and differentiate into mature sperm. The molecular mechanisms for SSC self-renewal and differentiation, while have been well studied in mice, may differ between mice and domestic animals including pigs. To gain knowledge about the molecular mechanisms for porcine SSC self-renewal and differentiation that have to date been poorly understood, here we isolated and enriched primitive spermatogonia from neonatal porcine testes, and exposed the cells to retinoic acid, a direct inducer for spermatogonial differentiation. We then identified that retinoic acid could induce porcine primitive spermatogonial differentiation into leptotene spermatocyte-like cells, which was accompanied by a clear transcriptomic alteration, as revealed by the RNA-sequencing analysis. We also compared retinoic acid-induced in vitro porcine spermatogonial differentiation with the in vivo process, and compared retinoic acid-induced in vitro spermatogonial differentiation between pigs and mice. Furthermore, we analyzed retinoic acid-induced differentially expressed long non-coding RNAs (lncRNAs), and demonstrated that a pig-specific lncRNA, lncRNA-106504875, positively regulated porcine spermatogonial proliferation by targeting the core transcription factor ZBTB16. Taken together, these results would help to elucidate the roles of retinoic acid in porcine spermatogonial differentiation, thereby contributing to further knowledge about the molecular mechanisms underlying porcine SSC development and, in the long run, to optimization of both long-term culture and induced differentiation systems for porcine SSCs.
Project description:The porcine ovarian granulosa cells, when analysed during in vitro cultures, may provide new interesting data on the processes associated with in vitro culture of porcine oocytes, as well as assisted reproduction and in vitro fertilisation processes used in the livestock industry. Porcine ovarian granulosa cells were obtained from 43 crossbred Landrace gilts with a median age of 170 days and weight of 98 kg after slaugheter. The cells were harvested after 0h, 48h, 96h and 144h after begining of culture and submited to RNA isolation and microarray analisys. In this study, we demonstrated the gene expression profile of short term primary cultured porcine ovarian granulosa cells.
Project description:We performed small RNA-seq on Dicer KD and control porcine oocyte, and report endo-siRNAs corresponding to SINE1B are significantly down-regulated by Dicer knockdown and are essential for in vitro maturation of porcine oocyte
Project description:The proper mammalian oocytes maturation is recognized as reaching MII stage and accumulation of mRNA and proteins in cell cytoplasm following fertilization. The proper course of folliculogenesis and oogenesis is orchestrated with morphogenesis significantly influencing further zygote formation and embryos growth. This study was aimed to determinate new transcriptomic markers of porcine oocytes morphogenesis associated with cell maturation capacity. We used microarrays to detail the global programme of gene expression underlying changes before and after in vitro maturation of oocytes in sus scrofa
Project description:The objective of this study was to identify porcine genes which expression was affected in vitro stimulation with LPS from Salmonella typhimurium. Microarray experiment was conducted to reveal genes being significantly differentially expressed in LPS stimulated versus unstimulated porcine alveolar macrophages from two from healthy pigs
Project description:Porcine mammary epithelial cell (PMEC) cultures of three lactating sows were treated with potential mastitis-causing pathogens E. coli and S. aureus in vitro. Subsequently transcriptome profiles were analysed after 3 h and 24 h post-challenge, respectively.
Project description:The most experiments about buccal pouch mucosal cells were investigated in mice and hamster. In these studies we have used porcine buccal pouch mucosal cells cultured primarily in vitro to investigate the expression profile of new molecular markers of mucosal wounding, vascularization and proliferation. In this study, we demonstrated the gene expression profile of long time primary cultured porcine buccal pouch mucosal cells.
Project description:Sus domesticus (pig) are an excellent large mammalian model for comparative studiesdue to their relatively comparable physiology and organ size to humans. The derivation of transgene-free porcine pluripotent stem cells (PiPSCs) will therefore benefit the development of porcine-specific models for regenerative biology and its medical applications. In the past, this effort has been hampered by a lack of understanding of the signaling milieu that stabilizes the porcine pluripotent state in vitro. Here, we report that transgene-free PiPSCs can be efficiently derived from porcine fibroblasts by episomal vectors along with microRNA-302/367 using optimized protocols tailored for this species. Established PiPSCs can robustly differentiate into derivates representing the primary germ layers in vitro and in vivo. Furthermore, the transgene- free PiPSCs preserve intrinsic species-specific developmental timing in culture. This capacity is demonstrated by establishing a porcine in vitro segmentation clock model that, for the first time, displays a specific periodicity at ~ 3.7 hours, a time scale recapitulating in vivo porcine somitogenesis. We therefore propose that these transgene-free PiPSCs can serve as a powerful tool for modeling development and disease, informing both conserved and unique features of mammalian pluripotency and developmental timing mechanisms.
Project description:Recently a lot of experiment were developed based on primary cell culture models, including several species of mammals, organs and tissues. The results from these studies revealed the differences in cells proliferation ability in vitro, which is significantly related to the type of tissue originated cells. In this study, we demonstrated the gene expression profile of long time primary cultured porcine oviductal epithelial cells.
Project description:We investigated for the first time the in vitro response of PRRSV-infected porcine DCs and monocytes to S. suis. We first assessed the effect of PRRSV infection on the phagocytosis and intracellular survival of S. suis by these two cell populations. We then used a genomic approach to compare the gene expression profiles of both type of cells infected with S. suis, with or without a previous infection with PRRSV. Total RNA obtained from porcine monocytes and dendritic cells infected with S. suis, PRRSV, or S. suis & PRRSV. Four replicates in all groups.