It is well known that embryos cultured in a group can create a microenvironment through secretion of autocrine and paracrine factors that can support and improve the embryos' development when compared to the embryos cultured individually. In this study, we used a co-culture system for paracrine communication between different kinds of embryos. The results showed that co-culture of porcine parthenogenetic (PA) embryos significantly improved the in vitro development of cloned (nuclear transfer, NT) embryos. To reveal the possible mechanism of communication between the two groups, we isolated exosomes/microvesicles (EXs/MVs) from the PA embryos conditioned medium (PA-CM) through differential centrifugation and identified them through transmission electron microscope and immunoflourescence against exosomal/membrane marker CD9. Furthermore, these EXs/MVs were found to contain mRNA of pluripotency genes (Oct4, Sox2, Klf4, c-Myc, and Nanog), and the PKH67-labeled EXs/MVs could be internalized by the NT embryos. The current study demonstrates that cloned embryos' developmental competence can be improved through co-culturing with PA embryos and revealed, for the first time, that in vitro-produced embryos can secrete EXs/MVs as a possible communication tool within their microenvironment. Moreover, it provides a new paradigm for embryo-to-embryo communication in vitro.

Cross talk, mediated by exosomes, between normal stem cells and cancer stem cells (CSCs) in the tumor microenvironment has been given less attention so far. In addition, no publications are available in the literature that address the in vivo impact of exosomes derived from CSCs and mesenchymal stem cells (MSCs) on progression of long-term hepatocellular carcinoma (HCC). Herein, we hypothesized that transfer of exosomes among the cells in the HCC microenvironment could either induce or inhibit tumor growth and metastasis depending on their source. To check this hypothesis, we investigated the effect of exosomes coming from two different stem cell populations, hepatic CSCs and bone marrow (BM) MSCs, on progression of long-term DEN-induced HCC in rats and the involved underlying mechanisms. CSCs-exosomes induced a significant increase in liver relative weight and serum levels of cancer markers (AFP and GGT) and liver enzymes (ALT, AST, and ALP), intensive immunostaining for the HCC marker GST-P, and an increased number and area of tumor nodules as compared to HCC rats injected by PBS. CSCs-exosomes also decreased apoptosis (marked by downregulation of Bax and p53 and upregulation of Bcl2, and increased immunostaining of PCNA), increased angiogenetic activity (revealed by upregulation of VEGF), enhanced metastasis and invasiveness (indicated by upregulation of P13K and ERK proteins and their downstream target MMP9 and downregulation of TIMP1), and induced epithelial mesenchymal transition (marked by increased serum and hepatic level of TGFβ1 mRNA and protein). Notably, CSCs-exosomes also elevated HCC exosomal microRNA (miR) 21, exosomal long noncoding (lnc) RNA Tuc339, lncHEIH, and the HCC lncHOTAIR and decreased liver miR122 and HCC miRs (miR148a, miR16, and miR125b). All these cellular, functional, and molecular changes were reversed following injection of BM-MSCs-exosomes. However, both CSCs- and MSCs-exosomes failed to change the elevated oxidative stress or the inhibited antioxidant activities induced by HCC. Collectively, our results revealed a tumor stimulatory effect (induction of tumor growth, progression, and metastasis) for exosomes derived from CSCs and an inhibitory effect for exosomes derived from MSCs. These results provide valuable insight on the effect of CSCs- and MSCs-exosomes on HCC growth and progression in vivo, which may be helpful to understand the mechanism of HCC development.

Objective The current study aimed to replace soybean oil in broiler diets with linseed oil, which is rich in omega-3 fatty acid supplemented with pomegranate peel extract (PPE) and measured its effect on broiler performance, carcass traits, lipid profile, as well as fatty acids composition, phenols and flavonoids content of broiler muscles and immunity of broiler chicks. Methods A total of 300 1-day-old Cobb chicks were randomly allotted into six experimental groups, T1 fed on basal diet with soybean oil without any additives, T2 fed on basal diet with soybean oil with addition of 0.5 g/kg diet PPE, T3 fed on fed on basal diet with soybean oil with addition of 1 g/kg diet PPE, T4 fed on basal diet with linseed oil without any additives, T5 fed on basal diet with linseed oil with addition of 0.5 g/kg diet PPE and T6 fed on basal diet with linseed oil with addition of 1 g/kg diet PPE. The PPE supplementation with 0.05% improved final body weight with either soybean oil ration or linseed oil ration. Results The PPE improved carcass dressing percentage in comparison with the control groups. Body fat levels decreased with increasing PPE levels, especially with a linseed oil diet. Replacing soybean oil with linseed oil decreased the total cholesterol and triacylglycerol levels in broiler serum. The PPE supplementation decreased serum total cholesterol levels and increased high-density lipoprotein cholesterol levels. The content of the breast muscle alpha linolenic acid improved after replacement of soybean oil with linseed oil in broiler diets. PPE supplementation increased the phenol and flavonoid content in broiler meat and increased lysozyme activity. Conclusion Replacing soybean oil with linseed oil in broiler diets with the addition of PPE enriched muscle meat with omega-3 fatty acids and antioxidants and improved broiler immunity and their serum lipid profile.