This study aimed at investigating whether a porcine follicular fluid (pFF) supplementation positively affects the characteristics of donor cells and the developmental competence of porcine cloned embryos. Ear fibroblast cells (donor cell) from an Massachusetts General Hospital miniature pig were cultured in different culture methods: (1) Dulbecco's modified Eagle's medium (DMEM)+10% FBS (Control); (2) DMEM+0.5% FBS (SS); and (3) DMEM+10% FBS+10% pFF (pFF) for 72 h. In each conditioned medium, the concentrations of 4 amino acids (Thr, Glu, Pro, and Val) in the pFF group were significantly different from those in the control group (p<0.05 or p<0.01). The proliferation of the cells cultured in the SS group was significantly lower than that of the other treatment groups (p<0.01). The population of apoptotic and necrotic cells in the SS group was significantly higher than that of either the control or the pFF group (p<0.01). The number of embryos that cleaved (p<0.05) and developed into blastocysts (p<0.01) in the SS group was significantly lower than that of either the control or the pFF group. Compared to other groups, the blastocysts produced from the donor cells in the pFF group had higher total cells and lower apoptotic cells (p<0.05). It can be concluded that pFF supplementation in the donor cell culture medium positively affects cell death, cell cycle and quality of the cloned blastocyst.

Immunological rejection of the organ grafted onto a primate arises from two antibody mediated processes, hyperacute rejection (HAR) and acute humoral rejection (AHR). Functional ablation of α1,3-galactosyltransferase (GalT) and concurrently overexpression of complement regulatory proteins are known to inhibit HAR and AHR. In previous study, we reported that production of porcine male fibroblasts harboring a MCP expression cassette targeted to GalT locus. In this study, we constructed a different MCP expression cassette, in which the EF1α promoter regulates MCP expression and internal ribosome entry site-mediated neomycin resistance gene expression. Subsequently, this cassette was inserted between the left and the right homologous arms to target exon 9 of the GalT gene. Female fibroblasts were isolated from ear skin of 10 days old miniature pig, and used for nucelofection of the the construct for MCP expression at GalT locus. PCR analysis showed that four clones of forty neomycin resistant clones carry MCP expression cassette at exon 9 of the GalT gene. Two clones analyzed downregulated GalT expression, as determined by quantitative reverse transcriptase polymerase chain reaction. Flow cytometry analysis showed that MCP was efficiently expressed at the cell surface.

The antioxidant, xanthine oxidase, tyrosinase inhibitory activities and polyphenol contents of the fruiting bodies of Pleurotus cornucopae extracted with acetone, hot water and methanol (hereinafter referred to Fr. Ace, Fr. HW and Fr. MeOH). The antioxidant activities in the Fr. Ace, Fr. HW and Fr. MeOH were 93.23%, 89.55% and 92.58%, respectively at the concentration of 2.0 mg/ml. Xanthine oxidase inhibition activity in the Fr. Ace, Fr. HW and Fr. MeOH were 45.84%, 46.50% and 45.60%, respectively at the concentration of 5 mg/ml. Tyrosinase inhibition activity in the Fr. Ace, Fr. HW and Fr. MeOH were 52.11%, 50.12% and 55.81%, respectively at the concentration of 1.0 mg/ml. Total polyphenol contents in the Fr. Ace, Fr. HW and Fr. MeOH were 18.99 mgGAEs/ g, 16.73 mgGAEs/g and 18.66 mgGAEs/g. These experimental results suggested that fruiting bodies of P. cornucopae contained good physio-chemical substances for promoting human health.