• Journal of Embryo Transfer
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• v.28 no.3
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• pp.281-287
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• 2013

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

• Korean Journal of Agricultural Science
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• v.2 no.1
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• pp.199-231
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• 1975

It is well known fact that antibiotic residues in milk of cows create significant problem for the fermented dairy industry and public health because of inhibition of starter activity and of creation of allergic disease. It can be assumed that antibiotic residues in milk of other aniimals also can create some problems for their infants as in the case of humen. For the above mentioned reasons, present studies were undertaken to determine concentration and duration of antibiotic residues in milk of cows, goats and dogs following intramuscular or intravenous injection and intramammary infusion of penicillin, streptomycin and oxytetracycline at usual dosage. The cylinder-plate method was used for their assay. The results obtained were summerized as follows: 1) Following the intramuscular injection of penicillin, the antibiotic was detected in milk of cows up to 72 hours, in milk of goats 48 hours and in milk of dogs 60 hours of postinjection. The mean peak concentrations were recorded at 12 hours as 0.136 I.U./ml in cows. 6 hours as 0.773 I.U./ml in goats and 3 hours as 1.192 I.U./ml in dogs. 2) Following the intramuscular injection of streptomycin, the antibiotic was detected in milk of cows and goats up to 36 hours and in milk of dogs 24 hours of post-injection. The mean peak concentration were recorded at 6 hours as $0.26{\mu}g/ml$ in cows and at 3 hours in goats and dogs $0.45{\mu}g/ml$ and $0.36{\mu}g/ml$ respectively. 3) Following the intra venous injection of oxytetracycline, the antibiotic was detectable in milk of all the test animals up to 48 hours of postinjection. The mean peak concentrations were recorded at 6 hours as $3.5{\mu}g/ml$ in cows $2.4{\mu}g/ml$ in goats and $2.0{\mu}g/ml$ in dogs respectively. 4) Following intrarnammary infusion of penicillin in amounts of 100,000 I.U. for cows, 20.000 I.U. for goats and 10,000 I.U. for dogs, the penicillin residues in milk of the infused quarter perssisted to 72 hours in cows and 84 hours in goats and dogs. 5) Following intramammary infusion of streptomycin in amount of 500mg for cows, 100mg for goats and 25mg for dogs, the streptomycin residues in milk of the infused quarter persisted to 72 hours in cows and goats and 60 hours in dogs. 6) Following intramammary infusion of oxytetracycline in amount of 500mg for cows, 100mg for goats and 25mg for dogs, the oxytetracycline residues in milk of the infused quarter persisted to 72 hours in cows and 60 hours in goats and dogs. 7) A corelation between the residual antibiotic concentration and milk yield in cows and goats was observed; That is, the lower in the milk production showed a higher the concentration of an antibiotic residues and a longer the time in persistance. 8) Intramammary transfer of the antibiotic from an infused to non infused quarters, in dogs, was observed following the intramammary infusion of penicillin. streptomycin and oxytetracyclne in amounts of 10.000 I.U. 25mg and 25mg respectively. However, no transfer by 100.000 I.U. or 20.000 I.U. of penicillin. 500mg of streptomycin and 100mg of oxytetracyline was observed in cows and goats. 9) In dogs, minimum dosage of antibiotics for transfer fro in treated to untreated quarters following intramammary infusion were 2,500 I.U. of penicillin and 5mg each of streptomycin and oxytetracycline.