This study investigated the effect of variation in the number of somaticcell- cloned embryos and their developmental stage at transfer on pregnancy, as well as the influence of the estrus status of recipient pigs on in vivo development of cloned porcine embryos after embryo transfer. For somatic cell nuclear transfer (SCNT), fibroblast cells were obtained from a male porcine fetus. Recipient oocytes were collected from prepubertal gilts at a local abattoir and then cultured. After SCNT, reconstructed embryos of different numbers and developmental stages were transferred into recipient pigs. The developmental stage of the cloned embryos and the number of transferred embryos per surrogate showed no significant differences in terms of the resulting cloning efficiency. However, the pregnancy rate improved gradually as the number of transferred cloned embryos was increased from 100- 150 or 151-200 to 201-300 per recipient. In pre-, peri-, and post-ovulation stages, pregnancy rates of 28.6%, 41.8%, and 67.6% and 16, 52, and 74 offspring were recorded, respectively. The number of cloned embryos and estrus status of the recipient pig at the time of transfer of the cloned embryo affect the efficiency of pig production; therefore, these variables should be particularly considered in order to increase the efficiency of somatic cell pig cloning.

Porcine parvovirus 2 (PPV2) was recently detected in the Republic of Korea. This paper reports two near-complete genome sequences of PPV2 identified for the first time in the lung tissue of aborted pig fetuses.

This study examined the spatial autocorrelation of the 2016 foot and mouth disease (FMD) outbreaks in South Chungcheong to determine the association between the disease epidemics and pig farm vehicle movements. Two spatial autocorrelation testing methods were used: Moran’s I and Getis-Ord G statistics. The Moran’s I statistic for the FMD outbreak areas was -0.239, and its p-value was less than 0.007. The median Getis-Ord G statistic for the FMD outbreak areas was -0.323. The results indicated that the geographical distribution of the FMD outbreak areas was not spatially homogeneous. The spatial autocorrelation of the 2016 FMD epidemics was considered by applying a geographical weighted Poisson regression (GWPR) model in the analysis, in which pig farm vehicle movements were used as risk factors for the 2016 FMD epidemics. The number of FMD-infected farms per second-level administrative province (si or gun) was used as a dependent variable. The number of farm vehicle movements within the province (within variable), from one province to other provinces (outbound variable), or from other provinces to one province (inbound variable), were included as independent variables in the GWPR model. The results of the GWPR model were as follows. The estimated median coefficient of the log-transformed within variable, the log-transformed outbound variable, and log-transformed inbound variable were -0.000, 0.010, and -0.009, respectively. The optimal bandwidth for the GWPR model was 80.49, and the AIC score was 89.35. The results showed that the GWPR model would provide an understanding of the relationship between the 2016 FMD epidemics and pig farm vehicle movements

As a preclinical study, many researchers have been attempted to convert the porcine PSCs into several differentiated cells with transplantation of the differentiated cells into the pigs. Here, we attempted to derive neuronal progenitor cells from pig embryonic germ cells (EGCs). As a result, neuronal progenitor cells could be derived directly from pig embryonic germ cells through the serum-free floating culture of EB-like aggregates (SFEB) method. Treating retinoic acid was more efficient for inducing neuronal lineages from EGCs rather than inhibiting SMAD signaling. The differentiated cells expressed neuronal markers such as PAX6, NESTIN, and SOX1 as determined by qRT-PCR and immunostaining. These data indicated that pig EGCs could provide valid models for human therapy. Finally, it is suggested that developing transgenic pig for disease models as well as differentiation methods will provide basic preclinical data for human regenerative medicine and lead to the success of stem cell therapy.

한국의 경제 발전에 따라 양돈 사업의 규모가 커지고 대단지화 되면서, 생산성 향상과 높은 품질을 충족시키기 위해 축사시설의 체계적인 관리가 요구되고 있다. 그러나 효율성을 위해 밀집된 구조에서는 종종 여름철에 공기의 질이 나빠지고 온도가 높아지는 등의 문제를 일으킬 수 있다. 돼지는 저산소 및 고온에 매우 취약하기 때문에 생산성을 높이기 위한 환기 유지는 매우 중요하다. 이에 본 연구에서는 전산유체역학(CFD, computational fluid dynamics)을 이용하여 환기 시스템의 문제점 및 개선 사항을 분석하는 방법을 제안하였다. 유한체적법(FVM, finite volume method) 기반의 fluent 프로그램이 사용되었고 난류 모델로 RNG standard k-ε 모델을 사용하였다. 환기 시스템의 효율적인 설계를 위해 실제 단층의 단순 구조 양돈사와 다층의 대형 양돈사를 대상으로 다종물질이송(multispecies transport) 해석기법을 이용하여 환기 효율 및 성능을 분석하였으며, 개선된 시스템의 유동순환이 원활하게 이루어짐을 확인하였다.

오늘날 양돈의 사양이 증가 함에 따라 부존사료 자원으로서 활용가치가 높은 물질에 항생제를 첨가 하지 않고 친환경적으로 제조된 가축용 사료를 개발하여, 기능성 친환경 돈육 및 가공품을 생산하는 것이 중요하다. 이에 따라 본 연구에서는 항암, 항염, 항산화 등 다양한 생리활성 물질이 다량 함유되어 있는 복분자(Rubus occidentalis, RO) 부산물을 양돈사료의 원료로 확립하고자 버크셔 돼지를 대상으로 복분자 발효사료(Rubus occidentalis fermented fodder, ROFF)의 혈액 내 영양 운반인자와 항산화 효능을 연구하기 위해 수행하였다. 시험구는 일반사료(대조구)에 ROFF를 0.3% 첨가하였고, 발효사료의 효능을 확인 하기 위해 각 체중에 따른 버크셔 거세돈 및 암퇘지에게 43~73일간 급여하여 효능을 확인 하였다. ROFF의 복합적인 효능은 혈액 생화학, 철분 및 항산화 분석을 통해 확인하였다. 그 결과 ROFF의 섭취로 인해 거세 자돈 및 육성돈, 암컷 자돈, 육성돈, 및 110-150 kg 암컷 비육돈 에서 total cholesterol (TC), LDL-cholesterol (LDL-C), HDL-cholesterol (HDL-C) 모두 감소하거나 HDL-C이 증가하는 경향으로 보아 영양생리학적으로 개선되는 경향을 확인하였다. 암컷 비육돈에서는 생화학적 수치가 모두 증가하는 경향으로 보아 임신 가능성이 있는 비육기의 경우 이러한 수치 향상으로 자돈의 생산을 위한 원활한 영양 공급이 예상되며, 건강한 자돈의 생산에 도움이 될 것으로 판단된다. Transferrin (TFE) 함량은 거세돈에서는 변화가 크지 않았으며, 암컷 육성돈 및 110-150 kg 암컷 비육돈에서 ROFF 섭취로 인해 증가하는 경향이 나타남에 따라 ROFF가 철분 결핍으로 인한 부정적인 영향들을 최소화 할 수 있을 것이다. Glutathione peroxidase1 (GPx1)분석 결과 거세 비육 돈 및 110-150 kg 암컷 비육돈에서 ROFF의 섭취로 인해 GPx1 활성이 유의적이지는 않지만 크게 증 가한 것으로 보아 ROFF가 항산화능을 향상시키는 것으로 판단된다. 본 연구의 결과들을 종합할 때, ROFF의 급여가 전체적으로 시험 개체의 개선 효능에는 영향이 크지 않으나 RO의 함량을 고려 할 때 매우 긍정적인 원료사료 중 하나라 할 수 있으며, 특히 ROFF가 거세돈 보다는 암컷 버크셔의 영양 운 반 및 철분 함량 등에 긍정적인 영향을 줄 수 있을 것으로 판단된다.

상업적 영농을 하는 양돈농가는 돼지의 생산비를 줄이면서 소비자들이 선호하는 고품질의 고기를 생산하는 것이 소득증 대에 있어서 무엇보다 중요한 일이다. 이에 농림축산식품부는 돼지도체를 1+, 1, 2 및 등외등급으로 판정하고, 그 결과에 따라 농가수취가격을 차등화함으로써 고품질 돼지고기 생산을 촉진하기 위해서 돼지고기 도체등급제도를 시행하고 있으며, 돼지도체의 체중과 등지방두께를 기준으로 1차 등급판정을 한 다. 가장 높은 등급인 1+ 등급은 돼지의 탕박 기준으로 도체 중이 83-93 kg, 등지방두께가 17~25 mm이다. 축산물등급판정 통계연보에 의하면, 2018년 경락단가가 가장 높은 평균 도체 중량은 암퇘지가 83.5 kg, 거세돼지가 77.8 kg이었으며, 평균 등지방두께는 암퇘지가 19.2 mm, 거세돼지가 20.3 mm인 것으로 조사되었다. 본 연구는 거세돼지와 암퇘지에 대한 분산분석, 육량변수와 육질변수 사이의 개별 상관관계, 그리고 육량변수세트와 육질 변수세트 사이의 정준상관분석을 수행하였다. 먼저 분산분석의 결과는 다음과 같이 요약할 수 있다. 첫째, 돼지의 육량 및 육질특성변수들에 있어서 성별 차이를 분석한 결과는 등지방두께는 거세돼지가 암퇘지보다 0.14㎜ 두꺼웠다(P<0.05). 둘째, 육즙감량과 가열감량은 거세돼지가 암퇘지보다 많았다 (P<0.05). 셋째, 보수력, 콜라겐함량, 지방함량, 수분함량, 단백질함량 및 전단가는 암퇘지가 거세돼지보다 높았다 (P<0.05). 육량특성과 육질특성 사이의 상관분석 결과는 다음과 같이 요약할 수 있다. 첫째, 도체중이 무거울수록 등지방두께가 두껍고, 육즙감량이 많이 생기지만, 가열감량은 줄어드는 관계를 갖는다 (P<0.01). 둘째, 등지방두께가 두꺼울수록 지방함량이 늘어나고, 육즙 감량이 많이 생기지만, 콜라겐함량, 수분함량, 단백질함량 및 가열감량과는 음(-)의 상관관계를 갖는 것으로 나타났다 (P<0.01). 셋째, 수분함량과 지방함량의 상관관계가 가장 높았으며 (0.793), 이어서 단백질함량과 지방함량의 상관관계가 높았지만 (0.634), 이들 함량 간에서는 서로 음(-)의 관계를 보이고 있다 (P<0.01). 마지막으로 정준상관분석의 결과는 다음과 같이 요약할 수 있다. 첫째, 육량특성변수 짝과 육질특성변수 짝 사이에 유의한 관계가 있는 것으로 나타났다 (P<0.01). 둘째, 등지방두께(BFT)가 높으면 지방함량과 육즙감량은 높은 반면, 수분함량, 단백질함량, 가열감량, 콜라겐함량, 전단가 및 보수력이 낮은 것으로 나타났다. 이와 같은 결과에 따르면, 소비자들은 지방함량이 많으면서 수분손실이 많은 돼지고기와 단백질함량과 콜라겐함량이 적은 돼지고기는 기피할 수 있다는 점을 간과하여서는 안 될 것이다. 특히 국내 돼지도체등급은 1차적으로 돼지도체중량과 등지방두께를 평가한 이후, 육질항목을 평가하여 최종등급이 판정 되고 있기 때문에 우선적으로 육량지수인 도체중량과 등지방 두께를 적절하게 관리하는 것이 소비자의 요구를 충족하면서 농가의 생산비 절감 및 농가수취가격상승에 의한 소득증대로 이어지는 중요한 요인이 된다는 사실에 주목할 필요가 있다.

Stem cells are progenitor cells that are capable of self-renewal and differentiation into various cells. Especially, pluripotent stem cells (PSCs) have in vivo and in vitro differentiation capacity into three germ layers and can proliferate infinitely. The differentiation ability of PSCs can be applied for regenerative medicine and tissue engineering. In domestic animals, their PSCs have a potential for preclinical therapy as well as the production of transgenic animals and agricultural usage such as cultured meat. Among several domestic animals, a pig is considered as an ideal model for biomedical and agricultural purposes mentioned above. In this reason, studies for pig PSCs including embryonic stem cells (ESCs), embryonic germ cells (EGCs) and induced pluripotent stem cells (iPSCs) have been conducted for decades. Therefore, this review will discuss the history of PSCs derived from various origins and recent progress in pig PSC research field.

Tight junctions are constituents of the blood–epididymis barrier that play roles in regulating the unidirectional transcellular transport of ions, water, and solutes to maintain optimal conditions for sperm maturation and storage. Claudin 1 (Cldn1) and 4 (Cldn4) are known as tight junction proteins and are expressed in the basolateral membranes as well as tight junctions in the epididymis of rodents. Here, we examined the expression and localization of Cldn1 and 4 to determine the function of these proteins in the pig epididymis. Cldn1 was highly expressed in the basolateral membrane of epithelial cells in the caput and corpus regions of the epididymis. In the cauda region, however, Cldn1 labeling was significantly decreased in the basolateral membrane of epithelial cells. In contrast, labeling indicated that Cldn4 was expressed in the basolateral membrane in the cauda region of the epididymis and was present at punctate reactive sites in the caput and corpus regions. However, in no region of the epididymis did we detect colocalization of Cldn1 and 4 with labeled ZO-1, the distribution of which is restricted to the tight junctions. Our results indicate that Cldn1 and 4 were region-specifically expressed in the pig epididymis but not present in the tight junctions of epididymal epithelium. In addition, reciprocal regulation in specific regions of the epididymis between Cldn1 and 4 may play an important role in generating an optimal luminal environment for sperm maturation and storage in the pig epididymis.

본 연구는 돈사 내 저장된 돼지분뇨 슬러리에 액상 일라이트와 일라이트 분말을 첨가하여 악취에 미치는 영향을 분석하였다. 공시 돼지는 3원교잡종(Yorkshire×Land race×Duroc) 4개월령 암컷 비육돈 (65~68 kg) 360두를 이용하였다. 시험구 배치는 무처리한 대조구와 돼지분뇨 슬러리에 일라이트 액상 33%와 분말 5%를 첨가한 처리구로 3반복(40두) 120두에 적용하여 완전 임의배치법의 3주간 사양 시험 하였다. 악취 저감제 처리는 슬러리 피트(slurry pit ; 3.75 m×9 m×1.2 m, 20 ton 저장 규모)아래에 저장된 슬러리양을 계산하여 일정한 비율에 맞추고, 매주 액상 일라이트는 분무하거나 또는 일라이트 분말은 뿌려주었다. 악취 측정은 슬러리 피트 위 10 cm 위치에서 암모니아, 황화수소, 메틸 메르캅탄 및 디메틸 설파이드를 측정하였다. 사양기간 동안 암모니아 발생량은 일라이트 처리 형태에 의해 크게 영향을 주었다(p<0.05). 특히 암모니아 발생량은 액상 일라이트 33% 처리구는 30~46%, 일라이트 분말 5% 처리구에서는 30~54% 감소되었다. 황화수소 발생량은 2018년 12월 24일에 측정된 결과를 제외 하고 사양기간 동안 일라이트 처리 형태에 따라 유의적으로 영향을 주었다(p<0.05). 처리구의 황화수 소 발생 감소량은 대조구와 비교할 때, 액상 일라이트 33% 처리구에서는 16~60%, 일라이트 분말 5% 처리구는 7~59%였다. 측정기간 동안 평균 메틸 메르캅탄과 디메틸 설파이드 농도는 모든 시험구에서 발생되지 않았다. 3주간 사양 시험에서 평균 암모니아 농도의 경우, 액상 일라이트 33% 처리구는 38.6%, 일라이트 분말 5% 처리구는 39.7% 감소되었다(p<0.05). 평균 황화수소 농도는 액상 일라이트 33% 처리구와 일라이트 분말 5% 처리구는 비슷한 수준인 42.5% 감소되는 것으로 나타났다(p<0.05). 결론적으로 본 사양시험의 결과 액상 또는 분말 일라이트 처리는 돈사 내 발생되는 암모니아와 황화수소에 대한 유의미한 저감효과를 나타내었다.

Fecal calprotectin is a noninvasive marker of gut inflammation and has been widely utilized in human gastrointestinal diagnostics. This marker, however, has not been extensively utilized in porcine samples. The aim of this study was to optimize a protocol for the extraction of porcine fecal calprotectin, and to the best of our knowledge this is the first study to be conducted in this regard. Freshly collected swine fecal samples were used in this study. We determined the variability of three commercial ELISA assays in the recovery of porcine fecal calprotectin. We further studied the effect of dilution factor and roller shaker homogenization on the yield of calprotectin from swine fecal samples. Calprotectin recovery was significantly different(p<0.05) across the three commercial assays with MBS033848 having a greater recovery compared to DAEF-012 and calprest. Fecal calprotectin yield increased with an increase in dilution factor with maximum recovery at 1:250. Furthermore, homogenization of fecal sample extracts using a roller shaker for tubes for 30 min led to a 30.75% relative increase in calprotectin yield. Further increase in shaking time(at 60 min) led to a reduced calprotectin recovery. Calprotectin recovery ratio was 130.8% and 101.4% at 30 min and 60 min homogenization respectively. In our conclusion, we observed that various factors affect the recovery of porcine fecal calprotectin, and therefore the researcher should double check certain parameters in regard to the type of kit, the dilution factor and homogenization time if reliable and reproducible results are to be obtained. Results of the present study provide useful information on a non-invasive protocol to veterinarians and researchers in examining and monitoring swine gut healthusing the fecal calprotectin.

Cryopreservation is mainly used for preservation of boar sperm. However, this method stresses the sperm by reactive oxygen species (ROS), and the conception rate and the litter size are not more efficient than the liquid preservation of spermatozoa. Therefore, we use chitosan which is a natural product derived antioxidant compound. We used GnHA (chitosan+hyaluronic acid) and GnHG (chitosan hydrogel) as chitosan complexes to cryopreserve boar sperm for improve sperm metabolism and function. Sperm parameter (sperm motility, progressive motility, path velocity, straight-line velocity, curvilinear velocity) is measured by computer-assisted sperm analysis (CASA) using frozen sperm with GnHA or GnHG (0, 0.25, 0.5, 1 mg/mL), respectively. Also, lipid peroxidation analysis using malondialdehyde (MDA) is performed to confirm the antioxidative effect of chitosan in frozen spermatozoa. CASA analysis showed GnHA and GnHG are effective against cryopreserved boar sperm. And antioxidant effect is measured by lipid peroxidation analysis. GnHA and GnHG, which is chitosan complex are effective for boar sperm cryopreservation by antioxidant effect.

Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP × Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (|log2FC| > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

Because of the physiological and immunological similarities between pigs and humans, porcine embryonic stem cells (ESCs) have been identified as important candidates in preliminary studies on human disease. A comparative understanding of pig ESCs with the human is required to achieve these goals. To gain insights into pig stem cells, the transcriptome of pig ES-like cells were compared with pig preimplantation embryos and human/mouse pluripotent stem cells by RNA-seq analysis. As a result, pig stem cells were more similar to late epiblasts of pig preimplantation embryos than early ICM as revealed by transcriptome analysis, suggesting that pig stem cells are in a developmentally primed state. Moreover, the physiological and biological functions of pig ESCs were more similar to those of human PSCs than to those of mouse PSCs, as determined by direct differentiation and GO/KEGG term analysis. Overall, our data indicate that pig ESCs are in a primed pluripotent state resembling human PSCs. Our findings will facilitate both the development of large animal models for human stem cell therapy and the generation of pluripotent stem cells from other domestic animals for agricultural use. This work was supported by the Korea Institute of Planning and Evaluation for Technology in food, agriculture, forestry, and fisheries (IPET) through the Development of High Value-Added Food Technology Program funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA; 118042-03-1-HD020), and partially supported by the grants from the Agenda Program of Rural Development Administration, Republic of Korea (No. PJ01362402)

Because the pig is a valuable candidate for a preclinical model of human cell therapy as well as an important food source, understanding a physiology of pig myogenic progenitors such as skeletal muscle satellite cells and myoblasts is required for cure of muscular diseases and improvement of meat production. For these reasons, we tried to isolate and culture the pig progenitor cells from skeletal muscle. Pig satellite cells, known as muscle stem cells, were isolated from biceps femoris of neonatal pigs by enzymatic digestion method. Muscle satellite cells are quiescent in uninjured muscle. Upon injury, they are activated into proliferating state, known as myoblasts, by growth factors and, in turn, differentiated forward to myocytes and myotubes. To trigger proliferation in vitro, the isolated satellite cells were cultured with epidermal growth factor (EGF) and dexamethasone (BMP4 inhibitor). As a result, the pig satellite cells were efficiently converted into proliferating myoblasts and stably maintained over an extended period. The myoblasts were confirmed by markers of PAX7, MYF5, and MYOD1. Our finding showed that modulating EGF and BMP4 signaling are essential for maintaining muscle stem cells. This culture method could be applied for a production of cultured meat and further basic research of muscle development. This work was supported by the Korea Institute of Planning and Evaluation for Technology in food, agriculture, forestry, and fisheries (IPET) through the Development of High Value-Added Food Technology Program funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA; 118042-03-1-HD020).

This study was conducted to evaluate the effects of insulin and epidermal growth factor (EGF) in a in vitro growth (IVG) medium on oocyte growth, in vitro maturation (IVM) and embryonic development of pig oocytes derived from small antral follicles (SAF) less than 3 mm in diameter. SAF oocytes were cultured for 2 days to induce IVG in alpha-minimal essential medium supplemented with 1 mM dbcAMP and 15% (v/v) fetal bovine serum. After IVG culture, oocyte maturation was induced by culturing IVG oocytes in IVM medium for 44 h. IVM oocytes that extruded the first polar body were selected and induced for parthenogenesis (PA) by applying electric stimulus. SAF oocytes cultured under the insulin treatment showed a significantly increased (P < 0.05) nuclear maturation (73.8%) compared to those cultured with insulin and EGF (59.8%). After PA, the proportions of blastocysts based on the number of metaphase II oocytes were significantly higher (P < 0.05) in oocytes that were cultured for IVG with insulin, EGF, and insulin + EGF (32.4%, 35.2%, and 34.8%, respectively) than in control (22.9%). IVG oocytes treated with insulin showed an increased oocyte diameter (116.3 μm) compared to those treated with insulin and EGF (114.0 μm) (P < 0.05). Intra-oocyte GSH content significantly increased (1.07 pixels/oocyte) by insulin treatment during IVG compared to that of oocytes treated with insulin + EGF (0.78 pixels/oocyte). These results demonstrate that IVG culture of SAF oocytes under insulin or/and EGF treatment supports oocyte maturation and improves embryonic development to the blastocyst stage after PA in pigs.

In our previous studies, the cardiac xenotransplantation from an alpha-1,3-galactosyltransferase knockout pig (GT-MCP-MCP) to cynomolgus monkeys showed a mean survival of 38 days. The objective of this study is to genetically upgrade the GT-MCP-MCP pig, to further enhance membrane cofactor protein (MCP) expression and to express an endothelial specific thrombomodulin (TBM). MCP is a complement regulatory protein and TBM is a coagulation inhibitor. As the dicistronic cassette for wild-type-based MCP and TBM concurrent expressions does not show any increase of MCP, we optimized the MCP codon usage (mMCP) and substituted mMCP for MCP. When the mMCP-TBM cassette was transfected to HeLa cells, we were able to find an increased expression of MCP and endothelial cell-specific TBM expression. The cassette was then transfected into ear-skin fibroblasts isolated from one-month-old #23-4 of a GT-MCP-MCP pig, and the cell populations expressing MCP were obtained by MACS cell sorting. We performed a single cell culture of the selected cells, and obtained clones over expressing 90% MCP. The cells of a clone were used as a donor for nuclear transfer and generated GT-MCP/-MCP/mMCP/TBM pig. The transgenic pig was confirmed to be carrying the cells expressing MCP and functioning as an inhibitor against the cytotoxic effect of normal monkey serum, comparable with donor cells. Thus, we believe that the GT-MCP/-MCP/mMCP/TBM transgenic pig would be potential for the prolongation of xenograft survival in the recipients.