• Food Science of Animal Resources
• /
• v.40 no.5
• /
• pp.852-859
• /
• 2020

The muscle stem cells of domestic animals are of interest to researchers in the food and biotechnology industries for the production of cultured meat. For producing cultured meat, it is crucial for muscle stem cells to be efficiently isolated and stably maintained in vitro on a large scale. In the present study, we aimed to optimize the method for the enrichment of pig muscle stem cells using a magnetic-activated cell sorting (MACS) system. Pig muscle stem cells were collected from the biceps femoris muscles of 14 d-old pigs of three breeds [Landrace×Yorkshire×Duroc (LYD), Berkshire, and Korean native pigs] and cultured in skeletal muscle growth medium-2 (SkGM-2) supplemented with epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580). Approximately 30% of total cultured cells were nonmyogenic cells in the absence of purification in our system, as determined by immunostaining for cluster of differentiation 56 (CD56) and CD29, which are known markers of muscle stem cells. Interestingly, following MACS isolation using the CD29 antibody, the proportion of CD56⁺/CD29⁺ muscle stem cells was significantly increased (91.5±2.40%), and the proportion of CD56 single-positive nonmyogenic cells was dramatically decreased. Furthermore, we verified that this method worked well for purifying muscle stem cells in the three pig breeds. Accordingly, we found that CD29 is a valuable candidate among the various marker genes for the isolation of pig muscle stem cells and developed a simple sorting method based on a single antibody to this protein.

• Journal of Animal Science and Technology
• /
• v.49 no.5
• /
• pp.677-688
• /
• 2007

돼지를 포함한 대부분의 동물은 일정한 발정주기를 가지고 일정한 시기에 배란을 하는 자연배란동물이지만, 토끼, 고양이, 밍크 등의 암놈은 교미자극에 의해 배란이 일어나는 유기배란동물이다. 또한 1년에 한 번만 발정하는 단발정동물과 1년에 수차례 발정하는 다발정동물이 있다. 이 중에서 모돈은 1년에 수차례 발정하는 다발정 동물로서 발정기에 들면 비발정기와는 다른 행동을 나타낸다(Diehl 등, 2001). 양돈가의 수익을 최대화하기 위해서는 비생산일수를 최소로 줄여야 한다. 모돈의 비생산일수를 줄일 수 있는 한 가지 방법은 성공적으로 교배를 시키는 것이다. 이처럼 성공적으로 교배를 시키기 위해서는 수정적기를 정확히 예측해야 한다. 만약 수정적기를 정확히 판단하지 못하여 수태가 되지 않으면, 비생산일수가 늘어나 손실을 입게 된다. 따라서 수정적기를 정확히 판단하는 것은 모돈의 성공적인 인공수정에 있어서 중요한 요소이다. 수정적기는 배란이 일어나기 전 10시간에서 12시간 사이이며, 발정이 시작되는 시점을 기준으로 하였을 때 경산돈의 경우 26시간에서 34시간 사이이고 미경산돈의 경우는 18시간에서 26시간 사이이다(Evans 등, 2001). 현재 하루에 두 번 모돈의 발정을 확인하는 것이 일반화되어 있으며, 이 때 웅돈을 접촉시키거나 육안관찰을 통하여 발정 유무를 판단한다. 이러한 방법에는 숙련된 기술과 풍부한 경험이 요구될 뿐만 아니라 총 소요노동력의 30% 정도가 요구된다(Perez 등, 1986). 하루에 두 번밖에 발정을 감지하지 않기 때문에 발정이 언제 시작되었는지를 정확히 알 수 없으며, 또한 발정의 대부분이 새벽에 시작되므로 수정적기를 정확히 판단하기란 매우 어렵다. 만약 발정을 감지했더라도 적기에 인공수정을 하지 못한다면, 수태율이 낮아지므로 경제적 손실이 초래된다. 현재 이러한 문제점 때문에 2회에서 3회에 걸쳐 인공수정을 하고 있으나 이에 따른 소요비용과 소요노동력 등은 양돈가의 부담을 가중시키는 요인이 되고 있다. 돼지는 발정기가 되면 비발정기에 나타내지 않던 외음부의 냄새를 맡는 행동, 귀를 세우는 행동 및 승가허용 행동 등을 나타낸다(Diehl 등, 2001). 또한 돼지는 비발정기에 비하여 발정기에 더 많은 활동량을 나타낸다(Altman, 1941; Erez and Hartsock, 1990). Freson 등(1998)은 스톨에서 개별적으로 사육되고 있는 모돈의 활동량을 적외선센서를 이용하여 측정함으로써 발정을 86%까지 감지하였다고 보고하였다. 그러나 이 연구는 단지 모돈의 발정을 감지하였을 뿐 번식관리에 있어서 가장 중요한 수정적기의 판단 기준을 제시하지 못하였다. 따라서, 본 연구는 스톨에서 사육되는 모돈의 활동량을 측정함으로써 발정시작시각을 감지하고 이를 기준으로 인공수정적기를 예측할 수 있는 인공수정적기 예측 장치를 개발한 후 이의 성능을 농장실증실험을 통하여 시험하고자 수행되었다.

• Journal of Life Science
• /
• v.26 no.11
• /
• pp.1232-1236
• /
• 2016

Mitochondrial calcium uptake 1 (MICU1) including two canonical EF hands, located in the mitochondrial inner membrane, is known to play a crucial role in the calcium uptake in mitochondria. Mitochondrial calcium uptake in muscular cells is related to post mortem shortening by calcium release from muscles. Therefore, the porcine MICU1 gene has been estimated as a genetic candidate for meat quality traits. In this study, variations on the exonic regions of the porcine MICU1 gene were investigated by sequencing cDNAs and tested for their association with meat quality traits. A total of 667 Berkshire heads (347 sows and 320 castrated boars) were used for this association test. Three SNPs were detected on the 3' untranslated region (UTR) of the porcine MICU1 gene. SNP1 (c.*136G>A) was associated with drip loss (p=0.017) and intramuscular fat content (p=0.039). In addition, SNP2 (c.*222G>A) and SNP3 (c.*485G>A) were associated with drip loss (p=0.018) and intramuscular fat content (p<0.001), respectively. In conclusion, it was verified that three variations on the 3' UTR of the porcine MICU1 gene were significantly associated with meat quality traits. It was also suggested that molecular biological analyses are needed to validate the function of variations on the 3 UTR of the porcine MICU1 gene.

• Journal of Animal Science and Technology
• /
• v.44 no.2
• /
• pp.207-212
• /
• 2002

This study was conducted to investigate the genotypes and frequencies of Melanocortin 1 Receptor(MC1R) genes in pigs which plays a central role in regulation of eumelanin (black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Four different breeds of pigs(20 Landrace, 20 Yorkshire, 20 Duroc, and 93 Jeju native black pigs) were used and PCR-RFLP analysis of MC1R gene was also carried out. Two regions of MC1R genes (428bp and 405bp) were amplified using two specific primers (MERL1-EPIG2, EPIG1-EPIG3), respectively and MC1R allele were determined using 2 restriction enzymes (BspHⅠ, AccⅡ). The results of this experiment indicated that MC1R allelic type in Landrace, Large Yorkshire and Duroc were MC1R *2 (Ep), MC1R *2 (Ep), MC1R *4 (e), respectively. However, various allelic types of MC1R genes were detected in Jeju native black pigs. MC1R allelic type of Jeju black pigs was MC1R*2 type as in Meishan and Large black breeds or MC1R*3 type as in Hampshire and Berkshire breeds and the gene frequencies of ED1 and ED2 were 0.554 and 0.446 in average.

• Food Science of Animal Resources
• /
• v.28 no.2
• /
• pp.171-180
• /
• 2008

This study was conducted to investigate the meat quality characteristics of pork loin in 3 crossbred groups (30 pigs/group). The crossbreeds were LY female${\times}$D male, Y female${\times}$B male and L female${\times}$B male (L: Landrace, Y: Yorkshire, B: Berkshire, D: Duroc). Pork loins were collected from animals ($110{\times}120kg$ body weight) slaughtered in a commercial slaughter house and stored at $-3^{\circ}C$. Samples were analyzed for general composition, physico-chemical properties, meat and fat color, texture characteristics, sensory evaluation and fatty acid composition at 24 hrs postmortem, and TBARS (Thiobarbituric Acid Reactive Substances) values were determined at various storage times. The LYD pigs showed a higher back fat thickness than the YB and LB pigs. There were no significant differences in intramuscular fat, intermuscular fat, subcutaneous fat and springiness among the 3 crossbreeds. In addition, there were no significant differences in general composition, pH, drip loss, cooking loss and cooked sensory evaluation among the 3 crossbreeds (p>0.05). The shear force value of LB pork was significantly lower than other crossbreeds (p<0.05). Total heme pigment was significantly higher in YB pork compared to LYD and LB pork (p<0.05). The TBARS value of YB pork was significantly lower than that of LYD pork, except after the I day of storage (p>0.05). TBARS values increased significantly during storage for all three crossbreeds. The lightness (CIE L) of YB pork was significantly lower, while the redness (CIE a) was significantly higher in YB pork relative to the other crossbreeds. With regard to textural properties, the hardness and adhesiveness values were significantly lower in YB pork compared to LB pork. In fresh meat, the color of YB pork was significantly higher than the other breeds (p<0.05). The content of myristic acid (C14:0) was significantly lower, while the content of arachidonic acid (C20:4) was significantly higher in LB pork (p<0.05). These results show that the YB breed has the highest quality meat of the three crossbreeds.

• Journal of Animal Science and Technology
• /
• v.54 no.1
• /
• pp.1-8
• /
• 2012

This study aimed to investigate the single nucleotide polymorphisms (SNPs) of the porcine MC4R gene and validate the effect of the MC4R genotype for marker assisted selection (MAS). Six amplicons were produced to analyze the entire base sequences of the porcine MC4R gene and six SNPs were detected (c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, c.892A>G-Asp298Asn, and c.*430A>T). Linkage disequilibrium (LD) of the six SNPs was analyzed by performing haploid analysis. There was a perfect linkage disequilibrium in c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, and c.*430A>T. Only the c.892A>G (Asp298Asn) SNP showed a very low LD with an $r^2$ value of 0.028 and the D' value of 0.348. As a result, the two SNPs-c.707A>G (Arg236His) and c.892A>G (Asp298Asn)-were selected to extract the genotype frequencies from the 5 pig breeds by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotype analysis method. The SNP frequency of c.707A>G (Arg236His) indicated the presence of the A (His) allele only in Yorkshire, while the G allele was fixed in the KNP, Landrace, Berkshire, and Duroc. Association analysis was carried out in 484 pigs with the c.707A>G (Arg236His) SNP and the meat quality traits of four different pig cross populations: a significant association was noted in crude fat, sirloin moisture, meat color, and the degree of red and yellow coloration. The frequency of the c.892A>G(Asp298Asn) SNP genotype varied among the breeds; while Duroc showed the highest frequency of the A (Asn) allele, KNP showed the highest frequency of the G (Asp) allele. Association analysis was carried out in 1126 pigs with the c.892A>G (Asp298Asn) SNP and the meat quality traits of four pig populations: a highly significant linkage was noted in the back-fat thickness (P<0.002). It was found that the back-fat thickness was higher in individuals with the AA genotype than in those with the AG or GG genotype. Thus, in this study, we verified that the c.892A>G (Asp298Asn) SNP in the pig MC4R gene has a sufficient effect as a gene marker for MAS in Korean pork industry.