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This study aimed to determine the cellular characteristics and behaviors of human bone marrow stromal cells (hBMSCs) expanded in media in a hypoxic or normoxic condition and with or without fibroblast growth factor-2 (FGF-2) treatment. hBMSCs isolated from the vertebral body and expanded in these four groups were evaluated for cellular proliferation/migration, colony-forming units, cell-surface characterization, in vitro differentiation, in vivo transplantation, and gene expression. Culturing hBMSCs using a particular environmental factor (hypoxia) and with the addition of FGF-2 increased the cellular proliferation rate while enhancing the regenerative potential, modulated the multipotency-related processes (enhanced chondrogenesis-related processes/osteogenesis, but reduced adipogenesis), and increased cellular migration and collagen formation. The gene expression levels in the experimental samples showed activation of the hypoxia-inducible factor-1 pathway and glycolysis in the hypoxic condition, with this not being affected by the addition of FGF-2. The concurrent application of hypoxia and FGF-2 could provide a favorable condition for culturing hBMSCs to be used in clinical applications associated with bone tissue engineering, due to the enhancement of cellular proliferation and regenerative potential. Keywords: Bone marrow stromal cells, Hypoxia, Fibroblast growth factor, Tissue regeneration, Microenvironment interactions

Centrifugation based on density gradients is a general methodology for isolating human bone marrow (hBM)-derived mesenchymal stem cells (hBMSCs). The mononuclear cell (MNC) layer can be obtained using a density gradient solution in the conventional protocol, but it is not suitable for direct transplantation due to the possible toxicity of this solution. The results obtained are also influenced by the skill level when applying the technique, which involves time-consuming processes. We have developed a novel protocol for isolating hBMSCs using hyaluronic acid (HA), which is the most widely used injectable biomaterial in clinical settings and a major component of the extracellular matrix. Laying hBM over the HA and then applying centrifugation yielded three separate layers, with the HA layer, including MNCs being the most superficial one. Increasing the volume of HA and/or its crosslinking rate enhanced the yield of MNCs from hBM, and the cell yield was also significantly higher for a lower centrifugal acceleration (530 g) than for a higher one (1500 g). Isolated hBMSCs by HA exhibited similar biological characteristics such as in terms of their proliferation rate, fibroblast-like morphology, cell-cycle status, immunophenotype, and multipotency. The use of either type of hBMSC confirmed the regenerative potential of bone and bone marrow-like tissue in ectopic transplantation models. This is the first report of a novel protocol for isolating hBMSCs that utilize HA. We suggest that this novel isolation technique can be used for the direct application of autogenous MSCs with advantages of being less time-consuming and involving steps that are easier to perform.

Many studies have reported the frequency and distribution of haplogroups among various cattle breeds for verification of their origins and genetic diversity. In this study, 318 complete sequences of the mtDNA control region from four Korean cattle breeds were used for haplogroup classification. 71 polymorphic sites and 66 haplotypes were found in these sequences. Consistent with the genetic patterns in previous reports, four haplogroups (T1, T2, T3, and T4) were identified in Korean cattle breeds. In addition, T1 a, T3a, and T3b sub-haplogroups were classified. In the phylogenetic tree, each haplogroup formed an independent cluster. The frequencies of T3, T4, T1 (containing T1a), and T2 were 66%, 16%, 10%, and 8%, respectively. Especially, the T1 haplogroup contained only one haplotype and a sample. All four haplogroups were found in Chikso, Jeju black and Hanwoo. However, only the T3 and T4 haplogroups appeared in Heugu, and most Chikso populations showed a partial of four haplogroups. These results will be useful for stable conservation and efficient management of Korean cattle breeds.

This study aimed to identify allele variability and determine the frequencies of microsatellite (MS) markers specific to bovine sex chromosomes in Korean native cattle (KNC; Chikso, Hanwoo and Jeju Black). The three KNC breeds were characterized using 4 MS markers (INRA30, TGLA325, UMN0905 and UMN0929), and allelic richness, allele frequency, heterozygosity and polymorphism information content (PIC) per locus were calculated. The mean observed heterozygosity and PIC values for Chikso, Jeju Black and Hanwoo were 0.541 and 0.542, 0.406 and 0.377, and 0.61 and 0.6, respectively. The heterozygosity and the PIC values of Jeju Black breed were smaller than those of Chikso and Hanwoo. Thus, these genetic parameters were used for the characterization of the breeds and for determining their genetic relationships. The findings of this study might be useful for the conservation, management, and utilization of KNC breeds as genetic resources.

The goals of this study were to identify sequence variations in the mitochondrial cytochrome b (mtDNA cyt b) gene in White Hanwoo (Wh) and the genetic relationship between the Wh and other breeds. When whole sequences of the mtDNA cyt b gene in 14 Wh cattle were determined, a silent mutation and two haplotypes were detected in the Wh cattle. The major haplotype, H1, was found in 13 of 14 individuals in the Wh cattle. Haplotype diversity and nucleotide diversity were 0.143 and 0.00013, respectively. Compared to previous reports, these levels of genetic diversity are lower than other Korean and Chinese breeds. To identify the genetic relati onship among Korean, Chinese, Japanese, and European cattle breeds, the neighbor-joining (NJ) tree was constructed based on Dxy genetic distances. Two distinct groups were identified and classified as A and B. Wh was found in the A group, which consisted of Bos taurus breeds. From calculating the Dxy genetic distances, Wh was found to be genetically more closely related to two breeds, Heugu (0.00018) and Yanbian (0.00021), than to other breeds. In conclusion, Wh is genetically related to Chikso, Heugu, and Yanbian breeds based on maternal inheritance. The results of this study will be useful for efficient management and sustainable utilization of Wh.

Four Korean native cattle (KNC) breeds-Hanwoo, Chikso, Heugu, and Jeju black-are entered in the Domestic Animal Diversity Information System of the United Nations Food and Agriculture Organization (FAO). The objective of this study was to assess the genetic diversity, phylogenetic relationships and population structure of these KNC breeds (n = 120) and exotic breeds (Holstein and Charolais, n = 56). Thirty microsatellite loci recommended by the International Society for Animal Genetics/FAO were genotyped. These genotypes were used to determine the allele frequencies, allelic richness, heterozygosity and polymorphism information content per locus and breed. Genetic diversity was lower in Heugu and Jeju black breeds. Phylogenetic analysis, Factorial Correspondence Analysis and genetic clustering grouped each breed in its own cluster, which supported the genetic uniqueness of the KNC breeds. These results will be useful for conservation and management of KNC breeds as animal genetic resources.

Domestic goats (Capra hircus) are classified into six different mitochondrial DNA (mtDNA) haplogroups (A, B, C, D, F, and G), of which haplogroup A shows worldwide distribution. However, there is currently no key for genetic and geographic classification of haplogroup A. In this study, we determined the complete sequences of the mtDNA control region from Korean native goat breeds. In addition, we compared these sequences with those of other domestic goat breeds/populations globally. A distinct variation site was found at position 717. Of two types (C and T) from this variation, C type sequences were found only in haplogroup A, and were specific to Eastern Asia. On the other hand, T type sequences were distributed in all haplogroups. Phylogenetic analysis divided the two types according to different phylogenetic locations. These results suggest that a variation in position 717 is a key for geographical classification of mtDNA haplogroup A. Moreover, these distribution patterns might provide for identification of the maternal origin of goat genetic resources, including Korean native goat breeds.

The vent sexing and the auto-sexing by using sex-linked traits are general sexing methods of day-old chicks. Currently, the feather sexing which is based on the differences in the feather characteristics at hatching is the representative sexing method of chicken, because the late-feathering is sex-linked trait. The feather sexing can be used if the breed has dominant feathering gene (K) in maternal and recessive gene (k + ) in paternal. Therefore it is necessary to identify the associa- tion of feathering genes and quantitative traits in chickens. In this study, we investigated the influence of the rate of feathering on productive traits in Korean Native Chicken. In results, there was no significant difference between early-feathering chickens and late-feathering chickens in reproductive performance such as fertility and hatchability. Livability, body weights, egg production, egg weight and egg quality also did not significantly differ between early- and late-feathering chickens. Age at first egg was the only trait of those tested in which significant difference was observed. The early-feathering chickens laid eggs 3 days earlier than late-feathering chicken. As a result, there is no influence of feathering phenotypes on productive performance in Korean Native Chickens. Consequentially, establishing the feather sexing strain is available using the Korean Native Chicken breed without considering of the effect of feathering genes on productive traits.

Microsatellite markers have been a useful genetic tool in determining diversity, relationships and individual discrimination studies of livestock. The level of genetic diversity, relationships among two Korean indigenous chicken brand populations (Woorimatdag: WR, Hanhyup3: HH) as well as two pure populations (White Leghorn: WL, Rhode Island Red: RIR) were analyzed, based on 26 MS markers. A total of 191 distinct alleles were observed across the four chicken populations, and 47 (24.6%) of these alleles were unique to only one population. The mean HExp and PIC were estimated as 0.667 and 0.630. Nei's DA genetic distance and factorial correspondence analysis (FCA) showed that the four populations represented four distinct groups. However, the genetic distance between each Korean indigenous chicken brand (WR, HH) and the pure population (WL, RIR) were threefold that among the WR and HH. For the STRUCTURE analyses, the most appropriate number of clusters for modeling the data was determined to be three. The expected probabilities of identity among genotypes of random individuals (PI) were calculated as 1.17×10 49 (All 26 markers) and 1.14×10 15 , 7.33×10 20 (9, 12 with the highest PI value, respectively). The results indicated that the brand chicken breed traceability system employing the own highest PI value 9 to 12 markers, and might be applicable to individual identification of Korean indigenous chicken brand. (Key words : Microsatellite, Genetic relationship, Korean indigenous chicken brand, Identification)

In order to analyze the genetic diversity and phylogenetic status of the Korean Chikso breed, we determined sequences of mtDNA cytochrome b (cyt b) gene and performed phylogenetic analysis using 239 individuals from 5 Chikso populations. Five non-synonymous mutations of a total of 15 polymorphic sites were identified among 239 cyt b coding sequences. Thirteen haplotypes were defined, and haplotype diversity was 0.4709 ranging from 0.2577 to 0.6114. Thirty-five haplotypes (C1-C35) were classified among 9 Asia and 3 European breeds. C2 was a major haplotype that contained 206 sequences (64.6%) from all breeds used. C3-C13 haplotypes were Chikso-specific haplotypes. C1 and C2 haplotypes contained 80.5% of cyt b sequences of Hanwoo, Yanbian, Zaosheng and JB breeds. In phylogenetic analyses, the Chikso breed was contained into B. taurus lineage and was genetically more closely related to two Chinese breeds than to Korean brown cattle, Hanwoo. These results suggest that Chikso and Hanwoo have a genetic difference based on the mtDNA cyt b gene as well as their coat color, sufficient for classification as a separate breed.

The purpose of this study was to identify genetic polymorphisms of the mitochondrial cytochrome b (mtDNA cyt b) gene in Korean black (KB) cattle breed and to analyze the genetic relationship between the KB and other breeds. We determined the complete sequence of the mtDNA cyt b gene in 38 KB cattle. We also analyzed their genetic diversity, and phylogenetic analysis was performed by comparison with Korean cattle (KC, called Hanwoo) and breeds from China and Japan. A nucleotide substitution was detected in the KB cattle, and two haplotypes were defined. In the neighbor-joining (NJ) tree, the haplotypes of KB were located in Bos taurus lineage with those of KC, Japanese black (JB), Yanbian and Zaosheng breeds. However, the haplotypes of Chinese breeds, excluding Yanbian and Zaosheng, were separated into B. taurus and B. indicus lineages. In the NJ tree of breeds based on Dxy genetic distances, Chinese breeds mixed with B. taurus and B. indicus lineages were located between B. indicus and B. taurus lineages. KB was contained within B. taurus lineage and was determined to be genetically more closely related to two Chinese (Yanbian and Zaosheng) breeds than to KC and JB. The haplotype distribution and the results of the phylogenetic analysis suggest that KB and KC have genetic differences in their mtDNA cyt b gene sequences.

Department of Animal Science, Chonnam National UniversityABSTRACT The objective of this study was to compare the growth performance between Korean native layer chickens and imported layer chickens at early rearing stage. Total number of chicks analyzed in this study was 276 and feeding period was conducted from July 24, 2012 for 10 weeks. Five strains including 2 Korean native strains: A=Korean Native Black (Chung-cheongbuk-do) and B=Korean Native Yellowish Brown (Gyeongsangbuk-do) and 3 imported layer strains: C=White Leghorn (Gyeongsangnam-do), D=White Leghorn (Seoul), and E=Ameraucanas (Gyeongsangbuk-do) were used to analyze the following traits such as fertility, hatchability, body weight at a different growing stage, average body weight gain, and feed conversion ratio. The fertilities and hatchabilities of strains were 93.88% and 95.65% in strain A, 81.75% and 86.24% in strain B, 82.25% and 88.15% in strain C, 79.25% and 90.85% in strain D, and 71.50% and 88.11% in strain E, respectively. A viability was excellent in strains A and E to be more than 98% and was low in strain D to be 86.67% at a whole week. The strain A had greater body weight during growing stages (p

본 연구는 30개의 MS 마커를 이용하여 한국재래염소 3개 집단(당진, 장수, 통영)과 1개 농가집단을 대상으로 집단 내 및 집단간의 유전적 다양성, 계통유전학적 유연관계 분석 및 한국재래염소 3개 집단간의 유전적 균일성을 검증하여 우리 고유유전자원으로서의 가치를 구명하고자 실시하였다. 대립유전자형 분석 결과, 총...

The goal of this study was to verify the phylogenetic status of the Korean native goats (KNG). We determined the complete sequence of the mitochondrial cytochrome b gene in 48 goats among four populations. We also analyzed genetic variability within goats, and a phylogenetic analysis was performed by comparison with other country's goats. Three nucleotide substitutions were detected, and two of these were missense mutations that occurred due to a substitution of amino acid. Four haplotypes were defined from KNG. Three of these haplotypes were only found in the Chinese goat. However, the other haplotype was KNG-specific. In the phylogenetic analysis, four clades (A~D) were classified among domestic goats, and the KNG was classified into clade 1 that estimated as lineage A based on the D-loop sequence. Each haplotype from the KNG was clustered closely with that of the Chinese goat. The results of haplotype distribution and phylogenetic location suggest that strong gene flow occurred from China to the Korean Peninsula. (Key words : Korean native goat, Phylogenetic analysis, Cytochrome b, Haplotype, Gene flow)

Genetic resource conservation of indigenous species is important to conserve terrestrial, aquatic and avian species throughout the world. The native Korean Oge (KO) chickens are important and protected indigenous avian species in Korea. This KO breed is very popular in Korea because of its external appearances of blackish color. The characterization of morphology and growth performances of KO were performed since 1980s, however the information is still not sufficient for breed selection. In this study, we compared various growth performances including body growth, tibia length, shank length, chest width, chest grith and length of tail feather between male and female chickens of KO and widely known White Leghorn (WL) breeds at 1 week, 2 weeks, 5 weeks, 10 weeks, and 24 weeks. We observed differences on various growth performances at different aged groups between KO and WL chickens. This study may help for the selection of chicken breeds based on age, body growth and meat production.

Korean native goats, which are characterized by black coat color, have existed on the Korean peninsula for a long time. Until now, there has been no comprehensive investigation concerning their genetic diversity, phylogenetic analysis or origin. In this study, we investigated the genetic diversity and verified phylogenetic status of the Korean native goat using the 453-bp fragment of the hypervariable fragment I (HVI) of mitochondrial DNA (mtDNA) D-loop region from 60 individuals among 5 populations. The Korean native goat showed less haplotype diversity when compared with goats from other countries. In addition, 6 haplotypes that had not been previously reported were verified in this study. In phylogenetic analyses with other country"fs goats, 10 haplotypes from Korean native goats were classified into mtDNA lineage A. Moreover, in a phylogenetic tree for goats which contained mtDNA lineage A, 8 of 10 haplotypes could be included in a subgroup with goats from Vietnam and an area of China. However, none of the remaining haplotypes belonged to a major group of Korean native goats and were located on different independent positions. These results suggest that almost Korean native goats aligned more closely to China and Vietnam breeds in mtDNA lineage A and there was no gene flow from other mtDNA lineages. Our results will contribute to conservation strategies and genetic breeding of Korean native goats.

Two sets of data on carcass traits and beef cut parameters were used to investigate the relationships between carcass and beef cut measurements, which can be used to make predictions of retail cut percentages. One set had a total of 1,141 measurements of Hanwoo cattle of three different sex origins, which were slaughtered in an abattoir located at the National Institute of Animal Science, RDA, Korea from 1996 to 2008. To develop prediction models for retail cut percentage with higher accuracies than the current model, another set consisting of a total of 13,389 records of carcass and beef cut traits were collected from 30 abattoirs and butcheries in Korea from 2008 to 2009. Bulls yielded heavier and leaner carcasses than steers. High correlation coefficients were estimated between amount of body fat and percent retail cut (-0.82) as well as between back fat thickness (BF) and percent retail cut (-0.62). The amount of retail cut, however, was highly correlated with body weight before slaughter (BW, 0.95) or with cold carcass weight (CWT, 0.94). Relationships between percent retail cut and measurable beef yield traits, BF, loin eye area (LEA) or CWT varied by sex class, which must be considered for development of a prediction model with high accuracy. Models of data for all breeds and sexes fit the effects of breed, sex, and interaction of abattoir by butchers, whereas models of data for each breed and sex fit the effect of interaction of abattoir by butcher only. Due to possible future changes in back fat control, we performed a log transformation of BF. Our new models fit better than the currently used model.

Lead, copper, and cadmium were adsorbed onto aged calcium alginate beads containing cell-suspension from the waste of beer fermentation broth. Beads prepared by adding 0.6% (w/v) sodium alginate into the cell suspension from the waste of beer fermentation broth and making the cell suspension drop into the 1% (w/v) calcium alginate solution were stored in the 1% (w/v) calcium chloride solution for 1 year. The specific metal uptake of the aged cell-suspension immobilized bead was 312 mg Pb2+, 158 mg Cu2+, and 112 mg Cd2+/g bead dry weight at pH 7.5 of the metal solution, respectively. The relation between the specific metal uptake by the aged cell-suspension immobilized beads and the equilibrium metal concentration was nonlinearly regressed and well described by the Freundlich isotherm. The specific cadmium uptake capacity of aged cell-suspension immobilized beads was between the specific cadmium uptake capacities of commercial beads Duolite GT-73 and Amberite IRA-400 and higher than those of the fresh Saccharomyces cerevisiae ATCC 834 and Saccharomyces cerevisiae ATCC 24858 immobilized beads.

In order to investigate the effect of feeding browses on the incidence of abortion in pregnant Korean native female goats, 20 pregnant does(34 month) were used as experimental animal and they grouped 4 treatments with 5 heads according to experimental diets. Each group was fed pine browse, pine browse silage, fermented pine browse, or oak browse silage. Browses intake a day was 0.36kg for oak browse silage, 0.28kg for pine browse, 0.24kg for pine browse silage, and 0.14kg for fermented pine browse. Salmonella and Fungi were not found in pine browses but they were found in oak browse silage in the amount of 7.93 and 11.1 cfu/g, respectively. E. Coliwas found 11.67cfu/g in oak browse silage. The incidence of abortion was 60% for fermented pine browse, 40% for pine browse silage, and 20% for pine browse feeding. Abortion did not occur by feeding oak browse silage. Progesterone concentration was similar each other regardless of normal delivery or abortion at delivery day but the concentration of estradiol was higher for normal delivery, concentration of cortisol was decreased until the delivery day in normal but increased in abortion does. The results were suggested.

Data were collected from 1996 through 2000 on Korean Native Goats by the National Livestock Research Institute of Korea were used to estimate genetic parameters for birth, 3 month, and 6 month body weights. Estimates were obtained with MTDFREML. Model included animal and maternal genetic and residual effects. The model included sex, birth year-season, and feeding type as fixed factors. Average body weights and standard deviation were 1.780.32 at birth of age, 7.992.66 at 3 month of age, and 12.083.20 kg at 6 month of age, respectively. Average body measurements were 36.46cm for withers height, 38.06cm for body length, and 45.56cm for heart girth at 3 month of age, and were 40.27cm for withers height, 42.01cm for body length, and 51.07cm for heart girth at 6 month of age, respectively. Estimates of heritability were 0.66 for birth weight, 0.34 for 3 month body weight, and 0.27 for 6 month body weight, respectively. Maternal effects would be important for birth and 3 month body weights and may not be needed in a model for 6 month body weight.

The effective management of endangered animal genetic resources is one of the most important concerns of modern breeding. Evaluation of genetic diversity and relationship of local breeds is an important factor towards the identification of unique and valuable genetic resources. This study aimed to analyze the genetic diversity and population structure of six Korean native chicken breeds (n = 300), which were compared with three imported breeds in Korea (n = 150). For the analysis of genetic diversity, 30 microsatellite markers from FAO/ISAG recommended diversity panel or previously reported microsatellite markers were used. The number of alleles ranged from 2 to 15 per locus, with a mean of 8.13. The average observed heterozygosity within native breeds varied between 0.46 and 0.59. The overall heterozygote deficiency (F-IT) in native chicken was 0.234 +/- 0.025. Over 30.7% of F-IT was contributed by within-population deficiency (F-IS). Bayesian clustering analysis, using the STRUCTURE software suggested 9 clusters. This study may provide the background for future studies to identify the genetic uniqueness of the Korean native chicken breeds.

The importance of genetic resource preservation has been highlighted in the literature as a means of maintaining genetic diversity. Among the various methods of preserving such resources, semen cryopreservation can be advantageous because it reduces the time of restoring genetic resources and is less technique-dependent. The Korean Oge (KO) chicken is a Natural Monument and is recognized as an important genetic resource in Korea. However, successful cryopreservation methods for KO chickens have yet to be reported. Therefore, we completed cryopreservation methods in KO chickens using N-methylacetamide (MA) as a cryoprotectant. Also we performed additional experiments to identify whether fertility and hatchability are affected by long-term storage. Finally, we examined sperm viability in the cryopreserved semen. Our results suggest that the cryopreservation method using MA can be applied to KO chickens regardless of storage period and could be a useful tool for the preservation the endangered avian species.