• Journal of the Korean Society of Radiology
• /
• v.5 no.1
• /
• pp.11-18
• /
• 2011

The periosteum contains multipotent cells that can differentiate into osteoblasts and chondrocytes. Cultured periosteum-derived cells (PDCs) have an osteogenic capacity. The purpose of this study was to evaluate the interaction of PDCs with bone graft biomaterial. After cell isolation from the calvarial periosteum of Sprague-Dawley rats, cultured PDCs were placed in critical-sized calvarial defects with beta-tricalcium phosphate (${\beta}$-TCP). All rats were sacrificed 8 weeks after bone graft surgery, and the bone regenerative ability of bone grafting sides was evaluated by plain radiography, micro-computed tomography (CT), and histological examination. PDCs grafted with ${\beta}$-TCP displayed enhanced calcification in the defect site, density of regenerated bone and new bone formation within the defect and its boundaries. Furthermore, these PDCs more efficiently regenerated new bone as compared to grafted ${\beta}$-TCP only. The results suggest that cultured PDCs have the potential to promote osteogenesis in bone defects.

• Korean Journal of Clinical Laboratory Science
• /
• v.51 no.4
• /
• pp.475-483
• /
• 2019

The use of stem cells in cell-based therapy has attracted extensive interest in the field of regenerative medicine, and it has been applied to numerous incurable diseases due to the inherent abilities of self-renewal and differentiation. However, there still exist some severe obstacles, such as requirement of cell expansion before the treatment, and low survival at the treated site. To overcome these disadvantages of stem cells, we used the carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM 6) gene, which functions to increase cell-cell interaction as well as anti-apoptosis. We first confirmed whether CEACAM 6 is expressed in various cell lines at the protein level (including in stem cells), followed by evaluating and selecting the optimal transfection conditions into stem cells. The CEACAM 6 gene was transfected into stem cells to prolong cell survival and preserve from damage by oxidative stress. After confirming the CEACAM 6 expression in transfected stem cells, the cell survival was assessed under oxidative condition by exposing to hydrogen peroxide (H₂O₂) to mimic the chronic environment-induced cellular damage. CEACAM 6 expressing stem cells show increased cell viability compared to the non-CEACAM 6 expressing cells. We propose that the application of the CEACAM 6 gene is a potential option, capable of expanding and enhancing the therapeutic effects of stem cells.

• Journal of Korean Society of Forest Science
• /
• v.79 no.2
• /
• pp.109-114
• /
• 1990

The axillary buds of 15-year-old Tilia amurensis were cultured on Saito and Ide (IS), Murashige and Skoog (MS) media and woody plant medium (WPM) to establish an effective micropropagation method. Five levels of 6-benzylaminopurine (BAP) were tested. On IS medium and WPM addition of 1.0/l BAP enhanced shoot development and shoot elongation, whereas addition of 0.5/l BAP was effective on MS medium. A better results were obtained from WPM with 1.0/l BAP and MS with 0.1/l BAP. Developed shoots were subcultured on each basal media but with 0.2/l BAP, Multiple shoots were almost doubled in a month. Root formation could be enhanced at higher concentration of indole-3-butyric acid (IBA). Better rooting rate (83.3%) was achieved on a half-strength MS medium with 3.0 /l IBA. Regenerated plantlets were successfully transferred to soil. To investigate the clonal variation in shoot development and shoot elongation by axillary bud culturing, seven plus tree clones were tested, Clonal variation in tissue culturability among plus trees was recognized by the Duncan's multiple range test at the 5% level. Kang Won No. 12 showed the best response on WPM with 1.0/l BAP.

• Korean Journal of Weed Science
• /
• v.3 no.2
• /
• pp.129-136
• /
• 1983

To study ecological characteristics of Sagitiaria pygmaea and Sagittaria trifolia occurring in Korea their propagules were collected from 3 locations (Sagittaria pygmaea: Chuncheon, Suweon, Milyang; Sagittaria trifolia: Suweon, Iri, Jeonju) in 1981, cultured and replanted 4 times (May 20, June 5, June 20, July 5) in 1982. Sagitraria pygmaea from Suweon flowered earlier than those from Chuncheon and Milyang in the plants planted on May 20, but this was reversed in another planting dates. Three storied inflorescence was observed newly in Sagittaria pygmaea. Sagittaria pygmaea from Iri and Jeonju had more number of tillers, but less number of tubers per tiller than those from Suweon. Sagittaria rrifolia from Chuncheon flowered earlier than those from Suweon and Milyang. Sagirtaria trifolia from Milyang was narrower in the upper leaf width and less in the number of tubers per plant than those from Chuncheon and Suweon. Each of local collections may be regarded as different ecotype based on the above differences.

• Reproductive and Developmental Biology
• /
• v.31 no.3
• /
• pp.139-143
• /
• 2007

This study was performed to investigate the reproductive characteristics of the cloned Hanwoo bulls produced by SCNT. The semen ejaculated from the cloned bulls (C-38 and C-39) and normal Hanwoo bull was properly measured the volume, the number of sperm, and the viability of frozen-thawed sperm. The sperm activity was analyzed using computer assisted sperm analysis (CASA). To analyze fertilizing ability of the cloned bulls, in vitro fertilization and artificial insemination were performed using the frozen-thawed semen. There were no differences in semen volume, sperm concentration, and the viability of frozen-thawed sperm between cloned bulls and normal bull. The difference was statistically significant in total motility, curvilinear velocity (VCL), straight-line velocity (VSL), and average-path velocity (VAP) of both cloned bulls compared to those of normal Hanwoo bull, respectively (p<0.05). The cleavage and blastocyst development rate were not different between the groups. five cloned cows were artificially inseminated using the frozen-thawed semen of C-38, two of them became pregnant. Two second generation calves (one male and one female) were produced. Based on these results, the cloned Hanwoo bulls showed normal reproductive abilities of semen parameters and sperm activity to their comparators and produced cloned calves, although there are some individual differences on the parameters.

• Journal of Sericultural and Entomological Science
• /
• v.15 no.2
• /
• pp.9-14
• /
• 1973

In order to find out effects of the generative power of silk worm moth which have been brought up in the high temperature accommodation at their pupa stage. For this specific study, 9 different kinds of male silk worms have been selected as specimen. All those specimen were brought up in the normal temperature at their larvae stage and after the pupation period they have been accommodated in the condition of high temperature for a certain length in accordance with the study programme. Afterwards, those mlae specimen were copulated with Suwon jam 103${\times}$104 which were all brought up in normal conditions. This study was carried out to find the copulation function as well as the ratio of unfertilized eggs(male sterility test). Results of study have been revealed as follows: 1) Although some differences were observed, male pura which have been brought up in the condition of high temperature shown the low rate of unfertilized eggs rather than those were brought up in the normal conpition. 2) In this group the eclosion(emergency) has been found to be poor rather, than those specimen brought up in normal conditions. 3) The copulation function of Moran, Daedong, J124 and C108 specimen were found to be poor than those of Suwon jam. 4) Fertility rate of Moran, Daedong, J124 and C108 was found to be around 65%. This figure is rather lower than what we normally expect. 5) Unfertilized egg ratio of Moran, Daedong and C108 were found to be around 20% if they were brought up in the condition of high temperature for one day from the time of pupation: 40% at 2 days, and 70% at 3 days duration. More than 3 days treatment has shown no progress in the unfertilized egg ratio. 6) One day's treatment for the pupa at the later stage has shown the unfertilized egg ratio of about 10%; 20% at 2 day's treatment, 35% at 3 day's treatment, 40-60% at 4 day's treatment, more than 60% at 5 day's treatment, and the 70% of fertilized egg ratio was only observed when the treatment days come to 7 days. It was understood that the unfertilized egg ratio was high at the antepupa stage rather than that of post-pupa stage. 7) According to the result of observation the sperm in copulatory pouch and seminal receptacle out of the normal female silk worm which have been copulated with the male brought up in the condition of high temperature at their moth stage. The reproduction system found in the control zone has been found to be normal and the sperm is amountful and active in motion while the sperm found in the treatment to be limited in amount and slow in motion. The observation was made within 5 hours from the copulation. 8) According to the result of observation of sperm of seminal receptacles of the female silkworm moth, and according to that observation of sperm in the seminal receptacle in female silkworm moth, the amount of sperm and mobility in the female moth brought up in high temperature were poor comparing that were brought up in normal temperature zone. Some of them are even found to be no trace of such. 9) Appearance and mosle of the copulatory organ of the male silkworm moth was found to be no anatomical change. 10) Testis of the later pupa stage which was brought up in the high temperature was found to be almost net developed to anucleate sperm and they are degenerated at stage of between maturation division and sperm abnormal stage. Mean time at control zone, the formation of anucleate sperm was already observed.

• Clinical and Experimental Reproductive Medicine
• /
• v.33 no.1
• /
• pp.25-33
• /
• 2006

Objective: The aim of this study was to investigate whether embryonic stem (ES) cells can be established from isolated blastomeres of mouse embryos. Methods: Blastomeres were separated from mouse (C57Bl/6J) 2- or 4-cell embryos. Isolated blastomeres or whole 4-cell embryos were co-cultured with mitosis-arrested STO feeder cells in DMEM supplemented with recombinant murine leukemia inhibitory factor and ES-qualified fetal bovine serum. After the tentative ES cell lines were maintained from isolated blastomeres or whole embryos, some of them were frozen and the others were sub-cultured continually. Characteristics of tentative ES cell lines as were evaluated for specific genes expressions with immunocytochemistry and RT-PCR. Results: One ES cell line (3.0%) was established from isolated blastomere of 2-cell embryo and one cell line (4.0%) from isolated two blastomeres of 4-cell embryo. And five cell lines (16.7%) were established from whole 4-cell embryos. Both cell lines from isolated blastomere and whole embryo expressed mouse ES cell specific markers such as SSEA-1, Oct-4 and alkaline phosphatase. Marker genes of three germ layers were expressed from embryoid bodies of both cell lines. Conclusion: This study suggests that mouse ES cells could be established from isolated blastomeres, although the efficiency is lower than whole embryos. This animal model could be applied to establishment of autologous human ES cells from biopsied blastomeres of preimplantation embryos in human IVF-ET program.

• Journal of Life Science
• /
• v.17 no.5 s.85
• /
• pp.678-686
• /
• 2007

Human mesenchymal stem cells(hMSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. Therefore, hMSC are attractive candidates for cell and gene therapy. The optimal conditions for hMSC expansion require medium supplemented with fetal bovine serum(FBS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FBS proteins. Previously, we have shown that hADSC can be cultured in human serum(HS) during their isolation and expansion, and that they maintain their proliferative capacity and ability for multilineage differentiation and promote engraftment of peripheral blood-derived CD34 cells mobilized from bone marrow in NOD/SCID mice. In this study we determined whether hADSC grown in HS maintain surface markers expression similar with cells grown in FBS during culture expansion and compared gene expression profile by Affymetrix microarray. Flow cytometry analysis showed that HLA-DR, CD117, CD29 and CD44 expression in HS-cultured hADSC during culture expansion were similar with that in FBS-cultured cells. However, the gene expression profile in HS-cultured hADSC was significantly different from that in FBS-cultured cells. Therefore, these data indicated that HS-cultured hADSC should be used in vivo animal study of hADSC transplantation for direct extrapolation of preclinical data into clinical application.

• Journal of Life Science
• /
• v.21 no.2
• /
• pp.300-308
• /
• 2011

Osteoporosis is a disease involving a decrease in bone mineral density and increased risk of fractures. Osteoblast and osteoclast activities are important for bone formation. The MC3T3-E1 osteoblastic cell line is a well-accepted model of osteogellsis in vitro. Hijikia fusiforme is a kind of edible brown seaweed that grows mainly in the Northwest Pacific region, including the countries of Korea, Japan and China, and it has been widely used as a medicinal and health food in Korea. In this study, by using osteoblasts, the effects of Hijikia fusiforme fractions on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and mineralization of cells were investigated. Hijikia fusiforme were subjected to fractionation by using hexane, methanol, butanol and aqueous. Proliferation of the MC3T3-E1 osteoblastic cells that were treated with Hijikia fusiforme fractions increased by approximately 120%. Regarding effects of Hijikia fusiforme fractions on ALP activity, 1 ${\mu}g$/ml butanol fraction showed the highest activity. The synthesis of collagen increased significantly in response to treatment with Hijikia fusiforme fractions, with the exception of the hexane fraction. Moreover, mineralization in the MC3T3-E1 cells that were treated with 100 ${\mu}g$/ml butanol fraction increased by 281%. Also, when 100 ${\mu}g$/ml aqueous fraction was added, mineralization increased by 240%. These results indicate that Hijikia fusiforme fractions have anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.23 no.1
• /
• pp.70-81
• /
• 2015

Helicases are known to be a proteins that use the chemical energy of NTP binding and hydrolyze to separate the complementary strands of double-stranded nucleic acids to single-stranded nucleic acids. They participate in various cellular metabolism in many organisms. DEAD-box proteins are ATP-dependent RNA helicase that participate in all biochemical steps involving RNA. DEAD-box3 (DDX3) gene is belonging to the DEAD-box family and plays an important role in germ cell development in many organisms including not only vertebrate, but also invertebrate during asexual and sexual reproduction and participates in stem cell differentiation during regeneration. In this study, in order to identify and characterize DDX3 gene in the earthworm, Perionyx excavatus having a powerful regeneration capacity, total RNA was isolated from adult head containing clitellum. Full length of DDX3 gene from P. excavatus, Pe-DDX3, was identified by RT-PCR using the total RNA from head as a template. Pe-DDX3 encoded a putative protein of 607 amino acids and it also has the nine conserved motifs of DEAD-box family, which is characteristic of DEAD-box protein family. It was confirmed that Pe-DDX3 has the nine conserved motifs by the comparison of entire amino acids sequence of Pe-DDX3 with other species of different taxa. Phylogenetic analysis revealed that Pe-DDX3 belongs to a DDX3 (PL10) subgroup of DEAD-box protein family. And it displayed a high homology with PL10a, b from P. dumerilii.