• Title/Summary/Keyword: Cell formation

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Differentiation of Osteoblast Progenitor Cells from Human Umbilical Cord Blood (제대혈액에서 골조직 특이세포로의 분화)

  • Hong, Seung-Jin;Lee, Eun-A;Chae, Gue-Tae;Han, Hoon
    • IMMUNE NETWORK
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    • v.2 no.3
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    • pp.166-174
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    • 2002
  • Background: Human umbilical cord bloods, which could be taken during the delivery are utilized as a source of hematopoietic stem cells. Also in cord blood, there are several kinds of stem cells such as endothelial and mesenchymal stem cells. Methods: We isolated the mesenchymal stem cells from human umbilical cord bloods and confirmed the differentiation of these cells into osteoblast progenitor cells. The mesenchymal stem cells derived from umbilical cord blood have the ability to differentiate into specific tissue cells, which is one of characteristics of stem cells. These cells were originated from the multipolar shaped cells out of adherent cells of the umbilical cord blood mononuclear cell culture. Results: The mesenchymal stem cells expressed cell surface antigen CD13, CD90, CD102, CD105, ${\alpha}$-smooth muscle actin and cytoplasmic antigen vimentine. Having cultrued these cells in bone formation media, we observed the formation of extracellular matrix and the expression of alkaline phosphatase and of mRNA of cbfa-1, ostoecalcin and type I collagen. Conclusion: From these results we concluded that the cells isolated from the umbilical cord blood were mesenchymal stem cells, which we could differentiate into osteoblast when cultured in bone formation media. In short, it is suggested that these cells could be used as a new source of stem cells, which has the probability to alternate the embryonic stem cells.

Supplement of tauroursodeoxycholic acid in vitrification solution improves the development of mouse embryos

  • Lin, Tao;Lee, Jae-Eun;Shin, Hyun-Young;Oqani, Reza;Kim, So-Yeon;Jin, Dong-Il
    • Korean Journal of Agricultural Science
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    • v.43 no.4
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    • pp.575-580
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    • 2016
  • This study was performed to determine whether supplementation of tauroursodeoxycholic acid (TUDCA), an endoplasmic reticulum (ER) stress inhibitor, during vitrified cryopreservation enhances the development of frozen mouse embryos. Mouse 8-cell stage embryos were collected and exposed to a cryoprotectant solution containing TUDCA or TM (tunicamycin, an ER stress inhibitor) at room temperature and stored in liquid nitrogen following vitrification. The final concentration of TUDCA or TM was $50{\mu}M$. The survival and development rates of mouse 8-cell stage embryos exposed to TUDCA- or TM-containing solutions at room temperature or stored in liquid nitrogen following vitrification were measured. There were no significant differences in survival rate and blastocyst formation rate among control, TUDCA, and TM groups after embryos were exposed to vitrification solutions at RT. When mouse 8-cell stage embryos were treated with TUDCA or TM and then stored in liquid nitrogen, the survival rates of control and TUDCA groups were significantly higher than for the TM group. Blastocyst formation rate of the TUDCA group following in vitro culture was significantly higher than that in control or TM groups. The TM group showed a lower (p < 0.05) blastocyst formation rate than the other two groups. Our results indicate that TUDCA supplementation during cryopreservation of mouse embryos could enhance their development capacity.

Essential Oil of Tridax procumbens L Induces Apoptosis and Suppresses Angiogenesis and Lung Metastasis of the B16F-10 Cell Line in C57BL/6 Mice

  • Manjamalai, A.;Kumar, M.J. Mahesh;Grace, V.M. Berlin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5887-5895
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    • 2012
  • Background: To determine the effect of essential oil obtained from a traditionally used medicinal plant Tridax procumbens L, on lung metastasis developed by B16F-10 melanoma cells in C57BL/6 mice. Materials and Methods: Parameters studied were toxicity, lung tumor nodule count, histopathological features, tumor directed capillary vessel formation, apoptosis and expression levels of $P^{53}$ and caspase-3 proteins. Results: In vitro the MTT assay showed cytotoxicity was found to be high as 70.2% of cancer cell death within 24hrs for $50{\mu}g$. In vivo oil treatment significantly inhibited tumor nodule formation by 71.7% when compared with untreated mice. Formation of tumor directed new blood vessels was also found to be inhibited to about 39.5%. TUNEL assays also demonstrated a significant increase in the number of apoptotic positive cells after the treatment. $P^{53}$ and caspase-3 expression was also found to be greater in the essential oil treated group than the normal and cancer group. Conclusions: The present investigation showed significant effects of the essential oil of Tridax procumbens L in preventing lung metastasis by B16F-10 cell line in C57BL/6 mice. Its specific preventive effect on tumor directed angiogenesis and inducing effect on apoptosis warrant further studies at the molecular level to validate the significance of Tridax procumbens L for anticancer therapy.

Study on Chemicals for Post-activation in Porcine Somatic Cell Nuclear Transfer

  • Min, Kyuhong;Na, Seungwon;Lee, Euncheol;Kim, Ghangyong;Yu, Youngkwang;Roy, Pantu Kumar;Fang, Xun;Salih, MB;Cho, Jongki
    • Journal of Embryo Transfer
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    • v.31 no.2
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    • pp.131-136
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    • 2016
  • Since the first success of animal cloning, somatic cell nuclear transfer presented various ideas in many research areas such as regenerative medicine. However, SCNT embryos has poor survival rate. Therefore, numerous researches carried out to enhance the developmental capability of porcine nuclear transfer embryos. Cytochalasin B, demecolcine, latrunculin A, cycloheximide and 6-dimethylaminopurine are efficient chemicals treated in post-activation procedure to increase the efficiency of SCNT. This review study is aim to investigate the effects of these chemicals applied to post-activation in porcine SCNT. Cytochalasin B, demecolcine, latrunculin A are cytoskeletal manuplators inhibit extrusion of pseudo-polar body. Cytochalasin B and demecolcine showed considerably higher blastocyst formation proportion (26-28%) compared to when they are not treated (16%). And when latrunculin A was treated for postactivation, blastocyst formation proportion was increased in SCNT embryos exposed to LA (38%) than those in control (14%). On the other hand, cycloheximide and 6-dimethylaminopurine are protein synthesis and kinase inhibitors. And they help to maintain $Ca^{2+}$ fluctuation in oocytes. Cleavage and blastocyst rates of NT embryos were increased when they were exposed to CHX (16.9% and 5.4% with no CHX).And 6-DMAP also showed higher blastocyst formation (21.5% compared to 15.7%, control). Although all these chemicals have different mechanisms, they showed developmental competence enhancement in NT embryos. However, there are only few studies comparing each chemical's post-activation effect. Therefore, further research and study should be conducted to find optimal chemical for improving the efficiency of SCNT.

Mesodermal Formation and Patterning during Ascidian Embryogenesis (멍게 배발생 과정에서 중배엽 형성과 패턴화)

  • 김길중;니시다히로키
    • Development and Reproduction
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    • v.6 no.2
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    • pp.77-82
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    • 2002
  • In ascidians, a primitive chordate, maternal cytoplasmic factors and inductive interactions are involved in the specification of cell fates in early embryos. The larval structure of ascidians is relatively simple, and the major mesodermal tissues of the tadpole larva are notochord, muscle, and mesenchyme. Formation of muscle cells is a cell-autonomous process, and localized maternal macho-l mRNA specify muscle fate in the posterior marginal zone of the early embryo. In contrast, inductive influence from endoderm precursors plays important roles in the specification of notochord and mesenchyme fates. FGF-Ras-MAPK signaling is involved In the induction of both tissues. The difference in responsiveness of the posterior mesenchyme and anterior notochord precursors to FGF signaling is caused by the presence or absence of intrinsic factors that inherited from the posterior-vegetal egg cytoplasm, respectively. In these inductions, directed signal polarizes the induced cells and promotes asymmetric cell divisions to produce two daughter cells with distinct fates.

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Toxicological Effects of B(a)P on Preimplantation Mouse Embryos in Vitro (in vitro에서 B(a)P이 착상전 마우스 배자에 미치는 독성학적 영향에 관한 연구)

  • 박귀례;이유미;김판기;신재호;강태석;김주일;장성재
    • Journal of Environmental Health Sciences
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    • v.24 no.2
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    • pp.126-133
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    • 1998
  • Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

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Role of OrfQ in Formation of Light-Harvesting Complex of Rhodobacter sphaeroides under Light-Limiting Photoheterotrophic Conditions

  • LIM, SOO-KYONG;IL HAN LEE;KUN-SOO KIM;JEONG KUG LEE
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.604-612
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    • 1999
  • A puc-deleted cell of Rhodobacter sphaeroides grows with a doubling time longer than 160 h under light-limiting photoheterotrophic (3 Watts [W]/㎡) conditions due to an absence of the peripheral light-harvesting B800-850 complex. A spontaneous fast-growing mutant, R. sphaeroides SK101, was isolated from the puc-deleted cells cultured photoheterotrophically at 3 W/㎡. This mutant grew with an approximately 40-h doubling time. The growth of the mutant, however, was indistinguishable from its parental strain during photoheterotrophic growth at 10 W/㎡ as well as during aerobic growth. The membrane of SK101 grown aerobically did not reveal the presence of any spectral complex, while the amounts of the B875 complex and photosynthetic pigments of SK101 grown anaerobiclly in the dark with dimethylsulfoxide (DMSO) were the same as those of the parental cell. These results indicate that the oxygen control of the photosynthetic complex formation remained unaltered in the mutant. The B875 complex of SK101 under light-limiting conditions was elevated by 20% to 30% compared with that of the parental cell, which reflected the parallel increase of the bacteriochlorophyll and carotenoid contents of the mutant. When the puc was restored in SK101, the B875 complex level remained unchanged, but that of the B800-850 complex increased. The mutated phenotype of SK101 was complemented with orfQ encoding a putative bacteriochlorophyll-mobilizing protein. Accordingly, it is proposed that the mutated OrfQ of SK101 should have an altered affinity towards the assembly factor specific to the most peripheral light-harvesting complex, which could be either the B875 or the B800-850 complex.

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Fusion Hybrid and Carotenoid Formation from the Yeast, Phaffia rhodozyma (효모 Phaffia rhodozyma의 융합체와 Carotenoid 생성)

  • Chang, Kee-Myung;Kim, Moon-Hee;Song, Myung-Hee;Kim, Sang-Moon;Chun, Soon-Bai
    • The Korean Journal of Mycology
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    • v.21 no.1
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    • pp.9-15
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    • 1993
  • ABSTRACT: The cell fusants were constructed from complementary auxotrophic mutants of Phffia rhodozyma. The nuclear fusion of the fusants was demonstrated by several techniques including comparision of cell volume, estimation of DNA content and nuclear staining. The cell fusants were very stable for succeeding transfer culture on complex medium for more than one year. Malt extract (1%, w/v) and abscisic acid(1 mM) increased the carotenoid formation whereas gibberellic acid(5mM) and riboflavin(0.1 mM) decreased the corresponding content.

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Study for Possibility of N,N,N-Trimethylphytosphingosine (TMP) for Management of Chronic Skin Diseases (N,N,N-Trimethylphytosphingosine (TMP)의 염증성 피부질환 치료제 가능성에 관한 연구)

  • Seo, Won-Sang;Oh, Han-Na;Park, Woo-Jung;Um, Sang-Young;Kang, Sang-Mo
    • KSBB Journal
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    • v.29 no.1
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    • pp.36-41
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    • 2014
  • Skin disease is one of the most common diseases and its incidence is increasing dramatically in modern society. Specially, many attempts have been made to treat chronic skin inflammation diseases, such as psoriasis and atopic dermatitis, but effective therapies for the immune cell-mediated skin diseases, including psoriasis and atopic dermatitis have not been developed. Until recently, several drug candidates which were claimed to be effective for skin diseases have been reported, but most of them are not used to treat chronic skin disease. Especially, Psoriasis is characterized by excessive growth and aberrant differentiation of keratinocytes, but is fully reversible with appropriate therapy. The trigger of the keratinocyte response is thought to be activation of the cellular immune system, with T cells and various immune-related cytokines. Formation of new blood vessels starts with early psoriatic changes and disappears with disease clearance. Several angiogenic mediators are up-regulated in psoriasis development. Contact- and mediator-dependent factors derived from keratinocytes, mast cells and immune cells may contribute to the strong blood vessel formation of psoriasis. New technologies and experimental models provide new insights into the role of angiogenesis in psoriasis pathogenesis. TMP and its derivatives themselves effectively inhibited in vitro cell migration, tube formation, and the expression of angiogenic factors. However, TMP and its derivatives induced side effects including hemolysis and local side effects. Therefore, in an attempt to reduce the toxicity and the undesirable side effects of TMP and derivatives, a liposomal formulation was prepared and tested for its effectiveness. TMP and derivatives liposomes retained the effectiveness of TMP in vitro while side effects were reduced. These results support the conclusion that TMP effectively inhibits in vitro angiogenesis, with the possibility that use as a psoriasis relief agent.

Characteristics of a Rhodobacter sphaeroides mutant selected by increased growth rate under light-limiting photoheterotrophic conditions

  • Lim, Soo-Kyoung;Lee, Il-Han;Lee, Jeong K.
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.08a
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    • pp.55-67
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    • 1999
  • A puc -deleted cell of Rhodobacter sphaeroides grows with a doubling time longer than 160 h under the light-limiting photoheterotrophic ( 3 Watts [W]/㎡) conditions due to an absence of the peripheral light-harvesting B800-850 complex. A spontaneous fast-growing mutant, R.sphaeroides SK101 was ioslate dto have∼40-h doubling at 3 Watts/㎡, while the growth of the mutant was not distinguished from its parental strain during both aerobic and light-saturating photoheterotrphic (10W/㎡) growth. The B875 complex of SK101 under the light-limiting conditions was elevated by 20 to 30% compared with that of the puc -deleted cell, reflecting parallel increase of bacteriochlorophyll and carotenoid contents of the mutant. The formation of B875 complex of SK101 under the anaerobic dark conditions with dimethylsulfoxide was the same as that of the puc-deleted cell. suggesting that the mutation of SK101 result in the altered control of B875 complex formation by light. When puc is restored in SK101 , it is not B875 complex but B800-850 complex which formation is elevated. The mutation of SK101 affected the bchF transcription most drastically to show two to tenfold increase during both aerobic and photoheterotrophic growth. The mutated phenotype of SK101 was complemented with pW2, which contains approximately 100-kb HNA of the photosynthetic gene clusters. The complementing DNA was narrowed down to a 1.1-kb DNA containing orfQ and pufKBA . The mutation of SK101 appeared to be exerted through the mutation of the orfQ gene encoding a putative bacteriochlorophyll -mobilizing protein.

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