• Title/Summary/Keyword: Embryonic induction

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Ovulation Induction Effect of Sevenband Grouper, Epinephelus septemfasciatus by Treating Hormones (호르몬 처리에 의한 능성어 Epinephelus septemfasciatus의 배란유도 효과)

  • HONG, Chang-Gi;CHO, Jae-Kwon;PARK, Jong-Youn;SON, Maeng-Hyun;PARK, Jae-Min;HAN, Kyeong-Ho;KANG, Hee-Woong
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.4
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    • pp.981-989
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    • 2015
  • This study aims to investigate how ovulation induction of sevenband grouper, Epinephelus septemfasciatus varies when treated with different hormonal substances such as ovaprim, pimozide, LHRHa, and HCG. As for LHRHa, we injected it in different concentrations: $50{\mu}g/kg$, $100{\mu}g/kg$, $150{\mu}g/kg$, and $200{\mu}g/kg$. All hormonal substances including LHRHa were injected into back muscles. As a result, ovulation occurred in all sample groups. The sample group injected with HCG released the largest amount of eggs, and the dimeter of eggs and oil globules were also significantly bigger than those of others. Fertilization rate, embryonic survival rate, and hatching rate were the highest in a LHRHa ($100{\mu}g/kg$) +pimozide ($1,000{\mu}g/kg$) group and buoyant rate in a LHRHa group. Ovulation rate was 100% when LHRHa was injected in different concentrations. The largest amount of eggs was released in $200{\mu}g/kg$. The diameter of eggs and oil globules were the biggest in $200{\mu}g/kg$ and the smallest in $50{\mu}g/kg$. Fertilization rate showed no significant difference through all different levels of concentration: 94.6-95.3%. Buoyant rate was the highest in $100{\mu}g/kg$ and embryonic survival rate and hatching rate in $150{\mu}g/kg$. Based on these experimental results, we concluded that LHRHa in $100{\mu}g/kg$ is the most efficient for the ovulation induction of sevenband grouper.

Establishment of Mouse Embryonic Stem Cell and Effects of Herbal Medicine on Induction of Cardiomyocyte Differentiation

  • Lee, Ji Hyang;Lee, Eun
    • Korean Journal of Plant Resources
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    • v.25 no.6
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    • pp.693-699
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    • 2012
  • This study was conducted to investigate the effects of Woohwangcheungsimweun (ox bezoar), deer antlers, and wild ginseng on induction of cardiomyocyte differentiation using the established mouse embryonic stem (ES) cells. The expression of atrial natriuretic peptide (ANP) was highest in Woohwangcheungsimweun treatment group. The expression of rabbit anti-GATA-4(GATA-4) and troponin (TnI) were highest in wild ginseng and Woohwangcheungsimweun treatment groups, respectively. Fluorescence activated cell sorting (FACS) analysis showed that the expression of ANP was highest in Dimethyl sulfoxide(DMSO) and Woohwangcheungsimweun treatment groups. The expression of GATA-4 was relatively high in wild ginseng treatment group. The expression of TnI was highest in Woohwangcheungsimweun treatment group. In the gene expression analysis, DMSO greatly inhibited GATA-4 expression to 25% of control. Woohwangcheungsimweun treatment caused to increase cTnI and cardiac ANP expression significantly. Wild ginseng extract upregulated GATA-4 gene expression. In conclusion, DMSO widely used as cardiomyocyte differentiation inducer did not show significant effects on the expression of ANP, GATA-4 and TnI in this study. Woohwangcheungsimweun showed upregulation of ANP and TnI expression. Wild ginseng extract showed greater effects than DMSO on GATA-4 expression. These results might suggest that the combination of Woohwangcheungsimweun and wild ginseng extract treatment can be expected to increase expressions of all three genes.

Deficiency of Bloom's Syndrome Protein Causes Hypersensitivity of C. elegans to Ionizing Radiation but Not to UV Radiation, and Induces p53-dependent Physiological Apoptosis

  • Kim, Yun Mi;Yang, Insil;Lee, Jiyeung;Koo, Hyeon-Sook
    • Molecules and Cells
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    • v.20 no.2
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    • pp.228-234
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    • 2005
  • Caenorhabditis elegans him-6 mutants, which show a high incidence of males and partial embryonic lethality, are defective in the orthologue of human Bloom's syndrome protein (BLM). When strain him-6(e1104) containing a missense him-6 mutation was irradiated with ${\gamma}$-rays during germ cell development or embryogenesis, embryonic lethality was higher than in the wild type, suggesting a critical function of the wild type gene in mitotic and pachytene stage germ cells as well as in early embryos. Even in the absence of ${\gamma}$-irradiation, apoptosis was elevated in the germ cells of the him-6 strain and this increase was dependent on a functional p53 homologue (CEP-1), suggesting that spontaneous DNA damage accumulates due to him-6 deficiency. However, induction of germline apoptosis by ionizing radiation was not significantly affected by the deficiency, indicating that HIM-6 has no role in the induction of apoptosis by exogenous DNA damage. We conclude that the C. elegans BLM orthologue is involved in DNA repair in promeiotic cells undergoing homologous recombination, as well as in actively dividing germline and somatic cells.

Differentiation of Mesenchymal Stem Cell-like Cell from Feeder Free Cultured Human Embryonic Stem Cells using Direct Induction System (Feeder-free에서 배양된 인간배아줄기세포의 직접분화유도 방법을 이용한 간엽줄기세포로의 분화)

  • Lee, Min-Ji;Lee, Jae-Ho;Kim, Ju-Mi;Shin, Jeong-Min;Park, Soon-Jung;Chung, Sun-Hwa;Lee, Kyung-Il;Chae, Jung-Il;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.1-6
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    • 2010
  • Mesenchymal stem cells (MSCs) have the multipotent capacity and this potential can be applied for obtaining valuable cell types which can use for cell therapy on various regenerative diseases. However, insufficient availability of cellular source is the major problem in cell therapy field using adult stem cell sources. Recently, human embryonic stem cells (hESCs) have been highlighted to overcome a limitation of adult cellular sources because they retain unlimited proliferation capacity and pluripotency. To use of hESCs in cell therapy, above all, animal pathogen free culture system and purification of a specific target cell population to avoid teratoma formation are required. In this study, we describe the differentiation of a mesenchymal stem cell-like cells population from feeder-free cultured hESCs(hESC-MSCs) using direct induction system. hESC-MSCs revealed characteristics similar to MSCs derived from bone marrow, and undifferentiated cell markers were extremely low in hESC-MSCs in RT-PCR, immunostaining and FACS analyses. Thus, this study proffer a basis of effective generation of specialized human mesenchymal stem cell types which can use for further clinical applications, from xenofree cultured hESCs using direct induction system.

Optimization of Human Embryonic Stem Cells into Differentiation of Dopaminergic Neurons in Vitro: I. Additive Effect of Neurotrophic Factor on Human Embryonic Stem Cells

  • 이금실;김은영;이영재;신현아;조황윤;이훈택;정길생;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.79-79
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    • 2003
  • Embryonic stem cells are capable of differentiating into a variety of cell lineages. However, the ultimate results of differentiation in vitro greatly depend on the duration of treatment and kinds of differentiating inducers added. In order to investigate the efficiencies of various differentiation inducers and the methods of treatment, we examined differentiation patterns of human embryonic stem cell (hESC, MB03) according to several different protocols. Exp. I) Upon differentiation using retinoic acid and ascorbic acid (RA/AA), embryoid bodies (EB, for 4days) derived from hESC was exposed to Rh (10$^{-6}$ M) and AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. Exp. III) In addition, to examine the effects of neurotrophic factors in the production of mature neurons, groups of cells were exposed to either BDNF (5 ng/ml) or TGF-$\alpha$(10 ng/ml) during the 28 days of final differentiation. Differentiation patterns of RA/AA or bFGF treated groups were very similar; approximately 82% and 83% of the cells, respectively, were positive for anti-NF200 antibody, while it was about 10% and 11%, respectively, for anti-NF160 antibody in 28 days in N2 medium. Alsor, cells expressing TH were as low as 5%, while the cells doubled when matured at the presence of either BDNF or TGF-$\alpha$. Cells immunoreactive to anti-GAD antibody were approximately 20%. These results suggest that a maturation step rather than differentiation induction step, which is formation of EB, effects more decisively to the ultimate differentiation pattern.

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Differentiation of mouse embryonic stem cell into smooth muscle cells by DBcAMP and retinoic acid (DBcAMP와 retinoic acid를 이용한 마우스 배아줄기의 평활근세포 분화)

  • Park, Sung-Soo;Kang, Ju-Won
    • Korean Journal of Veterinary Service
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    • v.31 no.4
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    • pp.449-456
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    • 2008
  • The differentiation of mouse embryonic stem(ES) cell into smooth muscle cells(SMC) may play a major role in cardiovascular development and under pathophysiological conditions. Therefore, in the present study, we have examined the differentiation of ES cells and its related gene expression. SMC differentiation was indicated by cellular morphology and time-dependent induction of dibutyryl adenosine 3,5-cyclic monophosphate(DBcAMP)and retinoic acid(RA) on smooth muscle ${\alpha}$-actin($SM{\alpha}A$), smooth muscle myosin heavy chain(SMMHC) gene expression. The control was undifferentiated ES cells(protein expressions represent 50-60kDaOct-4). The results of this study show that morphology of embryoid body and confirmation of $SM{\alpha}A$ expression by immunocytochemistry. Moreover, SMMHC and desmin expression was significantly increased by time dependent manner(5, 7, 15 days), in contrast to $SM{\alpha}A$ expression was slightly decreased on 15days. In conclusion, DBcAMP and RA stimulate mouse ES cells differentiation into SMC and enhanced $SM{\alpha}A$, SMMHC and desmin expression.

PRODUCTION, EVOLUTION AND REPRODUCTIVE ENDOCRINOLOGY OF DUCKS

  • Tanabe, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.5 no.1
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    • pp.173-181
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    • 1992
  • Duck is an important domestic animal, especially in Asia. Eighty five percent of ducks in the world are kept in Asia, especially in the East and South Asia regions. The ancestor of domesticated ducks was mallard (Anas platylhynchos), which are still migrating between north and southern parts in Eurasia. Ducks have been domesticated in China for at least 3000 years ago. Phylogenetic studies on ducks, employing electrophoresis of blood proteins, indicate a marked difference of genetic constitution between duck breeds in southeast Asia and those in northeast Asia. Duck embryonic ovary is much more active in secretion of sex steroid hormones especially estradiol than the embryonic testes. Estradiol secreted by the embryonic left ovary has an important role in female sexual differentiation in ducks. In the female ducks, plasma LH, estradiol and testosterone levels increase and reach peaks shortly before the first egg, while progesterone level reach a peak shortly after the first egg. In laying ducks oviposition mostly occurs in the last 3 hr of darkness and first hr of light ranging 02:00-06:00 under 14 hr light (05:00-19:00) and 10 hr darkness photoperiodic condition. Measurements of plasma hormone levels reveal that onset of darkness is a major signal for LH release from the pituitary and the subsequent release of progesterone from ovary, and for induction of ovulation in the female duck.

Effect of Retinoic Acid and dibutyryl cyclic AMP on G1 Phase Associated Molecules during F9 Embryonic Carcinoma Cell Differentiation (Retinoic acid와 dibutyryl cyclic AMP가 F9 embryonic carcinoma cell 분화 중 G1 Phase 관련 분자에 미치는 영향)

  • 박귀례;김건홍;한순영;이유미;장성재
    • YAKHAK HOEJI
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    • v.43 no.3
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    • pp.378-384
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    • 1999
  • Retinoic acid (RA) and dibutyryl cyclic AMP (dbcAMP) induce the differentiation of the multipotent embryonic carcinoma cell line, F9 cells, into parietal endoderm like cell. The F9 cells are highly proliferative doubling approximately 12 hourse. S Phase is predominant, lasting 10 hours and G2/M phase occupies most of the remaining cycle (2 hours) and G1 phase is nearly non-existent. In this study, we showed the effect of RA and dbcAMPon the cell cycle associated molecules (especially around G1 phase) during F9 cell differentiation. Differentiation of F9 cells was induced by the combined addition of RA ($10^{-7}M$) and dbcAMP (0.5mM), and cells were harvested daily up to 4 days. Flow cytometric analysis showed the prolongation of G1 phase around 30 hours after induction. Western blot analysis revealed that the amount of cyclin D1 and cdk2 were increased at day 4. However, histone H1 kinase activity of cdk2 was decreased. These data strongly suggest that RA and dbcAMP induce the growth arrest of F9 cells at G1 phase by decreasing the activity of cdk2, although they have increased the protein contents of cyclin D1 and cdk2. The reason for the discrepancy between the H1 kinase activity and protein contents are not clear yet.

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Studies on the Induction of Transformation in Cereal Plants. III. Cultures and Regeneration of Rice Protoplasts Transferred Foreign Genes. (곡물류의 형질전환 유도에 관한 연구 III. 외래 유전자가 도입된 벼 원형질체의 배양 및 재분화)

  • Hwang, Baik;Hwang, Sung-Jin;Im, Hyong-Tak;Kang, Young-Hee
    • KSBB Journal
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    • v.8 no.1
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    • pp.62-68
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    • 1993
  • Transformed rice plantlet were recovered from protoplasts by electroporation with the plasmld pB 1121, which contain the plant expressible NPT-II and GUS genes. Embryonic cell suspension culture was established with embryonic callus induced from mature seeds of rice (Oryza sativa L. cv. Dong-jin) on the MS medium supplemented with 2.0 mg/l 2,4-D, 0.5 mg/l kinetin, 3% sucrose. Protoplasts isolated from embryonic cell suspensions were electroplated and then poterltialty-transformed tissues were selected by growth on the medium containing 200 mg/l kanamycin sulfate. When subjected to GUS assay, they stained blue, indicating the expression of the inserted GUS genes. Plantlets were regenerated from electroplated protoplasts on the hormone free MS medium. Transferred foreign genes in the plants were confirmed by southern hybridization. These results support use of electroporation for transformation of these important cereal plants.

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