• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.10a
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• pp.49-49
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• 2005

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2007.07a
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• pp.6-7
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• 2007

• Development and Reproduction
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• v.11 no.2
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• pp.59-66
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• 2007

Understanding molecular mechanisms that control embryonic stem cell (ESC) self-renewal and differentiation is important for the development of ESC-based therapies. Statins, inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), potently reduce cholesterol level. As well as inhibiting cholesterol synthesis, statins inhibit other intermediates in the mevalonate pathway such as farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP), major substrates for protein isoprenylation. Studies showed that pleiotropic effects of statins beyond cholesterol lowering property arise from inhibition of protein isoprenylation that is involved in various cellular functions including proliferation and differentiation. It has been determined that statins have inhibitory effect on ESC self-renewal and stimulatory effect on ESC differentiation into adipogenic/osteogenic lineages. Importantly, statins mediate downregulation of ESC self-renewal by inhibiting RhoA-dependent signaling, independently of their choresterol-lowering properties. Understanding statin's actions on ESCs may provide important insights into the molecular mechanisms that regulate self-renewal or differentiation of ESCs.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.550-556
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• 1995

The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.

• Development and Reproduction
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• v.12 no.1
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• pp.87-95
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• 2008

Neural stem/precursor derived from pluripotent human embryonic stem cells (hESCs) has considerable therapeutic potential due to their ability to generate various neural cells which can be used in cell-replacement therapies for neurodegenerative diseases. However, production of neural cells from hESCs remains technically very difficult. Understanding neural-tube like rosette characteristic neural precursor cells from hESCs may provide useful information to increase the efficiency of hESC neural differentiation. Generally, neural rosettes were derived from differentiating hEBs in attached culture system, however this is time-consuming and complicated. Here, we examined if neural rosettes could be formed in suspension culture system by bypassing attachment requirement. First, we tested whether the size of hESC clumps affected the formation of human embryonic bodies (hEBs) and neural differentiation. We confirmed that hEBs derived from $500{\times}500\;{\mu}m$ square sized hESC clumps were effectively differentiated into neural lineage than those of the other sizes. To induce the rosette formation, regular size hEBs were derived by incubation of hESC clumps($500{\times}500\;{\mu}m$) in EB medium for 1 wk in a suspended condition on low attachment culture dish and further incubated for additional $1{\sim}2$ wks in neuroectodermal sphere(NES)-culture medium. We observed the neural tube-like rosette structure from hEBs after $7{\sim}10$ days of differentiation. Their identity as a neural precursor cells was assessed by measuring their expressions of neural precursor markers(Vimentin, Nestin, MSI1, MSI2, Prominin-1, Pax6, Sox1, N-cadherin, Otx2, and Tuj1) by RT-PCR and immunofluorescence staining. We also confirmed that neural rosettes could be terminally differentiated into mature neural cell types by additional incubation for $2{\sim}6$ wks with NES medium without growth factors. Neuronal(Tuj1, MAP2, GABA) and glial($S100{\beta}$ and GFAP) markers were highly expressed after $2{\sim}3$ and 4 wks of incubation, respectively. Expression of oligodendrocyte markers O1 and CNPase was significantly increased after $5{\sim}6$ wks of incubation. Our results demonstrate that rosette forming neural precursor cells could be successfully derived from suspension culture system and that will not only help us understand the neural differentiation process of hESCs but also simplify the derivation process of neural precursors from hESCs.

• Journal of the Korean Society of Radiology
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• v.85 no.4
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• pp.813-819
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• 2024

Ectopic breast tissue, which results from incomplete regression of the mammary line during embryogenesis, is observed in 0.2%-6% of the population. Carcinoma development in ectopic breast tissue, especially in the abdominal or chest wall, is rare. Here we present the unusual case of a 38-year-old woman with invasive ductal carcinoma in the ectopic breast tissue on the left side of the chest wall and concurrent fibroadenoma in the ectopic breast tissue on the right side. We also describe the US and MR findings of these masses.

• Development and Reproduction
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• v.12 no.1
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• pp.1-8
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• 2008

Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

• The Korean Journal of Zoology
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• v.15 no.1
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• pp.25-33
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• 1972

Two-Cell mouse embryos were incubated in the anterior chamber of the rat eye, which has been known as the best place among other animals' for the mouse ovum maturation, in order to observe the capability of their early development. Within 120 hours after incubation, 71.0% of two-cell embryos have developed to the blastocysts in the male rat eye, while only 38.5% in the eye of the same mouse as donated two-cell embryos. Thus, the rat eye chamber provides more favourable environment to the embryos than the mouse itself. The results are consistent with those of the previous studies comparing the maturation of the mouse follicular oocytes in the mouse and the rat eye chamber. Although the aqueous humor which is filled in the anterior chamber of the eye is characterized by its specific properties, being of higher osmolarity, higher concentrations of ascorbic acid, pyruvate and lactate, but lower of proteins and lower temperature than those in blood or lymph serum, The embryos are able to under-take their cleavage as normal as in vivo or in vitro. Concerning with a number of studies in vitro on the development of the mouse embryos which are requiring a very limited condition, the fact that they are able to manage their further development under very different enviroment from our knowledges would provide us a moment to understand their behavior during the early development. The difference of the proportion of the developed blastocysts between in the mouse eye chamber and in the rat can possibly be resulted from the species specific difference in the physicochemical properties between their eye chambers. This assumption is based upon the findings by many investigators who chmpared the nature of the eye chamber of various animals. As a consequence, the rat eye chamber might consist of better properties for the embryonal growth than the mouse eye chamber. The mouse embryos cleaved with a delayed period. In normal development they complete almost the cleavage within 94 hours after fertilization. However, in the present studies, 81.1% of two-cell embryos developed to the blastocysts and the morula in 120 hours in the eye chamber, assumed to be about 154 hours after fertilization. Such delay in development would be caused mainly by the low temperature of the eye chamber. At present we can make two assumptions to explain the capability of the emtryonal development in the eye chambers. One is that the embryos would possess an ability to adapt themselves to the environment which provides unfavourable conditions. The other is that the embryos might remain for a certain duration in the eye chamber, which is filled with a new body fluid produced immediately after the loss of the aqueous humor and the fluid of which becomes similar to blood serum in component. The first assumption is highly reliable since the embryonal cells are mostly at the undifferentiated state and so they probably engage a simple metabolism during their early period. The second assumption is induced by the fact that the rabbit eye chamber produces a plasmoid humor which has mostly similar components to blood serum after loss of aqueous humor through cornea by puncturing. However, the plasmoid humor is substituted by the initial aqueous humor in eight hours. Even though this finding, production of the new fluid, could be applied to the rat eye, it is hardly reliabel that the plasmoid humor remains for such a long period as 120 hours. Consequently, the development of the embryos is more likely due to their adaptability to the new environment during their early developmental stages.

• Development and Reproduction
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• v.11 no.1
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• pp.21-29
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• 2007

Previously, we found progesterone receptor membrane component 2 (pgrmc2) was highly expressed in germinal vesicle (GV) stage oocytes. The present study was conducted to characterize the expression of pgrmc2, as well as pgrmc1, in the mouse gonads and embryos according to their developmental stages. We found that these membrane components were expressed in ovaries, testes, and embryos at various developmental stages in addition to oocytes. Progesterone-3-O-carboxymethyl oxime-BSA-fluorescein isothiocyanate (P4-BSA-FITC) was applied to visualize the presence of the progesterone receptor on mouse oocyte membrane, and we confirmed that immobilized progesterone is localized at surface of the oocyte. This is, at our knowledge, the first report regarding the expression of membrane component of progesterone receptor in the mouse oocytes, embryos, and gonads. The function and signal transduction pathway of progesterone receptor membrane components in oocytes requires further studies.

• Development and Reproduction
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• v.14 no.2
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• pp.43-50
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• 2010

Recently induced pluripotent stem (iPS) cells are derived from somatic cells by ectopic expression of several transcription factors (reprogramming factors) using technology of somatic cell reprogramming. iPS cells are able to selfrenew and differentiate into all type of cells in the body similarly to embryonic stem cells. Because iPS cells have advantages that can avoid immune rejection after transplantation and ethical issues unlike embryonic stem cells, research on iPS has made significant progress since the first report by Yamanaka in 2006. Nevertheless of many advantages of iPS, safer methods to introduce reprogramming factors into somatic cells must be developed due to safety concerns regarding viral vectors, and safer reprogramming factors to substitute the oncogenes should be evaluated for clinical application of iPS. Here we discuss the recent progress in reprogramming factors in embryonic stem cells, oocytes, and embryos, and discuss further research for finding new, more reliable and safer reprogramming factors.