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Oviductal Protein Produces Flurescence Staining of the Perivitelline Space in Mouse Oocytes
KIM, HYUNCHAN,KIM, HAEKWON,KIM, SUNG RYE,KIM, MOON KYOO,SCHUETZ, ALLEN W. 이화여자대학교 생명과학연구소 1996 생명과학연구논문집 Vol.7 No.-
Mouse oocytes were previously observed to undergo structural changes in volving the perivitelline space(PVS)within the oviduct following ovulation, as visualized by staining with fluorochrome-protein conjugates. In the present study, this phenomenon was investigated in detail to determine the role of the oviduct and oocyte. Mouse ovarian oocytes matured in vitro were further incubated in medium or within explanted oviducts in vitro for varying periods of time and then stained with fluorescein isothiocyanate(FITC)-casein. Twenty percent of oocytes incubated within explanted oviducts for 3 hr showed distinct fluorescence staining of PVS, whereas after 20 hr incubation, most(89%)oocytes were similarly stained. In contrast, no ovarian oocytes was stained when incubated in medium alone. Puromycin treatment during incubation of oocytes within explanted oviducts produced a dose-dependent decrease in the percentage of oocytes exhibiting PVS staining after FITC-casein exposure. FITC-casein staining of the PVS also occurred in all oocytes following incubation of in vitromatured oocytes with oviductal tissue extract. In contrast, no oocytes incubated with serum exhibited fluorescence staining. Additionally, the PVS of oocytes failed to stain after incubation with either 0.001% of trypsin- or heat-treated oviductal homogenate. When zona pellucida(ZP)ghosts, devoid of ooplasm, were incubated within explanted oviducts, their PVS was stained brightly following FITC-casein treatment. From these results, it is concluded that proteinaceous material(s)secreted by the mouse oviduct is responsible for the fluorescence staining of the PVS of mouse oocytes and of ghost ZP. The ooplasm does not appear to paly any role inn altering the properties of the PVS staining.
<i>Ex Vivo</i> Characteristics of Human Amniotic Membrane-Derived Stem Cells
Kim, Jiyoung,Kang, Hyun Mi,Kim, Haekwon,Kim, Mee Ran,Kwon, Hyuck Chan,Gye, Myung Chan,Kang, Sung Goo,Yang, H. Seung,You, Juice Mary Ann Liebert 2007 Cloning & stem cells Vol.9 No.4
<P>Cells were isolated from four human amniotic membranes, and their biological characteristics analyzed during ex vivo expansion. Morphologically homogenous populations of fibroblast-like cells were obtained from the second or third passage. Under the appropriate culture conditions, these human amniotic membrane-derived mesenchymal cells (HAM) were shown to differentiate into adipocytes, osteocytes, chondrocytes and neuronal cells, as visualized by Oil Red O, von Kossa, alcian blue, anti-Neu N, and anti-Gal C antibody staining, respectively. Immunophenotype analysis of HAM cells revealed the presence of antigens for SSEA-3, SSEA-4, collagen type-I, -II, -III, -IV, -XII, fibronectin, alpha-SMA, vimentin, desmin, cytokeratin18 (CK18), HCAM-1, fibroblast surface protein, and human leukocyte antigen (HLA) ABC. ICAM-1 protein was weakly detectable, and proteins of TRA-1-60, VCAM-1, von Willebrand factor, PECAM-1, and HLA DR were not detected. HAM cells reached senescence after 14.5+/-0.9 passages, over a period of 146.8+/-8.9 days, and underwent an average of 36.9 4.7 population doublings. RT-PCR analysis showed that all four HAM cell lines consistently expressed genes of Oct-4, Rex-1, SCF, NCAM, nestin, BMP-4, GATA-4, HNF-4alpha, vimentin, and CK18, regardless of the passage number. The genes of Brachyury, FGF-5, Pax-6, and BMP2 were never expressed. Strikingly, alpha-fetoprotein (alphaFP), HLA ABC, and HLA DR genes were expressed in an earlier passage but not expressed in later passages. Telomerase activity of two HAM lines was discernable upon the third passage. These observations strongly suggest that HAM might be immune-privileged and, thus, advantageous as therapeutic cells.</P>
Studies of the Book of Isaiah in the Korean Church since its Beginning
HaeKwon Kim 성공회대학교 신학연구소 2011 Madang: Journal of Contextual Theology Vol.15 No.-
I thank God for letting us KSOTS to celebrate its jubilee anniversary by holding an international conference with the topic “Old Testament Studies in the Era of Globalization,” which deals with two subthemes, “The Old Testament and Its Interpretation in the Cross-cultural World” and “The Use and Interpretation of the Old Testament in Korea and Its Related Society.” I am honored to present a paper at this important conference and want to introduce to participants how the Prophets in general and the Book of Isaiah in particular have been read and studied in the Korean Church and academia.
Kim Minjung,Minjeong Hong,Kim Jisoo,Kim Haekwon,Lee Seung-Jae,Kang Sung-Goo,Cho Dong-Jae 한국발생생물학회 2001 한국발생생물학회 학술발표대회 Vol.2001 No.-
Gelatin zymograms of bFF and bS showed GA110 and 62 kDa gelatinses in adsition to several minor ones. Of these, GA110 gelatinase was abolished by treating bFF or bS with bOF and interestingly, its enzymatic activity was enhanced by adding EDTA to bFF or bS before zymographic analyses. Experiments using specific inhibitors of MMPs indicated that GA110 and 62 kDa proteins were indeed gelatinases. Immunoblotting experiments using an antibody against human MMP-2 showed that both GA110 and 62 kDa were an MMP-2 isoform and active MMP-2, respectively. The results suggest that the interaction between bFF and bOF can occur at the time of fertilization.
Ex Vivo Expansion of Hematopoietic Stem/Progenitor Cells by Coculture using Insert
Kim Kyung-Suk,Kim Haekwon,Do Byung-Rok,Park Seah,Kwon Hyuck-Chan,Kim Hyun-Ok,Im Jung-Ae 한국발생생물학회 2003 한국발생생물학회 학술발표대회 Vol.2003 No.1
Coculture of HSC with bone marrow-derived mesenchymal stem cells (BM-MSCs) is one of used methods to increase cell numbers before transplant to the patients. However, because of difficulties to purify HSCs after coculture with BM-MSCs, it needs to develop a method to overcome the problem. In the present study, we have examined whether a culture insert placed over a feeder layer might support the expansion of HSCs within the insert. cells isolated from the umbilical cord blood by using midiMACS were divided into three groups. A group of 1 cells were grown on a culture insert without feeder layer (Direct). The same number of HSCs was directly cocultured with BM-MSCs (Contact). The third group was placed onto an insert below which BM-MSCs were grown (Insert). To distinguish feeder cells from HSCs, BM-MSCs was pre-labeled fluorescently with PKH26 and 1 cells were seeded in the culture dishes. After culture for 13 days, the expansion factor (x) of HSCs that were grown without feeder layer (Direct) was In contrast, the number of HSCs directly cocultured with feeder layer was 59.6 0.5 and that of HSCs cultured onto an insert was The percentage of BM-MSCs cells remained being fluorescent was after culture. Immune-phenotypically large proportion of cultured cells were founded to be differentiated into myeloid/monocyte progenitor cells. The ability of BM-MSCs, fetal lung, cartilage and brain tissue cells to support ex vivo expansion of HSCs was also examined using the insert. After 11 days of coculture with each of these cells, the expansion factor of HSCs was 15.0, 39.0, 32.0 and 24.0, respectively. Based upon these observations, it is concluded that the coculture method using insert is very effective to support ex vivo expansion of HSCs and to eliminate the contamination of other cells used to coculture wth HSCs.
Kim, Jiyoung,Kim, Haekwon,Lee, Joon Yeong,Choi, Young Min,Lee, Su-Jae,Lee, Seung-Jae CSIRO Publishing 2005 Reproduction, fertility, and development Vol.17 No.5
<P> The aim of the present study was to determine whether a disintegrin and metalloproteinase (ADAM)-8, -9, -10, -12, -15 and -17 and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-1 are involved in the remodelling process of the mouse uterus during the oestrous cycle. The mRNA expression of ADAM was observed in all uterine tissues throughout the entire cycle. The levels of ADAM-8 mRNA were maximal at pro-oestrus, whereas the expression of ADAM-9 and ADAMTS-1 mRNA was maximal at oestrus. The minimum mRNA level of all ADAM genes always occurred at dioestrus. The mRNA levels of ADAM-10, -12, -15 and -17 did not vary significantly, regardless of the stage of the oestrous cycle. Immunoblot analyses demonstrated the presence of all ADAM proteins throughout the cycle. In terms of protein intensities, ADAM-8, -12 and -17 were maximal at pro-oestrus, whereas ADAM-10 and ADAMTS-1 were maximal at metoestrus and ADAM-9 was maximal at oestrus. Regardless of the ADAM species, minimal protein expression always occurred at dioestrus. Immunohistochemical studies showed ADAM protein expression in luminal and glandular epithelial layers, but not in the stromal layer. Moreover, ADAM proteins were found to be heterogeneously localised and their individual localisations depended on the stage of the oestrous cycle. From these observations, we suggest that the ADAM genes play an important role in mouse uterine tissue remodelling during the oestrous cycle. </P>
Establishment of Stem-like Cells from Human Umbilical Cord Vein
Park Seah,Kim Kyung-Suk,Kim Haekwon,Do Byung-Rok,Kwon Hyuck-Chan,Kim Hyun-Ok,Im Jung-Ae 한국발생생물학회 2003 한국발생생물학회 학술발표대회 Vol.2003 No.1
Adult stem cells can make identical copies of themselves for long periods of time. They also give rise to many differentiated mature cell types that have characteristic morphology and specialized function. Human adult stem cells are the attractive raw materials for the cell/tissue therapy, however, it is not easy to get from the adult tissues. In the present study, we tried to isolate a cell population derived from human umbilical cord vein which has been discarded after birth. The cells were isolated after treatment of the umbilical vein with collagenase or trypsin. After 3 days of culture, two kinds of cell populations were found consisting of adherent cells with endothelial cell-like and fibroblast-like morphology, respectively. When these cells were subcultured 12 times over a period of 3 months, almost cells appeared uniformly to exhibit fibroblastoid morphology which was different from that of mesenchymal stem cells obtained from human bone marrow The results of RT-PCR analyses showed distinct expression of BMP-4, oct-4, and SCF genes but not of GATA, PAX-6 and Brachyury genes. On immunohistochemical staining, the cells were negative for the von Willebrand factor(vWF), alpha-smooth muscle actin and placental alkaline phosphatase. From these observations, it is suggested that stem-like cells might be present in human umbilical cord vein.