분획용 혈장 마스터 파일 작성지침

강혜나,오영훈,오호정,김판순,김순남,이내리,김정근,박희영,정재호,김주일,홍성화 한국에프디시규제과학회(구 한국에프디시법제학회) 2009 FDC법제연구 Vol.4 No.2

Vero Cell-Conditioned Medium에서의 배양이 생쥐 후기 2-세포기 배아 발생에 미치는 영향

강혜나,하종식 梨花女子大學校 醫科大學 醫科學硏究所 1996 EMJ (Ewha medical journal) Vol.19 No.4

본 실험결과 Vero cell과의 공동배양과 Vero cell-conditioned medium에서의 배양은 생쥐 후기 2-세포기 배아의 부화율을 놓이고 세편이 적은 양질의 배아획득을 돕고 세포 증식을 빠르게 해 팽윤 배반포시 할구수를 증가시키는 것으로 나타났다. 그러나 앞으로 공동배양 기작에 작용할 것으로 보이는 embryotrophic factor나 배양액에서 공동배양 세포주에의해 제거되는 발생저해 물질 등의 분자생물학적 연구가 진행되어야하고, 수정시 정자등이 공동배양 세포주의 monolayer에 결합하여 수정 기회를 감소시켜 여러 장점에도 불구하고 오히려 수정률을 감소시킬 수 있는 적접적인 공동배양보다 conditioned medium에서의 배양에 관심을 가져야할 것으로 본다. This study was conducted to evaluate the ability of Vero cell-conditioned medium for supporting mouse embryo development in vitro. The mouse late 2-cell embryos were cultured in control media(Ham's F-10 +10%FBS), media with Vero cell monolayer and Vero cell-conditioned media for 4 days and measured the hatching rate and cell number in the blastocyst stage. The hatching rate in experimental groups was increased significantly compared with embryos in control group(p<0.01). On the other hand, the degree of blastomere fragmentaion exhibited a opposite trend to that of the developmental capacity(p<0.05). And also the cell numbers of expanded blastocysts in experimental groups were increased significantly compared with the control group(p<0.001). There was, however, no difference between experimental groups. These results indicate that Vero cell-conditioned medium supported the mouse embryo development as a Vero cell monolayer. And the mechanism for enhancement of the development potential of embryos may be releasing the embryotrophic factor during the medium-conditioning period.

생쥐 난자 배양시 외분비 관련 요소들이 자발적 투명대 경화 현상에 미치는 영향

강혜나,배인하,김해권,Kang, Hye-Na,Bae, In-Ha,Kim, Hae-Kwon 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.1

"Spontaneous" hardening of the zona pellucida of mouse oocytes during in vitro culture is most likely due to cortical granules exocytosis. Thus the purpose of the present study was to determine whether the exocytosis factor is involved in spontaneous zona pellucida hardening during in vitro culture of the mouse. The results obtained form these experiments were summarized as follows; 1. When a protein synthesis inhibitor(100${\mu}g$/ml puromycin) was added to the culture medium, it did not prevent spontaneous ZPH of mouse oocyte during in vitro culture. 2. Calmodulin antagonists (trifluoperazine and chlorpromazine) and calcium channel blocker (verapamil) had no inhibitory effect in spontaneous ZPH. 3. A microtubule assembly inhibitor, colcemid had some inhibitory effect on spontaneous ZPH. 4. Treatment with a microfillament formation blocker(cytochalasin-B) at 1${\mu}g$/ml concentration, resulted in the excellent inhibitory effect on spontaneous ZPH. However cytochalasin-B did not inhibit ethanol-induced ZPH.

생쥐 난자 배양시의 자발적 투명대 경화 현상과 PMA에 의한 투명대 경화 현상 비교

강혜나,배인하,김해권,Kang, Hye-Na,Bae, In-Ha,Kim, Hae-Kwon 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.1

One consequence of fertilization in mammals is an increased resistance of the zona pellucida (ZP) to proteases and various chemical reagents. This phenomenon has been called 'zona pellucida hardening' (ZPH), and it is generally accepted that it is caused by the secretory products of cortical granules released by the egg at fertilization. ZP of mouse oocytes maturing in vitro in a chemically defined medium becomes progressively more resistant to solubilization by chymotrypsin ("Spontaneous" ZP hardening). In the present study, it was aimed to find the specificity of spontaneous ZPH in relation to its possible relevance to the cortical reaction and the physiological block to polyspermy. When a maturation inhibitors, cAMP analog(dbcAMP) and phosphodiesterase inhibitor (IBMX) was added to culture medium, it prevent spontaneous ZPH of mouse oocyte during in vitro culture. Thus spontaneous ZPH requires GVBD, since it is prevented by those agents, which inhibit GVBD in vitro. However, culture for 3 hours in the presence of PMA(lOng/ml), a protein kinase C activator, resulted in ZPH without GVBD, thus suggesting that ZPH may be regulated independently apart from the event of GVBD. Pretreatment of mouse oocyte with FBS result in partially inhibitory effect on subsequent spontaneous ZPH. Induction of GVBD in vivo had a inhibitory effect on the spontaneous ZPH, but subsequent spontaneous ZPH. Induction of GVBD in vivo had a inhinbitory effect on the spontaneous ZPII, but had no inhibitory effect on PMA-induced ZPH. Treatment with a microfilament formation blocker(cytochalasin-B) at 1${\mu}g$/ml concentration, resulted in the excellent inhibitory effect on spontaneous ZPH. However cytochalasin-B did not inhibit PMA-induced ZPH. Thus this suggesting that spontaneuse ZPH had a different mechanism from PMA-induced ZPH.

Vero Cell과의 공동배양이 체외에서 생쥐 난자성숙과 배아발생에 미치는 영향

강혜나,하종식 梨花女子大學校 醫科大學 醫科學硏究所 1996 EMJ (Ewha medical journal) Vol.19 No.3

Assisted reproductive technology(ART) have contributed significantly to alleviating subfer-tility in the childless couple. In spite of the many advances in the field of ART, the pregnancy and take-home baby rates for in vitro fertilization(IVF) have been very poor. In order to overcome these problems, a variety of coculture systems has been devised. Vero cells, derived from African green monkey kidney, were selected because kidney and genital tract have a common embryonic origin. In addition, these cells are safe for coculture with embryos : they are highly controlled for viruses and other contaminants because they are used for vaccine production. Several investigators showed that cocultureing human embryos with Vero cells in vitro resulted in an improvement of embryo development. However, they did not observe the same results using mouse oocytes and embryos. We thus designed a series of experiments to demonstrate whether or not Vero cells do indeed enhance mouse oocyte maturation and embryo development. In this experiment, Vero cell does not allow the mouse immature oocytes to be enhanced maturation rate in vitro. To study the 'In-Vitro 2-cell Block' in mouse embryo, we have cocultured ICR one-cell mouse embryos with Vero cell in different medium. In Ham's F-10 the mouse embryos arrested their development prior to 4-cell stage(control 76.7%;coculture 75.0%). In contrast, the coculturing mouse embryos revealed enhanced development(control 0%;coculture 22.8%) in human tubal fluid(HTF) only in late embryonic stages(hatching). On the other hand, the degree of blastomere fragmentation exhibited a reverse trend to that of the developmental capacity. Embryos from coculture groups(Ham's F-10 & HTF) showed some fragmentation(0% & 4.2%) while 13.3% and 14.3% of the embryos in control groups(Ham's F-10 & HTF) were severely fragmented(P<0.05). Thus the use of coculture systems appears to be dependent on the type of medium used as a support. The development rate of late 2-cell mouse embryos in Vero cell coculture was no significant.

Vero Cell과의 공동배양이 체외에서 생쥐 난자성숙과 배아발생에 미치는 영향

강혜나,하종식 이화여자대학교 생명과학연구소 1996 생명과학연구논문집 Vol.7 No.-

Assisted reproductive technology(ART)have contributed significantly to alleviating subfertility in the childless couple. In spite of the many advances in the field of ART, the pregnacy and take-home baby rates for in virto fertilization(IVF)have been very poor. In order to overcome these problems, a variety of coculture systems has been devised. Vero cells, derived from African green monkey kidney, were selected because kidney and genital tract have a common embryonic origin. In addition, these cells are safe for coculture with embryos:they are highly controlled for viruses and other contaminants because they are used for vaccine production. Several investigators showed that coculturing human embryos with Vero cells in vitro resulted in an improvement of embryo development. However, they did not observe the same results using mouse oocytes and embryos. We thus designed a series of experiments to demonstrate whether or not Vero cells do indeed enhance mouse oocyte maturation and embryo development. In this experiment, Vero cell does not allow the mouse immature oocytes to be enhanced maturation rate in vitro. To study the 'In-Vitro 2-Cell Block' in mouse embryo, we have cocultured ICR one-cell mouse embryos with Vero cell in different medium. In Ham's F-10 the mouse embryos arrested their development prior to 4-cell stage(control 76.7%:coculture 75.0%). In contrast, the coculturing mouse embryos revealed enhanced development(control 0%;coculture 22.8%) in human tubal fluid(HTF)only in late embryonic stages(hatching). On the other hand, the degree of blastomere fragmentation exhibited a reverse trend to that of the developmental capacity. Embryos from coculture groups(Ham's F-10 & HTF)showed some fragmentation(0% & 4.2%)while 13.3% and 14.3% of the embryos in control groups(Ham's F-10 & HTF)were severely fragmented (P<0.05). Thus the use of coculture systems appears to be dependent on the type of medium used as a support. The development rate of late 2-cell mouse embryos in Vero cell coculture was no significant differences until blastocyst stage but improved at late develpmental stage(control 42.1% : coculture 70.7%). Thus the Vero cell coculture system was shown to increase the hatching rate of mouse embryos.

Vero Cell-Conditioned Medium에서의 배양이 생쥐 후기 2-세포기 배아 발생에 미치는 영향

강혜나,하종식 이화여자대학교 생명과학연구소 1996 생명과학연구논문집 Vol.7 No.-

본 실험결과 Vero cell과의 공동배양과 Vero cell-conditioned medium에서의 배양은 생쥐 후기 2-세포기 배아의 부화율을 높이고 세편이 적은 양질의 배아획득을 돕고 세포 증식을 빠르게 해 팽윤 배반포시 할구수를 증가시키는 것으로 나타났다. 그러나 앞으로 공동배양 기작에 작용할 것으로 보이는 embryotrophic factor나 배양액에서 공동배양 세포주에 의해 제거되는 발생저해 물질 등의 분자생물학적 연구가 진행되어야 하고, 수정시 정자등이 공동배양 세포주의 monolayer에 결합하여 수정 기회를 감소시켜 여러 장점에도 불구하고 오히려 수정률을 감소시킬 수 있는 직접적인 공동배양보다 conditioned medium에서의 배양에 관심을 가져야할 것으로 본다. This study was conducted to evaluate the ability of Vero cell-conditioned medium for supporting mouse embryo development in vitro. The mouse late 2-cell embryos were cultured in control media(Ham's F-10+10%FBS), media with Vero cell monolayer and Vero cell-conditioned media for 4 days and measured the hatching rate and cell number in the blastocyst stage. The hatching rate in experimental groups was increased significantly compared with embryos in control group(p<0.01). On the other hand, the degree of blastomere fragmentation exhibited a opposite trend to that of the developmental capacity(P<0.05). And also the cell numbers of expanded blastocysts in experimental groups were increased significantly compared with the control group(p<0.001). There was, however, no difference between experimental groups. These results indicate that Vero cell-conditioned medium supported the mouse embryo development as a Vero cell monolayer. And the mechanism for enhancement of the development potential of embryos may be releasing the embryotrophic factor during the medium-conditioning period.

北宗禪硏究 - 頓漸辨證法的 止揚

姜慧? 東國大學校 釋林會 1976 석림 Vol.10 No.-

Clq-coated ELISA법을 이용한 정맥용 면역글로불린제제의 항보체성 측정

강혜나(Hye Na Kang),김순남(Soon Nam Kim),신광훈(Kwang Hoon Shin),허숙진(Sook Jin Hur) 大韓藥學會 2001 약학회지 Vol.45 No.6

The quality of an intravenous immunoglobulin preparation (IVIG) is reflected by the degree of nonspecific activation of complements, the so-called anticomplementary activity (ACA). ACA of aggregates in IVIG was investigated using method by the European Pharmacopoeia and Clq-coated microtiter enzyme-linked immunosorbent assay (ELISA). Both the EP method and the ELISA method showed a dose response curve with the amount of complements bound increasing with the percentage content of aggiegates in immunoglobulin standard. The correlation between the two tests was good (r=0.96, r=0.99). However the correlation was not found when the ACA (EP method) of IVIG product was compared with its aggregate percentage. These results emphasize that the method of aggregate formation affects ACA and that estimation of the percentage distribution of aggregates by HPLC may not reflect ACA. In analysing IVIG product for Clq binding activity test with the ELISA, the result by using Protein A-HRP correlated with aggregate percentage (r=0.84). But the correlation decreased (r=0.48) when the result used Protein A-AP(having poorer sensitivity than HRP) was compared with aggregate percentage. As a result, some variation between the two methods, due to differences in assay principles, is to be expected. However ELISA technique has the advantage in that it is easier to perform, more precise and less subject to reagent variability and is the more suitable screening method than HPLC analysis.

건축유산의 디지털 기술 활용에 관한 국제 연구동향 분석: CIPA2023 국제심포지엄 사례를 중심으로

강혜리,이종욱 한국건축역사학회 2023 건축역사연구 Vol.32 No.6

본 논문에서는 ICOMOS 산하 국제학술위원회에서 개최하는 CIPA 국제심포지엄 사례를 다루고 있다. 해당 심포지엄에서 발표된 연구들을 데이터 획득, 데이터 관리, 데이터 공유 및 체험과 각각의 세부 카테고리로 분류하였고, 이를 통해 디지털 기술을 활용한 건축유산 연구사례를 분석하였다. CIPA2023 국제심포지엄의 학술논문 43건을 연구 주제에 따라 범주화하여 디지털 건축 분야에서의 연구 동향을 분석함으로써 한국건축유산의 디지털 데이터 획득, 관리, 공유 및 체험 분야의 향후 연구 방향을 제시하였다. 결과적으로 각 연구들에서 설계된 알고리즘이나 워크플로우, 방법론 등은 참고 및 보완하여 한국건축유산의 특성과 그 데이터의 특성에 적합한 방법론을 활용할 필요가 있다. Based on my attendance at the CIPA International Symposium(CIPA2023) organized by the International Scientific Committee on Heritage Documentation(ICOMOS), this paper explored research cases applying digital technologies, including BIM, to architectural heritage. The researches presented at this symposium were categorized into specific areas: data acquisition, data management, data sharing&experience. Through this classification, an analysis of research cases in architectural heritage utilizing digital technology was conducted. By categorizing the 43 academic papers from the CIPA2023 based on research themes, trends in the digital architecture field were analyzed, providing insights into future research directions for the digital acquisition, management, sharing, and experiential aspects of Korean architectural heritage. In conclusion, it is deemed necessary to reference and supplement the methodologies, including algorithms, workflows, and approaches developed in each study, to effectively apply methodologies suitable for the characteristics of Korean architectural heritage and its data.