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최인경,송해범 대구대학교 생명과학연구소 2004 생명과학연구 Vol.3 No.1
Abstract : The cryopreservation of embryos has become a powerful tool in assisted reproduction in several mammalian species. Embryos are cryopreserved by slow freezing or by vitrification. However, consistently high survival has not been obtained in most oocytes and in some embryos. The main reasons for the low survival would be sensitivity to low temperatures, which leads to chilling injury, and low permeability of the cell membrane, which leads to the formation of intracellular ice. As a strategy aiming to overcome these injuries, modified vitrification methods have been devised in which the cooling and warming rate is markedly incresed by minimizing the column of the solution and the container. Furthermore, porcine oocytes and embryos are influenced just by chilling to below 15℃.
배양액의 에너지원 조성이 생쥐 초기배 발달에 미치는 효과에 관한 연구
이종범(Jong Bum Lee),김주환(Ju Hwan Kim),고지환(Jee Hwan Ko),오영균(Young Kun Oh),손성경(Song Kyong Son),서영석(Young Seok Seo),노흥태(Heung Tae Noh),강길전(Kil Chun Kang),송해범(Hai Bum Song),이기환(Ki Hwan Lee) 대한산부인과학회 2002 Obstetrics & Gynecology Science Vol.45 No.3
N/A Objective: The objective of this study was to examine the effect on development of mouse preimplantation embryos in culture media with different composition of energy sources in vitro culture. Methods: Two hundred and seventy one two-cell embryos were cultured in four different culture system for 96 hours. Group I (n=61)was cultured in DMEM-G (DMEM with glutamine) only, group II (n=64)was cultured in DMEM-GGP (DMEM with glutamine, glucose and pyruvate) only, group III(n=72) was cultured for 48hoursinDMEM-G and then transferred to DMEM-GGP and group IV (n=74)was cultured for 48 hours in DMEM-GGP and then transferred to DMEM-G. Development of embryos in each group was observed every 24 hours. Results: After 24 hours, the rate of development ≥3-cell was significantly higher in group II(87.5%) and IV(86.5%)compared with group I(59.0%)and III(62.5%).After 48 hours, the rate of development into ≥morula stage was significantly higher in Group II(79.7%)and IV(86.5%)compared with group I(34.4%) and III(37.5%).After 72 hours, the rate of development into blastocyst was significantly higher in group IV(74.3%)compared with group I (49.2%)and III (45.8%).After 96 hours, the rate of development into ≥expanded blastocyst was significantly higher in group IV(70.3%)compared with group I (32.8%), II ( 53.1%), and group III (40.3%) Conclusion: Mouse preimplantation embryos development was the most effective in culture system with DMEM-GGP for 48 hours and then transferred to DMEM-G.
알지네이트가 가교된 소장점막하조직 스폰지의 생체적합성 평가
송인범 ( In Bum Song ),이민숙 ( Min Suk Lee ),김문석 ( Moon Suk Kim ),강길선 ( Gil Son Khang ),이해방 ( Hai Bang Lee ) 한국조직공학과 재생의학회 2006 조직공학과 재생의학 Vol.3 No.3
Porcine small intestine submucosa(SIS), mainly composed of collagen and glycosaminoglycan, has been widely used as a material for organ and tissue reconstruction without immuno-rejection responses. Chemicallycrosslinked SIS sponges were prepared and characterized for bio-interactive wound dressings and tissue engineered scaffolds. SIS powder were reacted in aqueous solution of 3% acetic acid and 0.1% pepsin for 48 hrs and the prepared SIS solution was poured into mold and fabricated by freeze-drying method. The crosslinking reaction was performed using 1-ethyl-(3-3-dimethyl aminopropyl) carbodiimide hydrochloride(EDC) solution(distilled water(D.W.) : ethanol = 5 : 95) of 50, 100 and 200 mM concentration for 24 hrs. Then 1 wt% alginate solution was pass through the SIS sponge and lyophilzed. 102 mM calcium chloride(CaCl2) was used as crosslinking agent. The prepared sponges were characterized by scanning electron microscopy(SEM). The biocompatibility of NIH/3T3 fibroblast with these SIS sponges was found to be acceptable at a cell density of 2×104 cell/cm2. We investigated biodegradation of SIS sponge, MTT assay and H&E stain. From the MTT assay results, the biocompatibility of the crosslinked SIS sponge with NIH/3T3 fibroblast was confirmed. It was confirmed that SIS consists of organic components like tissue and showed inter-connective pores for adhesion and growth of cell. As a result of this study, we suggest that crosslinked alginate-SIS sponges fulfil many critical elements desirable in the application of wound dressing and tissue regeneration material.
Cryopreservation and Cryobiology of Mammalian Oocytes
Kasai, Magosaburo,Song, Hai-Bum 대구대학교 생명과학연구소 2003 생명과학연구 Vol.1 No.3
The cryopreservation of embryos has become a powerful tool in assisted reproduction in several mammalian species and a more important technology in bioscience, agriculture and medicine. Embryos are cryopreserved by slow freezing or by vitrification. However, consistently high survival has not been obtained in most oocytes and in some embryos. The main reasons for the low survival would be sensitivity to low temperatures, which leads to chilling injury, and low permeability of the cell membrane, which leads to the formation of intracellular ice. As a stratefy aiming to overcome these injuries, modified vitrification methods have been devised in which the cooling and warming rate is markedly increased by minimizing the volume of the solution and the container. The modified methods use electron microscope grids, open-pulled straws, cryoloops, or container-less microdrops. Ultrarapid vitrification is a promising approach to overcoming the problems of the chilling sensitivity and lower permeability of oocytes/embryos. However, to make this method more reliable, defining the optimal combinations of cooling rate and the composition of the vitrification solution for each type of oocyte/embryo will be necessary. Refinement of the procedures for practical use, including labeling and stable preservation, would also be important.