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초기 계대 인간 배아줄기세포의 해동 후 효율적인 배양 방법
백진아,김희선,설혜원,서진,정주원,윤보애,박용빈,오선경,구승엽,김석현,최영민,문신용,Baek, Jin-Ah,Kim, Hee-Sun,Seol, Hye-Won,Seo, Jin,Jung, Ju-Won,Yoon, Bo-Ae,Park, Yong-Bin,Oh, Sun-Kyung,Ku, Seung-Yup,Kim, Seok-Hyun,Choi, Young-Min,Moon, 대한생식의학회 2009 Clinical and Experimental Reproductive Medicine Vol.36 No.4
목 적: 인간 배아줄기세포 (human embryonic stem cells; hESCs)는 미분화 상태로 무한 증식할 수 있는 자가 증식(self-renewal) 능력과 인체의 모든 세포로 분화할 수 있는 전분화능 (pluripotency)의 특징을 가진 세포로, 손상된 세포를 건강한 세포로 대체하고자 하는 세포치료 (cell therapy) 연구에 활용하기 위한 세포 공급원 (cell source)으로 제시되고 있다. 그러나 인간 배아줄기세포는 확립된 초기에 세포를 안정적으로 배양하고 유지하는 과정이 쉽지 않으며, 특히 동결보존되어 있던 세포를 해동한 후 배양할 때 자연 발생적 분화가 높기 때문에 세포주의 유지에 많은 어려움이 따른다. 본 연구에서는 동결보존되어 있던 초기 계대의 인간 배아줄기세포를 해동하여 다시 배양할 때 자연 발생적 분화 부분을 기계적 분리 방법으로 제거하여 미분화 상태의 세포를 보다 빠르게 확보하기 위한 효율적인 방법에 대해 알아보고자 하였다. 연구방법: 인간 배아줄기세포를 계대 배양한지 4일이 되는 날, 50% 이상의 자연 발생적 분화가 나타난 세포군에서 분화된 부분만을 절개용 유리 피펫 (drawn-out dissecting pasture pipette)을 사용하여 기계적인 방법으로 제거하였다. 이후 지속적으로 배양액을 교환해 주며 세포군의 제거된 부분을 7일째 되는 날까지 관찰하였다. 결 과: 기계적 분리 방법을 사용하여 인간 배아줄기세포의 자연 발생적인 분화 부분을 제거한 빈 공간에 미분화상태의 인간 배아줄기세포가 분열하여 채워지는 것을 관찰하였다. 또한, 이 실험 방법을 연속 두 번 적용하여 배양했을 때 미분화 세포로 회복되는 세포군의 비율이 조금 더 높아지는 것을 확인할 수 있었다. 결 론: 동결되어 있던 초기 계대 인간 배아줄기세포의 해동 후, 자연 발생적 분화에 의해 미분화 상태를 유지하는 세포의 수가 적어 계대를 유지 하기가 어려울 때 이와 같은 기계적 분리 방법을 사용하여 자연 발생적 분화 부분을 제거한 후 배양을 지속하는 것이 단기간 내에 미분화 상태를 유지하는 인간 배아줄기세포의 양적 확보를 위한 효율적인 방법이라고 사료된다. Objective: Human embryonic stem cells (hESCs) have the capacity to differentiate into all of the cell types and therefore hold promise for cell therapeutic applications. In order to utilize this important potential of hESCs, enhancement of currently used technologies for handling and manipulating the cells is required. The cryopreservation of hESC colonies was successfully performed using the vitrification and slow freezing-rapid thawing method. However, most of the hESC colonies were showed extremely spontaneous differentiation after freezing and thawing. In this study, we were performed to rapidly collect of early passage hESCs, which was thawed and had high rate of spontaneously differentiation of SNUhES11 cell line. Methods: Four days after plating, partially spontaneously differentiated parts of hESC colony were cut off using finely drawn-out dissecting pipette, which is mechanical separation method. Results: After separating of spontaneously differentiated cells, we observed that removed parts were recovered by undifferentiated cells. Furthermore, mechanical separation method was more efficient for hESCs expansion after thawing when we repeated this method. The recovery rate after removing differentiated parts of hESC colonies were 55.0%, 74.5%, and 71.1% when we have applied this method to three passages. Conclusion: Mechanical separation method is highly effective for rapidly collecting and large volumes of undifferentiated cells after thawing of cryopreserved early passage hESCs.
백진아 ( Jin Ah Baek ),최영민 ( Young Min Choi ),문신용 ( Shin Yong Moon ) 서울대학교 인구의학연구소 2011 人口醫學硏究論集 Vol.24 No.-
The pluripotency and self-renewal of human embryonic stem cells (hESCs) provide appropriate systems for the development of cell replacement therapies. Recent studies have converged on the finding transcription factors, such as Oct4, Sox2, and Nanog, which are served as master regulators in the maintenance of undifferentiated hESC or in early stages of differentiated hESCs. However, downstream targets and signals of these transcription factors for hESC maintenance and regulation are not well characterized. In this study, we summarize the current novel pluripotency markers of hESCs, L1TD1, Banf1, E1B-AP5, POLR3G and L1CAM, that were published in 2011. The identification of novel hESC markers is essential for understanding hESC pluripotency and the mechanisms involved in hESC differentiation and self-renewal.
원용재(Yong-Jae Won),양창인(Chang-Ihn Yang),조영찬(Young-Chan Cho),최임수(Im-Soo Choi),김명기(Myeong-Ki Kim),정응기(Eung-Gi Jeong),이점호(Jeom-Ho Lee),이상복(Sang-Bok Lee),백만기(Man-Kee Baek),홍하철(Ha-Cheol Hong),안억근(Eok-Keun Ah 한국육종학회 2016 한국육종학회지 Vol.48 No.3
The Misomi is a rice variety derived from a cross between ‘Sobi’ which has adaptability of low fertilizer cultivation and SR21110 (Ilpum/Jinbu19) by the rice breeding team at National Institute of Crop Science, RDA in 2013. The heading date of ‘Misomi’ is August 14 and six days later than check variety, ‘Hwaseong’. It has 83 cm of culm length and 121 spikelets per panicle. It showed weaker cold tolerance than that of ‘Hwaseong’ during exposed to cold stress. ‘Misomi’ showed resistance to blast disease, K1, K2 and K3 race of bacterial blight and stripe virus, but susceptibility to K3a race, other viruses and planthoppers. The milled rice of this variety exhibits translucent, clear non-glutinous endosperm and short grain shape. It has low protein content (6.2%) and similar amylose content (19.4%) to that of ‘Hwaseong’. ‘Misomi’ showed better palatability index of cooked rice than that of ‘Hwaseong’. Particulary, ‘Misomi’ showed excellent processing adaptability than ‘Chucheong’ and ‘Hwaseong’ on palatability index of cold and aseptic cooked rice. Its milled rice recovery rate is similar to those of ‘Hwaseong’. However, whole grain rate of milled rice is higher than that of ‘Hwaseong’. Misomi has 5.68 MT/ha of milled rice productivity in ordinary cultivation, which was 12% higher than that of ‘Hwaseong’. Misomi could be adaptable to the middle plain area and Midwest coast area in Korea (Grant No. 5635).
통합적 농촌경관 평가모델 개발 및 적용 -전북 고창선동권역을 대상으로-
반영운 ( Yong Un Ban ),이영훈 ( Yong Hoon Lee ),김민아 ( Min Ah Kim ),최나래 ( Na Rae Choi ),백종인 ( Jong In Baek ) 한국농촌계획학회 2014 농촌계획 Vol.20 No.4
This study has intended to build a rural landscape evaluation model based on an integrated landscape assessment paradigm of rural region using an additive integration index method and applied the model to the Seondong Region of Gochang-gun, Jeollabuk-do, Korea. To reach this goal, this study developed a model to calculate Integrated Landscape Assessment Index. The model has employed the Objective Landscape Index, the Subjective Landscape Index, and the weighted values, and was applied to the Seondong region. This study has found the following results: 1) forests and water spaces were assessed with relatively better visual preferences and better landscape ecosystem; 2) the historic cultural area and scenic agriculture as well as general farm land were assessed with moderate ratings; and, 3) the villages included in development plan, their adjacent arable farming land, and the village watercourses were forming relatively poorer landscape.
반영운 ( Yong Un Ban ),이태호 ( Tae Ho Lee ),백종인 ( Jong In Baek ),김민아 ( Min Ah Kim ) 한국도시지리학회 2012 한국도시지리학회지 Vol.15 No.2
본 연구의 목적은 농촌경관의 주관적 평가를 위해 경관영역 및 공간유형별 대표경관형용사를 선정하는 것이다. 이 연구는 다음과 같이 3단계로 구성된다. 첫째, 농촌경관과 경관 형용사의 개념에 대한 정의를 위해 관련 문헌을 고찰하고, 그 정의에 기초하여 경관형용사를 선정하였다. 둘째, 농촌 경관 평가에 적합한 경관형용사 선정을 위해 1차 설문조사를 실시하였다. 셋째, 각각의 공간 유형에 적합한 대표 경관형용사 선정을 위해 2차 설문조사를 실시하였다. 그 결과로, 본 연구는 경관영역 및 공간유형별로 12개의 대표 경관형용사를 선정하였다. This study has intented to select representative landscape adjectives in accordance with both landscape units and space types for subjective evaluation of rural landscape. This study was composed of the following stages: 1) to define the concept of the rural landscape and landscape adjectives through reviewing related literatures, and to list the landscape adjectives based on the definitions, 2) to execute primary survey to select landscape adjectives, best fit to evaluate the rural landscape, 3) to execute subsequent survey to select representative landscape adjectives, adequate to each space type. As a result, this study has selected a total of 12 representative landscape adjectives according to both landscape units and space types.
정상과 비정상핵형을 가진 인간 배아줄기세포의 비교 분석
설혜원 ( Hye Won Seol ),백진아 ( Jin Ah Baek ),정주원 ( Ju Won Jung ),윤보애 ( Bo Ae Yoon ),김희선 ( Hee Sun Kim ),오선경 ( Sun Kyung Oh ),구승엽 ( Seung Yup Ku ),김석현 ( Seok Hyun Kim ),문신용 ( Shin Yong Moon ),최영민 ( Young M 서울대학교 인구의학연구소 2011 人口醫學硏究論集 Vol.24 No.-
Human embryonic stem cells (hESCs) are a useful source for biological research fields and applications in regenerative medicine. The maintenance and distribution of the undifferentiated hESCs of normal karyotypes are very important. The changes of colony morphology were observed during the culturing of hESCs, and chromosomal analysis was performed in this study. Abnormal karyotypes were identified in the cells which have the changes of the colony morphology. Attachment ratio and the spontaneous differentiation ratio of abnormal cells were compared to those of normal cells. High cell attachment ratio and low spontaneous differentiation ratio were confirmed in karyotypically abnormal cells. HESCs showed chromosomal anomaly if the colony morphology changed, and high attachment ratio and low spontaneous differentiation ratio have been observed during the cell culture. Our results suggest that the morphology of the colonies would play a ro1e as a criterion for chromosomal anomaly.