세포 배양 시 마이코플라즈마의 검출 및 제거 방법에 대한 연구 동향

윤보애 ( Bo Ae Yoon ),오선경 ( Sun Kyung Oh ),최영민 ( Young Min Choi ) 서울대학교 인구의학연구소 2011 人口醫學硏究論集 Vol.24 No.-

Mycoplasma contamination may have multiple effects on cultured cells and can also have significant influences on the results of scientific studies. Therefore, the elimination of mycoplasma contamination from cell cultures is necessary. This review provides an overview of the following research trends: (1) mycoplasma contamination in cell cultures; (2) various techniques available for the detection of mycoplasma; (3) four method types available for the elimination of mycoplasma We will discuss the problems of mycoplasma contamination in cell cultures and present a concise overview on the biology of mycoplasma with suggestions on efficient methods for its detection and elimination, and proposing steps to be taken to prevent the problems.

인간 배아줄기세포의 효율적인 동결방법 연구 동향

윤보애 ( Bo Ae Yoon ),설혜원 ( Hye Won Seal ),최영민 ( Young Min Choi ) 서울대학교 인구의학연구소 2012 人口醫學硏究論集 Vol.25 No.-

Human embryonic stem cells (hESCs) are one of the most interesting cell types for tissue engineering, cell therapy, basic scientific research, and drug screening. Fast advancement in these areas requires the availability of large amounts of safe and well-characterized hESCs from hESC banks. The sensitivity of hESCs to cryopreservation, however, has severely limited the utility of these cells. It was observed that the characteristics of hESCs changed during long-term culture. Therefore, it is crucial to preserve early-passage cells by effective freezing and thawing methods. This paper reviews the current state of cryopreservation as it pertains to hESCs. We focused on the research of cryopreservarion method of hESCs.

인간 배아줄기세포 배양을 위한 영양세포의 최적밀도에 관한 연구

윤보애 ( Bo Ae Yoon ),설혜원 ( Hyun Won Seol ),백진아 ( Jin Ah Baek ),서진 ( Jin Seo ),정주원 ( Ju Won Jung ),김희선 ( Hee Sun Kim ),오선경 ( Sun Kyung Oh ),문신용 ( Shin Yong Moon ),최영민 ( Young Min Choi ) 서울大學校 醫學硏究院 人口醫學硏究所 2009 人口醫學硏究論集 Vol.22 No.-

인간 배아줄기세포의 feeder-free 배양 환경에 사용된 다양한 기질 고찰

윤보애 ( Bo Ae Yoon ),문신용 ( Shin Yong Moon ),최영민 ( Young Min Choi ) 서울대학교 인구의학연구소 2010 人口醫學硏究論集 Vol.23 No.-

Human embryonic stem cells(hESCs) have been derived and cultured in various condition. Culture of undifferentiated hESCs traditionally requires mouse fibroblasts as a feeder cell layer. The hESC culture using mouse fibroblasts makes clinical application of the hESCs difficult because that the mouse fibroblasts could give a risk of contamination with xeno-pathogens. Therefore, replacing feeder cells with defined human extracellular matrix(ECM) components or synthetic biomaterials would be ideal for the maintenance of clinical-grade hESCs. In this review, we discuss with recent advances in the use and development of substrates suitable for feeder-free culture condition.

미국과 영국의 줄기세포은행 시스템 비교

윤보애 ( Bo Ae Yoon ),오선경 ( Sun Kyung Oh ),최영민 ( Young Min Choi ) 서울대학교 인구의학연구소 2013 人口醫學硏究論集 Vol.26 No.-

Human embryonic stem cells (hRSCs) have unlimited potentials of proliferation and differentiation and these properties of hESCs make them considered as most interesting cell types for tissue engineering, cell therapy, and drug screening. Stem cell bank mainly do quality control of stem cell lines including best practice in culture, preservation and provide information for the choice of proper embryonic stem cell (ESC) line to proceed further studies. Currently, USA and UK are leading counties for stem cell research and banking. This paper reviews, the current state of the USA and the UK stem cell bank and compared the differences between two banks.

CLN6 유전자 변이에 의한 진행근간대뇌전증 1예

이현경(Hyun Gyung Lee),윤보애(Bo Ae Yoon),김영옥(Young Ok Kim),김명규(Myeong Kyu Kim),우영종(Young Jong Woo) 대한소아신경학회 2018 대한소아신경학회지 Vol.26 No.2

NCLs는 선천성 대사 이상 질환의 하나로, 리소좀에 자가 형광물질인 lipopigment가 축적되어 발생하는 신경퇴행성 질환이다. 2세에서 4세 사이에 발병하는 전형적인 영아 후기형 NCL에 비해 더 늦은 나이인 3세에서 8세 사이에 경련과 점진적인 시력 소실이 시작되는NCLs를 변이 영아 후기형 NCLs (Variant forms of late infantile NCLs,vLINCLs)이라고 하며, 이는 CLN6 에 의해 생긴다고 알려져 있다. 저자들은 진행성 근간대 뇌전증으로 진단된 여아에서 whole-exome sequencing을 통해서 CLN6 유전자 (NM_017882.2; NP_060352.1)의 c.296A>G(p.Lys99Arg)와 c.307C>T(p.Arg103Trp) 이형접합체 변이들을 발견하였고 이를 Sanger sequencing으로 확인하였다. 환아는 7세 이후에 여러 형태의 경련을 보였고, 저명한 광과민성 근간대 경련, 진행성 운동 및 언어능력 퇴행, 추체로 및 추체외로 징후, 뇌자기공명 검사에서 진행성 뇌위축 등을 vLINCLs의 전형적인 특징을 보였다. 본 증례는 한국인에서 첫번째로 보고되는 CLN6 유전자 변이에 의한 vLINCLs보고이며, 일본에서 보고된 증례와 같이 청소년기에도 시력 소실을 보이지 않았다. Neuronal ceroid lipofuscinoses (NCLs) are inherited neurodegenerative disorders, which are caused by the accumulation of lipopigment in lysosomes. Variant forms of late infantile NCLs (vLINCLs) characterized by a later onset of seizures and visual impairment (3–8 years) than in the classic form (2–4 years) are caused by mutations of the gene encoding ceroid lipofuscinosis neuronal protein 6 (CLN6). In a girl with progressive myoclonus epilepsy, we found heterozygous variants of CLN6 (NM_ 017882.2; NP_060352.1): c.296A>G (p.Lys99Arg) and c.307C>T (p.Arg103Trp). They were identified with whole-exome sequencing and verified with Sanger sequencing. At 7 years and 9 months, our patient had developed multiple types of seizures, prominent myoclonus with photosensitivity, regression in motor and language skills, pyramidal and extrapyramidal signs, and brain atrophy in brain images, all of which were progressive and were compatible with vLINCLs. However, this first Korean report shows no visual impairment, which resembles the previously reported Japanese case.

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.

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.