필수수기검사와 문제해결 중심의 임상병리학 임상교육

양윤선(Yoon Sun Yang),조한익(Han Ik Cho),박성섭(Sung Sup Park),김진규(Jin Q Kim),김상인(Sang In Kim) 한국의학교육학회 1992 Korean journal of medical education Vol.4 No.2

To improve practical skills and problem-solving ablihty of the medical students, authrs introduced unique teaching program composed of 10 laboratory tests, essential for primary care physicians and ar ound 200 problem-solving and actual questionaires which can be met in situations of the clinical setting. The program was applied to the two-week clerkship for the senior class of Seoul National University College of Medicine in 1992. The problem-solving questionaires were revised three times in 1992, according to the results of analysis for new questionaires made by teachers and students. Around 5% of practical problem-oriented questionaires were made by students themselves. Favorable responses from the students were 99.1% for overall program and 96.1% for problem-solving questionaires. The long-term impact of this clerkship should be re-evaluated when students educated by this program become primary care physicians including interns or residents.

인간 제대혈 유래 간엽줄기세포의 연골세포 분화

정미현(Mee Hyun Jung),양성은(Sung-Eun Yang),진혜진(Hae Jin Jin),이만경(Man Kyoung Lee),송호선(Ho Sun Song),양정윤(Jung Yoon Yang),양윤선(Yoon Sun Yang),하철원(Chul-Won Ha) 대한정형외과학회 2004 대한정형외과학회지 Vol.39 No.6

목적: 인간 제대혈로부터 간엽성 섬유세포양 세포를 분리, 배양함으로써 제대혈 내의 순환성 간엽줄기세포의 존재여부를 확인하고, 이의 연골세포 분화능을 증명하고자 하였다. 대상 및 방법: 총 50 유니트의 제대혈로부터 분리된 유착성 세포군의 세포형태 및 면역표현형을 분석하였다. BMP-6을 첨가하였거나 또는 첨가하지 않은 연골분화배양액을 이용하여 펠렛 배양의 형태로 연골분화를 유도 배양하였다. 연골세포로의 분화를 관찰하기 위해 역전사 중합효소연쇄반응을 수행하였으며, Safranin-O 염색과 제2형 콜라겐에 관한 면역염색을 시행하였다. 결과: 제대혈 단핵세포로부터 균질성의 섬유세포양 형태의 유착성 세포군이 배양되었다. 이 세포군은 간엽줄기세포 관련 항원 양성, 조혈모세포항원 음성, 조직적합항원 음성, 내피세포 및 파골세포 관련 항원 음성이었다. 연골분화 펠렛 배양 시 BMP-6을 첨가한 군에서 펠렛의 크기 및 연골세포관련 인자의 발현이 증가하였고, Safranin-O 염색과 제2형 콜라겐의 면역염색도 강양성이었다. 결론: 제대혈 내에 순환성 비조혈성 연골형성 간엽줄기세포가 존재함을 확인하였고, 연골분화 배지에 BMP-6을 첨가함으로써 제대혈 유래 비조혈성 간엽줄기세포의 시험관 내 연골분화의 효율을 향상시킬 수 있었다. Purpose: The aim of this study was to demonstrate the existence of circulating mesenchymal stem cells (MSC) in the human umbilical cord blood (hUCB) and to evaluate the chondrogenic differentiation potential of hUCB-derived MSC in vitro. Materials and Methods: Fifty hUCB harvests were cultured in media supplemented with 10% fetal bovine serum. The adherent fibroblast-like cells were characterized by immunophenotyping and induced to differentiate into chondrocytes in the pellet culture with and without BMP-6. This study performed RTPCR of the chondrogenic markers, Safranin-O stain and type Ⅱ collagen immunohistochemical stain. Results: The mononuclear cells isolated from hUCB formed adherent colonies with an attached well-spread fibroblast-like morphology. The cells positively expressed the MSC-related antigens, but did not express the hematopoietic, HLA-DR, endothelial, or osteoclast antigens and could be induced to differentiate into chondrocytes under proper stimulation. BMP-6 increased the size of the pellet and the mRNA levels for aggrecan, type Ⅱ collagen and type Ⅸ collagen and enhanced the levels of proteoglycan synthesis during chondrogenic differentiation. Conclusion: The homogenous fibroblast-like cells developed in cultures from hUCB with chondrogenic differentiation potential were considered to be MSC. Furthermore, it was found that BMP-6 enhanced chondrogenic differentiation of the hUCB-derived MSC in the pellet culture.

Isolation, Characterization and Tri-lineage Differentiation of Mesenchymal Stem Cells from Human Umbilical Cord Blood

Sung-Eun Yang(양성은),Yoon-Sun Yang(양윤선),Chul-Won Ha(하철원) 대한정형외과학회 2004 대한정형외과학회지 Vol.39 No.5

목적: 저자들은 인간 제대혈(umbilical cord blood)에서 간엽 줄기 세포를 분리 및 확인할 수 있었고, 이들 세포군에 대하여 골, 연골 및 지방세포로의 분화가 가능하였기에 이를 보고하고자 한다. 대상 및 방법: 제대혈(n=411)로부터 단핵세포군을 원심분리 방법을 이용하여 분리한 후 배양하였으며, 5-8회 개대배양 시에 1,000배 이상의 증식을 보인 세포군에 대하여 간엽줄기세포 및 타 세포계열 표면항원 검사를 시행하였다. 분리된 간엽줄기 세포군에 대하여 골, 연골 및 지방세포로의 분화를 시도하였다. 결과: 총 411개의 제대혈 중 95개(23.1%)에서 첫 배양 시 간엽 줄기 세포양 세포군(MSC-like cell population)이 확인되었으며, 5-8회 개대배양 시 1,000배 이상의 증식을 보인 세포군은 9개(2.2%)였다. 이들 세포군은 기존에 알려진 모든 간엽줄기세포 표면항원검사에 양성, 타 세포계열 표면항원 검사에는 모두 음성이었다. 분리된 간엽 줄기 세포군은 골, 연골 및 지방세포로의 분화가 가능하였다. 결론: 본 연구의 결과는 인간 제대혈이 서로 다른 세포 계열로 분화할 수 있는 간엽줄기세포를 포함하고 있다는 사실을 지지하는 것으로 판단된다. 인간 제대혈은 상대적으로 많은 양의 간엽줄기세포를 비교적 용이하게 얻을 수 있다는 장점이 있는 세포 공급원이 될 수 있을 것으로 사료되며, 향후 조직 공학 및 세포 치료를 위한 세포 공급원의 하나가 될 수 있을 것으로 기대한다. Purpose: Human umbilical cord blood (hUCB) is well known for a good source of hematopoietic stem cells (HSCs). However, the presence of mesenchymal stem cells (MSCs) in hUCB is still not well approved by many authors. We hereby report the isolation and characterization of MSCs from hUCB, as well as their differentiation into osteogenic, chondrogenic and adipogenic lineages. Materials and Methods: Mononuclear cells were isolated from each hUCB harvest (n=411) by density gradient centrifugation, and suspended in -minimum essential medium supplemented with 10% fetal bovine serum (FBS). The cell population was expanded by successive sub-cultivation under the same condition. The cell population that showed more than 1,000-fold expansion at fifth to eighth passage was inspected with known surface antigens of MSCs and other cell lineage. The isolated MSCs were cultured in osteogenic, chondrogenic, and adipogenic condition to identify their potential to differentiate into different mesenchymal cell lineage. Results: Ninety five out of 411 hUCB units (23.1%) generated the MSC-like cell population during initial cultivation. Nine cell populations (2.2%) showed more than 1,000-fold expansion capacity at fifth to eighth passage. These cells positively expressed all known MSC-related antigens. They did not express any of myeloid, endothelial, or histo-compatibility antigens. All of the MSCs isolated showed the potential to differentiate into osteogenic, chondrogenic, and adipogenic lineages. Conclusion: Our study supports that hUCB does contain MSCs, which can be differentiated into different cell lineages. We believe hUCB will be a good source of MSCs with the advantage of availability and relative abundance. We think hUCB should not be considered as a medical waste, and it will serve as a good source of cells for tissue engineering and cellular therapy in the future.

한국인 전신성홍반성루푸스 환자에서 HLA-DRB1, DQB1 대립유전자의 연관성 및 항인지질 항체와 항β₂ Glycoprotein I 항체에 관한 연구

이상곤,차훈석,양윤선,Lee, Sang Gon,Cha, Hoon Suk,Yang, Yoon Sun 대한면역학회 2002 Immune Network Vol.2 No.4

Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by diverse clinical manifestations and autoantibody production, which is known to be strongly influenced by genetic factors. Previous studies have revealed the associations of SLE with HLA class II alleles and antiphospholipid antibody system (anticardiolipin antibody (aCL) and anti-${\beta}_2$ glycoprotein I antibody (anti-${\beta}_2$ GPI)). Therefore, we studied the associations of HLA class II alleles with SLE and antiphospholipid antibody system. Methods: The genotyping for HLA-DRB1 and DQB1 alleles were performed in 61 SLE patients and 100 controls by the polymerase chain reaction (PCR)-sequence specific oligonucleotide probe method. ELISA tests for aCL and anti-${\beta}_2$ GPI were performed in 39 of the 61 SLE patients. The results were evaluated statistically by Chi-square test. Results: The frequencies of the HLA-$DRB1^*15$ and $DQB1^*06$ in SLE patients were significantly higher than those in controls. HLA-$DRB1^*12$ was significantly lower in SLE patients than controls. Nine of 39 patients were positive for aCL (IgG) and three were positive for aCL (IgM). One of 39 patients were positive for anti-${\beta}_2$ GPI (IgG) and none of them positive for anti-${\beta}_2$ GPI (IgM). Association of aCL with HLA class II alleles was not observed in our study. Conclusion: According to our results, it was found that HLA-$DRB1^*15$ and $DQB1^*06$ were associated with genetic susceptiblility and $DRB1^*12$ was associated with resistance to SLE in Korean population. No Association of aCL with HLA class II alleles was observed and the positive rate for anti-${\beta}_2$ GPI was very low.

간엽줄기세포를 이용한 자가 면역 질환의 치료

이재권 ( Jae Kwon Lee ),양성은 ( Sung Eun Yang ),양윤선 ( Yoon Sun Yang ),오원일 ( Won Il Oh ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.3

한국인 전신성홍반성루푸스 환자에서 HLA-DRB1, DQB1 대립유전자의 연관성 및 항인지질 항체와 항β₂ Glycoprotein I 항체에 관한 연구

이상곤(Sang Gon Lee),차훈석(Hoon Suk Cha),양윤선(Yoon Sun Yang) 대한면역학회 2002 Immune Network Vol.2 No.4

자궁내막증 환자의 말초혈액 및 복강액에서의 면역 세포 아형과 cytokine 농도의 특성

이제호(Je Ho Lee),김정구(Jung Gu Kim),최두석(Doo Seok Choi),이정원(Jeong Won Lee),윤병구(Byung Koo Yoon),양윤선(Yoon Sun Yang),김대원(Dae Won Kim),김동호(Dong Ho Kim) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.12

N/A Objective: Endometriosis is a common and enigmatic disease affecting the reproductive life and health of women. Although the retrograde menstruation is a well established model for both transplantation and induction theories, the discrepancy between an incidence of retrograde menstruation and a prevalence for endometriosis suggests the possibility that the development and the progression of endometriosis is associated with individual susceptibility such as altered immune function. An impaired immune response may result in a defect in the ability to remove refluxed menstrual debris, thereby increasing the possibility of endometriosis. We carried out the study to elucidate the immunologic alteration in patients with endometriosis. Materials and Methods: Fifty-six patients undergoing pelviscopic surgery or open laparotomy for benign gynecological disease were enrolled in this study. The study groups consisted of group I (normal control patients, N=22), group II (endometriosis stage I and II, N 17), and group III (endometriosis stage III and IV, N=17). Lymphocyte subset including total T cell, helper T cell, suppressor T cell, B cell, helper/suppressor ratio, natural killer (NK) cell, monocyte population and cytokine profile including interleukin (lL)-1, soluble interleukin-2 receptor (slL-2R), IL-2, IL-6, IL-S, monocyte chemoattractant protein (MCP)-1 of peripheral blood and peritoneal fluid were analyzed using flow cytometry and enzyme-linked immunosorbant assay (ELISA) method respectively. Results: Peripheral blood and peritoneal fluid lymphocyte subset were indistinguishable among the 3 groups (p>0.05). And there were no significant difference in peripheral blood and peritoneal fluid cytokine profile among the 3 groups except peripheral blood MCP-1 level. Group III showed higher peripheral blood level of MCP-1 than control patients (p<0.05). Conclusion: In this study, lymphocyte subset and cytokine profile except MCP-1 in peripheral blood and peritoneal fluid from patients with endometriosis did not differ from those of the control group. Immunologic alterations of patients with endometriosis might be resulted not from the changes of the number of lymphocyte subsets and cytokine, but from the modification of functions.

일반총설 : 척추 추간판탈출증(디스크)에 있어서 세포치료제 연구개발의 최근동향

김순희 ( Soon Hee Kim ),고연경 ( Youn Kyung Ko ),하현정 ( Hyun Jung Ha ),윤선중 ( Sun Jung Yoon ),양윤선 ( Yoon Sun Yang ),김문석 ( Moon Suk Kim ),강길선 ( Gilson Khang ),이일우 ( Il Woo Lee ),이종문 ( John M. Rhee ),이해방 ( Hai 한국조직공학과 재생의학회 2007 조직공학과 재생의학 Vol.4 No.3

연령이 골수 간엽줄기세포 분화능력에 미치는 영향

진혜진(Hye Jin Jin),최수진(Soo Jin Choi),배윤경(Yun Kyung Bae),이수연(Soo Yeun Lee),길명옥(Myeong Og Gil),박찬미(Chan Mi Park),왕준호(Joon Ho Wang),오원일(Wonil Oh),양윤선(Yoon Sun Yang),조인호(Inho Jo),김철기(Cheol-Ki Kim),송해룡(Hae-R 대한정형외과학회 2007 대한정형외과학회지 Vol.42 No.6

목적: 인간 골수로부터 간엽줄기세포를 분리 배양하여 기증자의 나이에 따른 분화능력을 비교하였다. 대상 및 방법: 저연령 그룹 (n=16, 12.5±5.8세)와 고연령 그룹 (n=4, 48.5±7.2세)으로 나눠 총 20명의 환자로부터 동의를 얻어 골수를 채취하였고, 채취한 골수로부터 간엽줄기세포를 추출하였으며, 세포 형태와 면역표현형을 분석하였다. 또한, 세포의 노화를 두 그룹에서 계대 배양하여 측정하였다. 골, 연골, 지방세포로의 분화를 관찰하기 위해 역전사 중합효소연쇄반응을 수행하였으며, alkaline phosphatase, von Kossa, safranin O와 oil red O 염색을 시행하였다. 결과: 총 20명의 골수에서 모두 간엽줄기세포를 추출하였고, 저연령 그룹에서 획득한 간엽줄기세포의 수가 고연령 그룹에 비해 약 5배 많았다. 세포의 노화는 두 그룹 모두 10세대 이후에서 나타났으며, 면역표현형도 두 그룹에서 유사한 형태를 보였다. 골, 연골, 지방세포로의 다양한 분화능력을 보였으며, 두 그룹간의 차이는 없었다. 결론: 본 연구의 결과는 골수 기증자의 나이는 골수 간엽줄기세포의 분화능력에 영향을 주지 않음을 보여 주었다. Purpose: The aim of study was to compare the differentiation capacity of mesenchymal stem cells (MSCs) obtained from human bone marrow (BM) according to the age of the donors. Materials and Methods: MSCs were isolated from the BM of young (n=16, 12.5±5.8 years) and elder (n=4, 48.5±7.2 years) patients with the consent of them. We analyzed the cell morphology and the cell surface markers of the MSCs. In addition, we assessed the cell senescence with serial cultures from both age groups. Cell pluripotentiality was analyzed by osteogenic, chondrogenic, and adipogenic induction media. We performed RT-PCR, a measurement of expression of alkaline phosphatase, and staining with von Kossa, safranin O, and oil red O stain. Results: All of the MSC samples tested, irrespective of the age of the donors, MSCs were all successfully isolated from twenty bone marrows. However, the number of cells of from the young donors was five times greater than that of the elderly donors. Senescence was observed over 10 passages in both age groups. The immunophenotypes of both age groups showed similar patterns. MSCs obtained from young and older donors showed the potential to differentiate into osteogenic, chondrogenic, and adipogenic lineages with no difference for both age groups. Conclusion: Our study supports that age does not influence the pluripotential capacity of human BM derived MSCs.

HLA-DR Genotyping 키트 "INNO-LiPA HLA DRB"의 평가

양윤선,김대원 대한임상병리학회 1996 대한임상병리학회지 Vol.16 No.2