• 제목/요약/키워드: differentiation therapy

검색결과 347건 처리시간 0.03초

사람의 제대 및 양막유래 줄기세포의 간세포로의 분화 (Hepatogenic Potential of Umbilical Cord Derived-Stem Cells and Human Amnion Derived-Stem Cells)

  • 김지영;이윤정;박세아;강현미;김경식;조동제;김해권
    • Clinical and Experimental Reproductive Medicine
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    • 제35권4호
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    • pp.247-265
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    • 2008
  • 목 적: 사람의 제대유래 줄기세포 (HUC)와 양막유래 줄기세포 (HAM)를 간유사세포로 분화 시키고자 하였다. 연구방법: 산모의 동의아래 만삭 정상 산모로부터 양막 및 제대를 얻고 줄기세포를 분리, 배양하였다. 분리된 세포의 줄기세포로서의 특성을 규명하기 위하여 RT-PCR, 세포면역화학 분석 그리고 중배엽성세포로의 분화능 실험을 하였다. 또한 이들 줄기세포를 분화 배양액에서 간유사세포로 분화 유도 후, 간세포 특이 유전자 발현, 알부민 ELISA, 알부민에 대한 면역블로팅 및 세포면역화학염색 그리고 PAS 염색을 시행하였다. 결 과: 사람의 양막 및 제대로부터 세포를 분리하여 줄기세포로서의 특성을 조사하고, 이들 세포의 지방세포, 연골세포, 골아세포로의 분화능력을 확인하였다. 동일한 분화 배양액에서 HUC과 HAM을 간유사세포로 분화시킨 결과 HUC이 모든 조건에서 HAM보다 알부민과 요소를 더 많이 합성 분비하였다. 간유사세포로의 분화능력이 더 좋은 것으로 나타난 HUC을 가지고 최적의 분화를 유도한 결과 $1.2{\pm}0.8\;{\mu}g/mL$의 알부민을 합성 분비하였고, $8.9{\pm}1.2\;mg/dL$의 요소를 합성 분비하였다. 결 론: HUC과 HAM 모두 알부민을 분비하는 간유사세포로 분화할 수 있었으며, HUC이 더 잘 유도되는 것으로 관찰되었다. 이러한 결과로써 HUC과 HAM을 간질환 환자의 간세포 이식 치료에 필요한 세포치료제로써 이용할 수 있을 것으로 생각된다.

창이자 추출물이 아토피 피부염 유발 생쥐의 비장 세포 Th17의 세포분화 억제에 따른 아토피 피부 상태에 미치는 영향 (Studies of Xanthium strumarium Extract Suppressing Th17-cell Differentiation and Anti-dermatitic Effect in BMAC-induced Atopy Dermatitis of NC/Nga Mice)

  • 김금란;최태부
    • KSBB Journal
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    • 제24권4호
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    • pp.383-392
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    • 2009
  • 아토피피부염 (AD)은 천식, 음식 알레르기, 비염 같은 전신아토피질환을 동반하는 만성재발성 피부염증질환이다. 아토피피부염과 관련된 IL-17의 임상적 역할은 다양한 조건에서 보고되고 있으며, 또한 건선 피부 상태에 깊숙이 관여하고 있다. IL-17은 각질세포 (keratinocytes)에서 과잉으로 생산되며, 아토피피부염의 말초임파구에서도 다량 생성됨을 세포내염색을 통하여 확인된 바 있다. 본 연구에서는 창이자 추출물 (XS-E와 XS-FL)이 NC/Nga 생쥐의 $CD4^+$ T 세포에서 유도된 Th17 세포의 분화억제 및 IL-17의 생산량 감소 효과에 대한 실험을 하였다. 그 결과 XS-E와 XS-FL을 처리한 섬유아세포에서 세포독성은 나타나지 않았고, 4일간 XS-E와 XS-FL에 동시배양 한 $CD4^+$ T 세포의 IL-17 생산량을 FACS로 분석한 결과 $100\;{\mu}g$/mL XS-E 처리군의 IL-17 생산량은 32.3%로 대조군에 비하여 2배 이상의 감소를 나타내었으며, $20\;{\mu}g$/mL XS-30% AFL (acetone XS-FL) 처리군의 Th17 세포는 19.6%으로 대조군에 비하여 3.5배 억제되었다. 또한 real-time PCR을 이용하여 IL-17A와 IL-22 mRNA의 유전자 발현량을 비교 분석한 결과, IL-17A와 IL-22 mRNA의 유전자발현의 RQ값은 XS-E와 XS-30% AFL를 처리한 실험군이 대조군에 비하여 유의성 있는 감소를 나타내었다(p < 0.01, p < 0.001). ELISA로 측정한 IL-17A 생산량은 XS-E와 XS-30% AFL를 처리한 실험군이 대조군에 비하여 현저히 감소 (p < 0.05, p < 0.001)하였다. Th17 세포의 증식을 알아보기 위하여, rIL-6와 TGF-$\beta$로 분화시킨 Th17 세포를 CFSE로 표지한 후 rIL-23 처리를 하여 4일간 배양하여 증식을 유도시켰다. 대조군의 Th17 세포 분열은4일 동안 4번에 걸쳐 비슷한 세포수의 증식이 일어나는 것을 CFSE를 통하여 확인하였고, XS-30% AFL 처리군은 CFSE의 형광 분포가 점점 감소하여 Th17 세포의 증식이 억제됨을 알 수 있었다.

국소 진행된 간암의 방사선 온열치료성적 (The Clinical Results of Thermo-Irradiation on the Locally Advanced Hepatoma with or without Hepatic Arterial Chemo-Embolization)

  • 장홍석;윤세철;강기문;유미령;김성환;백남종;윤승규;김부성;신경섭
    • Radiation Oncology Journal
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    • 제12권1호
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    • pp.81-90
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    • 1994
  • Purpose : The aim of this study is to analyze the clinical results of thermo-irradiation treatment for surgically unresectable advanced hepatoma with or without hepatic arterial chemo-embolization (HACE), chemotherapy (CT) and interferon (IFN) therapy. Materials and Methods : Between February 1990 and December 1992, 45 Patients with surgically unresectable advanced hepatomas were treated by thermo-irradiation with or without hepatic arterial chemo-embolization and other treatment modalities. Among them, We analyzed retrospectively 25 patients who received more than three times of hyperthermias. Mean age was 50 years (range : 18-71 years) and male to female ratio was 20 : 5. In the study, treatment was administered as follows : 3 patients received radiation therapy(RT) and hyperthermia (HT). 3 received RT+HT+CT. 3 received RT+HT+HACE. 1 received RT+HT+CT+HACE. 2 received RT+HT+CT+IFN. 10 received RT+HT+HACE+IFN. 3 received RT+HT+CT+HACE+IFN. Radiation therapy was done by a 6 MV linear accelerator Patients were treated with daily fractions of 180 cGy to doses of 11Gy-50Gy (median 30Gy). Local hyperthermia was done by HEH-500C(Omron Co. Japan), 30-45 min/session, 2 sessions/wk and the number of HT sessions ranged from 3 to 17 (median 7 times). 15 patients of 25 were followed by abdominal CT scan or abdominal ultra-sonogram. The following factors were analyzed :Age, histologic grade, sex. number of hyperthermia, total RT dose, hepatic arterial chemo-embolization. Results : Of 25 patients. there were observed tumor regression (partial response and minimal response) in 6 (24$ \% $), no response in 8 (32$ \% $), progression in 1 (4$ \% $) and not evaluable ones in 10 (40$ \% $) radiographically. The over all 1-year survival was 25$ \% $, with a mean survival of 33 weeks. The treatment modes of partial and minimal responsive patients (PR+MR)were as follows : Two were treated with RT+HT+HACE, 2 were done with RT+HT+HACE+IFN Remaining 2 were treated with RT+HT+CT+HACE+IFN. The significant factor affecting the survival rate were RT dose (more than 25 Gy), HACE, number of HT (above 6 times), responsiveness after treatment (PR + MR). Age, sex, histologic differentiation, chemotherapy, interferon therapy were not statistically significant factors affecting the survival rate. Conclusion : Although follow-up duration was short, the thermo-irr3diBtion with/without hepatic arterial chemo-embolization was well tolerated and there were no serious complicatons. In future, it is considered the longer follow up and prospective, well controlled trials should be followed to evaluate the efficacies of survival advantage.

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폐암(肺癌)의 동서의결합치료(東西醫結合治療)에 관(關)한 문헌적(文獻的) 고찰(考察) (Bibliographic Study on the Therapy of Lung Cancer by Integrated Oriental and Western Medicine)

  • 황충연
    • 대한한의학회지
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    • 제16권2호
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    • pp.177-194
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    • 1995
  • 폐암(肺癌)의 치료법(治療法)에 대해 서의치료(西醫治療) 동의치료(東醫治療) 동서의(東西醫) 결합치료(結合治療)에 대해 최근문헌(最近文獻)을 중심(中心)으로 고찰(考察)해 본 결과(結果) 다음과 같은 결론(結論)을 얻었다. 1. 폐암(肺癌)의 서의치료법(西醫治療法)은 소세포암(小細胞癌)(SCLS)과 비소세포암(非小細胞癌)(NSCLS)으로 나누어 분기(分期) 및 증상(症狀)에 따라 수술치료(手術治療) 방사선치료(放射線治療) 화학치료(化學治療)를 단독(單獨)으로 또는 두가지 이상 복합(複合)해서 활용(活用)한다. 2. 폐암(肺癌)의 동의치료(東醫治療)는 초기(初期), 중기(中氣), 말기(末期)로 나누어 부정거사(扶正祛邪), 공보겸시(功補兼施), 기혈쌍보(氣血雙補)의 치법(治法)을 쓰거나 증상(症狀)에 따라 폐비기허형(肺脾氣虛型), 폐열음허형(肺熱陰虛型), 습담어조형(濕痰瘀阻型), 기혈어체형(氣血瘀滯型), 기음양허형(氣陰兩虛型)으로 분(分)하여 변증시치(辨證施治)하거나 혹은 단미(單味) 또는 복방(復方)으로 대증치료(對證治療)를 한다. 3. 폐암(肺癌)에 대(對)한 동서의결합치료(東西醫結合治療)를 함으로써 생존율(生存率)이나 생존(生存)의 질(質)에 있어서 현저(顯著)한 향상(向上)이 있는데 수술후(手術後) 동서결합치료(東西結合治療)는 수술후(手術後) 회복력(回復力)을 촉진(促進)시키고 생존율(生存率)을 높였다. 방사선치료(放射線治療)와의 결합치료(結合治療)는 방사선치료(放射線治療)에 의한 독부작용(毒副作用)을 감소(減少)시켜 치료효과(治療效果)를 높이고 생존율(生存率)을 높였다. 화학요법(化學療法)과 동의결합치료(東醫結合治療)는 화학요법(化學療法)의 독부작용(毒副作用)을 경감(輕減)시키고 생존(生存)의 질(質)과 생존율(生存率)을 높였다. 이상(以上)의 결과(結果)로 보아 폐암(肺癌)의 치료(治療)는 진단(診斷)에서부터 치료(治療)의 전과정(全過程)을 통(通)해서 서의치료(西醫治療)와 동의치료(東醫治療)를 결합(結合)하여 종합치료(綜合治療)를 하므로써 폐암(肺癌)의 치료효과(治療效果)를 높일 수 있는 새로운 치료법(治療法)으로 계속적인 연구(硏究)가 필요(必要)할 것으로 사료(思料)된다.

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한의학의 항종양 면역치료에 관한 연구 -1990년 이후 발표된 실험논문을 중심으로- (Compilation of 104 Experimental Theses on the Antitumor and Immuno-activating therapies of Oriental Medicine)

  • 강연이;김태임;박종오;김성훈;박종대;김동희
    • 동의생리병리학회지
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    • 제17권1호
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    • pp.1-24
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    • 2003
  • This study was done to compile 104 experimental theses which are related to the antitumor and immuno-activating therapies between February 1990 through February 2002. Master's and doctoral theses were dassified by schools, degrees, materials, effects, experimental methods of antitumor and immunoactivity, and results. The following results were obtained from this study : 1. Classifying the theses by the school, 34.6% were presented by Daejeon University, 29.8% by Kyung-hee University and 11.5% by Won-kwang University. Of all theses, 51.0% were aimed for the doctoral degree and 43.3% were for the master's degree. All of three universities have their own cancer centers. 2. Classifying the theses by herb materials, complex prescription accounted for 60.3%, single herb accounted for 24.8% and herbal acupuncture accounted for 14.2%. Considering the key principles of the traditional medicine, complex prescription was much more thoroughly studied than single herb prescription. The results showed that the complex prescription had both antitumor activity and immuno-activating activity, which might reflects on multi-activation mechanisms by complex components. 3. Classifying the theses by the efficacy of herbs examined, in single herb, invigorating spleen and supplementing was 35.5%, expelling toxin and cooling was 29.0%, activating blood flow and removing blood stasis was 12.9%. In herbal acupuncture, invigorating spleen and supplementing was 52.9%, expelling toxin and cooling was 29.4%. In complex prescription, pathogen-free status was 41.9%, strengthening healthy qi to eliminate pathogen was 35.5%, strengthening healthy qi was 22.6%. It is presumed that the antitumor and immunoactivating therapy based on syndrome differentiation is the best way to develop oriental oncology. 4. Classifying the theses by antitumor experiments, cytotoxic effect was 48.1 %, survival time was 48.1 % and change of tumor size was 42.3%. Survival rate was not necessarily correlated with cytotoxicity. These data reflect the characteristic, wholistic nature of the oriental medicine which is based on BRM (biological response modifier). 5. Classifying the theses by immunoactivating experiments, hemolysin titer was 51.0%, hemagglutinin titer was 46.2% and NK cell's activity was 44.2%. In the future studies, an effort to elucidate specific molecular and cellular mechanisms of cytokine production in the body would be crucial. 6. Classifying the theses according to the data in terms of antitumor activity, 50% was evaluated good, 24.0% was excellent, and 15.5% have no effect. In an evaluation of immuno-activating activity, 35.9% was excellent and 18.0% showed a little effect. The index point, as described here, may helps to use experimental data for clinical trials. Changes in index points by varying dosage implicate the importance of oriental medical theory for prescription. 7. In 167 materials, IIP (immunoactivating index point, mean : 3.12±0.07) was significantly higher than AIP(antitumor index point, mean : 2.83±0.07). These data demonstrate that the effect of herb medicine on tumor activity depends more on immunoactivating activity than antitumor activity. This further implies that the development of herbal antitumor drugs must be preceded by the mechanistic understanding of immunoactivating effect. 8. After medline-searching tumor and herb-related articles from NCBI web site, we conclude that most of the studies are primarily focused on biomolecular mechanisms and/or pathways. Henceforth, we need to define the biomolecular mechanisms and/or pathways affected by herbs or complicated prescriptions. 9. Therefore, the most important point of oriental medical oncology is to conned between experimental results and clinical trials. For the public application of herbal therapy to cancer, it is critical to present the data to mass media. 10. To develop the relationship of experimental results and clinical trials, university's cancer clinic must have a long-range plan related to the university laboratories and, at the same time, a regular consortium for this relationship is imperative. 11. After all these efforts, a new type herbal medicine for cancer therapy which is to take care of the long-term administering and safety problem must be developed. Then, it would be expected that anti-tumor herbal acupuncture can improve clinical symptoms and quality of life (QOL) for cancer patients. 12. Finally, oriental medical cancer center must be constructed in NCC (National Cancer Center) or government agency for the development of oriental medical oncology which has international competitive power.

국소진행 두경부 상피세포암에서 선행유도약물요법 후 보조약물요법의 효과 (The Effect of Adjuvant Chemotherapy in Locally Advanced Head and Neck Squamous Cell Carcinoma after Induction Chemotherapy)

  • 김철우;노재경;안중배;박준오;정소영;이석;이혜란;이경희;정현철;김주항;김병수;서창옥;김귀언
    • 대한두경부종양학회지
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    • 제10권1호
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    • pp.13-24
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    • 1994
  • Despite optimal local therapy such as surgery and/or radiotherapy, the long term outcome is poor for patients with advanced squamous cell carcinomma of head and neck, due to frequent loco-regional recurrence and distant metastases. We studied to determine whether the combination chemotherapy, especially as an adjuvant chemotherapy, would improve the survival of these patients. Between January, 1986 and December, 1992, 57 patients with previously untreated, locally advanced squamous cell arcinoma of head and neck were assigned to receive 2-3 cycles of induction chemotherapy consisting of 5-fluorouracil(F) and cisplatin(P) every 3 weeks and standard local therapy such as surgery and/or radiotherapy followed by adjuvant chemotherapy with the same FP regimens. Of the 57 enroled patients, 45 patients were evaluable. The obtained results were as following: 1) Among 45 evaluable patients, 18 patients finished all treatment protocol including adjuvant chemotherapy and 27 patients had no adjuvant chemotherapy. The difference of age, sex, performance status, disease stage, and tumor differentiation was not significant statistically between adjuvant chemotherapy group and no-adjuvant chemotherapy group. 2) After induction chemotherapy, 7/45(15.4%), 30/45(67%) achieved complete remission and partial remission respectively with 82.4% overall response rates in entire patients. 3) The 4year progression free survival was 43.3% in adjuvant chemotherapy group and 24.1% in no-adjuvant chemotherapy group(p>0.05). The 4year overall survival was 56.9% and 25.5% respectively(p>0.05). There was no significant different in the patterns of local recurrence and distant metastasis between the two groups. 4) Adverse reactions from combination chemotherapy included nausea, vomiting, mucositis, diarrhea and hematologic bone marrow depression. These were mild and tolerated by patients, and these was no episode of any life threatening toxicities. In conclusion, adjuvant chemotherapy after induction chemotherapy and local therapy did not show statistically significant survival improvement, but there was trend of prolongation of survival when compared to no adjuvant chemotherapy. Thus, large scale phase III randomized controlled studies are strongly recommended.

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${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명 (Establishment and Characterization of Bone Marrow Mesenchymal Stromal/Stem Cells (MSCs) Derived from ${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) Pig)

  • 옥선아;오건봉;황성수;임석기;김영임;박진기
    • 한국수정란이식학회지
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    • 제28권3호
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    • pp.281-287
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    • 2013
  • A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

악성 타액선 종양의 방사선 치료 성적에 대하여 -원자력 병원의 10년 경험 ($1975.1{\sim}1984.12$)- (Primary Radiation Therapy of Malignant Salivary Gland Tumors by Conventional Megavoltage Irradiation -Korea Cancer Center Hospital-)

  • 조철구;고경환;류성렬;박영환;박우윤;심윤상;오경균
    • Radiation Oncology Journal
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    • 제8권1호
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    • pp.35-43
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    • 1990
  • 1975년 1월부터 1984년 12월까지 원자력병원 치료 방사선과에서 악성 타액선 종양으로 방사선 치료를 받은 58명의 환자를 대상으로 하여 이들의 생존율을 후향적으로 분석하였다. 이들은 수술후 재발했거나, 수술이 불가능한 환자들이었다. 58명의 환자중 mucoepidermoid carcinoma를 가진 환자가 $43.1\%$, adenoid cystic carcinoma를 가진 환자는 $41.3\%$였다. 주 타액선 종양의 5년 보험생존율은 $68.2\%$, 10년 생존율은 $31.8\%$였으나, 무병생존율은 각각 $43.2\%\;13.0\%$로써 치료 후 재발된 상태에서도 비교적 오래 산다는 것을 알 수 있었다. TNM staging에 의한 생존율도 $T_1$의 5년 생존율이 $86.5\%,\;T_2+T_3$$40.0\%,\;T_4$$0\%$로, T stage가 높아지면 질수록 생존율도 현저히 감소하였다. 병리조직학적 관점에서 볼 때, adenoid cystic carcinoma의 5년 무병생존율은 $40.1\%$로써, mucoepidermoid ca.의 $49.8\%$보다 낮았으나, 전체적인 생존율은 $77.3\%$로써, mucoepidermoid ca.의 $51.5\%$보다 현저히 높았다. 따라서, adenoid cystic carcinoma는 치료실패후 병을 가진 상태에서도 상당 기간 생존할 수 있다는 것을 알았으며, 평균 생존기간은 2년 이었다. 또한 mucoepidermoid ca.인 경우에는 세포의 분화정도에 따라 생존율이 달라졌는데, 저등도 분화세포의 5년 생존율이 $78.8\%$로 고등도 분화세포의 $38.2\%$보다 거의 2배나 높았다. 암의 위치와 성별에 따른 생존율의 차이는 없었다. Minor salivary gland tumor는 6명으로 5년 보험생존률은 $32.3\%$였다. 따라서 주 타액선 종양의 생존율에 영향을 끼칠 수 있는 예후 인자는 1) 병리조직학적 세포종류, 2) T와 N stages (AJCC), 3) mucoepidermoid carcinoma에 있어서 분화 정도 였다.

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초기 계대 인간 배아줄기세포의 해동 후 효율적인 배양 방법 (Efficient Culture Method for Early Passage hESCs after Thawing)

  • 백진아;김희선;설혜원;서진;정주원;윤보애;박용빈;오선경;구승엽;김석현;최영민;문신용
    • Clinical and Experimental Reproductive Medicine
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    • 제36권4호
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    • pp.311-319
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    • 2009
  • 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.

Hypoxia-dependent mitochondrial fission regulates endothelial progenitor cell migration, invasion, and tube formation

  • Kim, Da Yeon;Jung, Seok Yun;Kim, Yeon Ju;Kang, Songhwa;Park, Ji Hye;Ji, Seung Taek;Jang, Woong Bi;Lamichane, Shreekrishna;Lamichane, Babita Dahal;Chae, Young Chan;Lee, Dongjun;Chung, Joo Seop;Kwon, Sang-Mo
    • The Korean Journal of Physiology and Pharmacology
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    • 제22권2호
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    • pp.203-213
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    • 2018
  • Tumor undergo uncontrolled, excessive proliferation leads to hypoxic microenvironment. To fulfill their demand for nutrient, and oxygen, tumor angiogenesis is required. Endothelial progenitor cells (EPCs) have been known to the main source of angiogenesis because of their potential to differentiation into endothelial cells. Therefore, understanding the mechanism of EPC-mediated angiogenesis in hypoxia is critical for development of cancer therapy. Recently, mitochondrial dynamics has emerged as a critical mechanism for cellular function and differentiation under hypoxic conditions. However, the role of mitochondrial dynamics in hypoxia-induced angiogenesis remains to be elucidated. In this study, we demonstrated that hypoxia-induced mitochondrial fission accelerates EPCs bioactivities. We first investigated the effect of hypoxia on EPC-mediated angiogenesis. Cell migration, invasion, and tube formation was significantly increased under hypoxic conditions; expression of EPC surface markers was unchanged. And mitochondrial fission was induced by hypoxia time-dependent manner. We found that hypoxia-induced mitochondrial fission was triggered by dynamin-related protein Drp1, specifically, phosphorylated DRP1 at Ser637, a suppression marker for mitochondrial fission, was impaired in hypoxia time-dependent manner. To confirm the role of DRP1 in EPC-mediated angiogenesis, we analyzed cell bioactivities using Mdivi-1, a selective DRP1 inhibitor, and DRP1 siRNA. DRP1 silencing or Mdivi-1 treatment dramatically reduced cell migration, invasion, and tube formation in EPCs, but the expression of EPC surface markers was unchanged. In conclusion, we uncovered a novel role of mitochondrial fission in hypoxia-induced angiogenesis. Therefore, we suggest that specific modulation of DRP1-mediated mitochondrial dynamics may be a potential therapeutic strategy in EPC-mediated tumor angiogenesis.