• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1349-1353
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• 2012

Increasing evidence has revealed that thy-1 was a potential stem cell marker of liver cancer, but no data have been shown on how thy-1 regulates the pathophysiology of liver cancer, such as proliferation, apoptosis, invasion and migration. We previously demonstrated that thy-1 was expressed in about 1% of hepg2 cells, thy-1+hepg2 cells, but not thy-1-, demonstrating high tumorigenesis on inoculation $0.5{\times}10^5$ cells per BACA/LA mouse after 2 months. In the present study, our results showed that higher expression of thy-1 occurs in 72% (36/50 cases) of neoplastic hepatic tissues as compared to 40% (20/50 cases) of control tissues, and the expression of thy-1 is higher in poorly differentiated liver tumors than in the well-differentiated ones. In addition, thy-1 expression was detected in 85% of blood samples from liver cancer patients, but none in normal subjects or patients with cirrhosis or hepatitis. There was a significant negative correlation between thy-1expression and E-cadherin expression (a marker of invasion and migraton), but not between thy-1 expression and AFP expression in all the liver cancer and blood samples. We further investigated the relationship between thy-1 and E-cadherin in liver cancer hepg2 cell line which was transfected with pReceiver-M29/thy-1 eukaryotic expression vector followed by aspirin treatment. Lower expression of E-cadherin but higher expressions of thy-1 were detected in hepg2 cells transfected with pReceiver-M29/thy-1. Taken together, our study suggested that thy-1 probably regulates liver cancer invasion and migration.

• Asian-Australasian Journal of Animal Sciences
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• 제26권7호
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• pp.935-944
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• 2013

Essential oils have been shown to favorably effect in vitro ruminal fermentation, but there are few in vivo studies that have examined animal responses. The objective of this study was to evaluate the effects of thyme (THY) and cinnamon (CIN) essential oils on feed intake, growth performance, ruminal fermentation and blood metabolites in feedlot calves fed high-concentrate diets. Twelve growing Holstein calves ($213{\pm}17kg$ initial BW) were used in a completely randomized design and received their respective dietary treatments for 45 d. Treatments were: 1-control (no additive), 2-THY (5 g/d/calf) and 3-CIN (5 g/d/calf). Calves were fed ad libitum diets consisting of 15% forage and 85% concentrate, and adapted to the finishing diet by gradually increasing the concentrate ratio with feeding a series of transition diets 5 wk before the experiment started. Supplementation of THY or CIN did not affect DMI and ADG, and feed efficiency was similar between treatment groups. There were no effects of additives on ruminal pH and rumen concentrations of ammonia nitrogen and total VFA; whereas molar proportion of acetate and ratio of acetate to propionate decreased, and the molar proportion of propionate increased with THY and CIN supplementation. Rumen molar concentration of butyrate was significantly increased by adding CIN compared to control; but no change was observed with THY compared with control group. No effects of THY, or CIN were observed on valerate, isobutyrate or isovalerate proportions. Plasma concentrations of glucose, cholesterol, triglyceride, urea-N, ${\beta}$-hydroxybutyrate, alanine aminotransferase and aspartate aminotransferase were not changed by feeding THY or CIN. Results from this study suggest that supplementing a feedlot finishing diet with THY or CIN essential oil might be useful as ruminal fermentation modifiers in beef production systems, but has minor impacts on blood metabolites.

• Archives of Pharmacal Research
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• 제21권4호
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• pp.398-405
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• 1998

We investigated the effects of triterpene acids (TAs), ursolic acid (UA) and oleanolic acid (OA), on the induction of proliferation and differentiation of normal rat mammary epithelial cells (RMEC) or organoids cultured in Matrigel or primary culture system. To elucidate the effects, we tested their differentiation inducing activities with intercellular communication ability, cell cycle patterns, induction of apoptosis, and morphological differentiation in the three dimensional extracellular culture system. To study the changes of RMEC subpopulation in culture, the cultured cells were isolated, immunostained with peanut lectin (PNA) and anti-Thy-1.1 antibody and then analyzed with flow cytometry. Four different subpopulations, such as PNA and Thy-1.1 negative cells (B-), PNA positive cells (PNA+), Thy-1.1 positive cells (Thy-1.1+), PNA and Thy-1.1 positive cells (B+), were obtained and the size of each subpopulation was changed in culture with time in the presence of TAs. Intercellular communication was observed in culture for 7 days in TAs-treated cells, but not in culture for 4 days with scrape-loading dye transfer technique. $G_2$/M phase cells and the number of apoptotic population were increased in TAs-treated groups in cell cycle analyses. S phase fractions were reduced and the change of $G_1$ phase cells was not observed. The colonies with distinct multicelfular structures, such as stellate, ductal, webbed, squamous, lobulo-ductal colonies, were observed in Matrigel culture and the frequencies of each colony were changed in the presence of TAs. These results suggest that UA and OA have differentiation inducing effects on rat mammary epithelial cells in primary or in Matrigel culture.

• 생명과학회지
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• 제10권1호
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• pp.37-44
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• 2000

The mammary gland contains a subpopulation of epithelial cells with large proliferative potentials which are the likely targets for carcinogens. These clonogenic cells can proliferate and differentiate into functional glandular structures. Rat mammary epithelial cells (RMEC) were isolated and characterized in vitro. By flow cytometry of RMEC stained with fluorescein isothiocyanate-peanut agglutinin(PNA) and phycoerythrin anti-Thy-1.1 monoclonal antibody, it was possible to four cell subpopulations from 7-8 week old F344 female rat mammary glands: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). When single PNA+ cells were isolated and cultured in Matrigel with irradiated (∼50 Gray) 3T3 fibroblast feeder layer, they gave rise to multicellular clonal structures of three types: alveolar, foamy alveolar, and squamous colonies. The developed structures were similar to the mammary glands in vivo. These results suggest that some of PNA+ cells possesses many of the characteristics of multipotent clonogenic stem-like cells.

• 농업과학연구
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• 제19권1호
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• pp.51-64
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• 1992

본(本) 실험(實驗)은 수유중(泌乳中)인 흰쥐를 대상(對象)으로 갑상선기능(甲狀腺機能)이 체중(體重)과 수유(泌乳)에 미치는 영향(影響)을 구명(究明)코져 실시(實施)하였는 바 체중 250g 정도(程度)의 정상분만(正常分娩)한 암흰쥐 105마리를 thyroxine 처리군(處理群)(THY.군(群)), propylthiouracil 처리군(處理群)(PTU군(群)) 및 정상대조(正常對照) 군(群)(CON.군(群))의 3개군(個群)으로 나누어 35마리씩 배치(配置)한 다음 THY.군(群)에서는 L-thyroxine을 체중 100g당(當) $30{\mu}m$씩 3일(日) 간격(間隔)으로 주사(注射)하였고 PTU군(群)에서는 음수(飮水)를 0.03%의 propylthiouracil 용액(溶液)으로 만들어 자유급수(J自由給水)시켰다. 또한 각(各) 군별(群別)로 시간(時間)의 경과(經過)에 따라 3, 6, 9, 12, 15, 18 및 21일(日)에 체중(體重)을 측정(測定)하였고 도살(屠殺) 후(後)에 갑상선(甲狀腺)의 중량측정(重量測定), 갑상선(甲狀腺)과 유선(乳腺)의 조직학(組織學)적 검색(檢索)을 실시(實施)하였으며 아울러 혈청중(血淸中) prolactin의 농도(濃度)를 측정(測定)하여 비교(比較) 검토(檢討)한 결과(結果)는 다음과 같다. 1. 체중(體重)은 처리후(處理後) 3일(日)과 6일(日) 그리고 15일(日)과 18일(日)에서 비교군간(比較群間) 유의성(有意性)이 인정(認定)되는 변화(變化)를 보였는데 PTU군(群)이 CON.군(群)과 THY.군(群)에 비(比)하여 낮은 중량치(重量値)를 나타냈다. 2 갑상선(甲狀腺)의 중량(重量)은 처리(處理) 9일(日) 후(後)부터 비교군간(比較群間)에 유의성(有意性)이 인정(認定)되었는데 PTU군(群)은 CON.군(群)과 THY.군(群)에 비(比)하여 고도(高度)의 유의성(有意性)이 인정(認定)되는 높은 중량치(重量値)를 나타냈다. 3. 갑상선(甲狀腺)의 조직상(組織象)은 PTU군(群)의 경우 처리(處理) 6일(日) 후(後)부터 농포상피세포(濃胞上皮細胞)가 증식(增殖).비대(肥大)를 동반(同伴)한 원주화(圓柱化) 현상(現象)이 나타나기 시작하였고 이러한 현상(現象)은 수유말기(泌乳末期)까지 계속되었다 THY.군(群)은 처리(處理) 9일(日) 후(後)부터 핵농축(核濃縮)을 동반(同伴)한 농포상피세포(農胞上皮細胞)가 편평화(扁平化)되는 퇴행성(退行性) 변화(變化)를 나타냈다. 4. CON.군(群)의 prolactin 농도(濃度)는 처리(處理) 18일후(日後)에 최고치(最高値)를 보였고 THY.군(群)은 처리(處理) 12일후(日後)에 고도(高度)의 유의성(有意性)이 인정(認定)되는 높은 농도(濃度)를 나타냈으며 PTU군(群)은 전(全) 실험기간(實驗其間)동안 다른 실험군(實驗群)에 비(比)하여 낮은 농도(濃度)를 나타냈다. 5. 유선(乳腺)의 조직학적(組織學的) 변화(變化)는 CON.군(群)과 THY.군(群)은 처리(處理) 12일후(日後)까지는 유선상피세포(乳腺上皮細)가 입방화(立方化)되는 조직상(組織象)을 나타냈으나 처리(處理) 후(後) 15일(日)부터는 THY.군(群)이 CON.군(群)보다 유선조직(乳腺組織)의 분화(分化)가 빠른 속도(速度)로 진행(進行)되었으며 PTU군(群)은 처리(處理) 후(後) 시간(時間)이 경과(經過)할수록 유선조직(乳腺組織)의 퇴행성(退行性) 변화(變化)가 야기(惹起)되어 처리(處理) 15일후(日後)부터는 유선상피세포(乳腺上皮細胞)의 탈락현상(脫落現象)과 유선포(乳腺胞)의 위축(萎縮)이 인지(認知)되는 조직소견(組織所見)이었다. 6. 신생자(新生仔)의 체중(體重)은 처리(處理) 후(後) 시간(時間)이 경과(經過)할수록 모든 실험군(實驗群)이 점차(漸次) 증가(增加)하였으나 PTU군(群)은 다른 실험군(實驗群)에 비(比)하여 유의성(有意性)이 인정(認定)되는 낮은 체중치를 나타냈다.

• 한국식품영양과학회지
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• 제24권3호
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• pp.470-486
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• 1995

유선에 존재하는 유선 상피 모세포(mammary epithelial stem cells)의 존재 증거, 정상 조직에서 이들의역할, flow cytometry 및 면역 염색법에 의한 세포 분리, 세포 기질 단백질을 이용한 삼차원적 세포 배양에서의 증식 등을 요약한다. 유선의 실질 조직에 상피 모세포가 존재한다는 것은 여러 형태의 이식 실험에서 설명되었고 또 모세포의 표현형적 특징들은 여러 가지의 monoclonal antibodies에 의해 논증되었다. 이들 연구의 결과들은 유선의 모세포군이 end bud와 유선의 기저층(basal layer)에 존재한다고 제시하고 있다. 이들을 분리, 동정하기 위해 FITC-PNA와 PE-Thy-1.1 항체와 같은 세포 표지자를 이용하여 유선 상피 세포를 4군으로 나눌 수 있었다. FITC-PNA에만 양성 반응을 보인 PNA+ 세포군, PE-Thy-1.1에만 양성 반응을 보인 Thy-1.1+ 세포군, 이들 두 표지자에 양성 반응을 보인 B+ 세포군, 그리고 양쪽에 음성 반응을 보인 B- 세포군이었는데 이들을 flow cytometry로 분리하고 생체에 이식 실험을 하였을 때 PNA+ 세포군이 유선 모세포들을 가장 많이 가진 것으로 확인되었다. 그리고 유선 상피세포로 이루어진 유선 조직 절편(organoids) 이들 상피세포군을 세포외기질 단백질체인 Matrigel 내에서 배양한 결과 a) stellate, b) duct, c) web, d)squamous, e) lobuloduct 등 5종류의 다세포 구조물이 생성됨을 확인하였다. 이들 중 편평상피화생의 구조물은 정상적인 유선 조직에서는 나타나지 않는 구조물인데 all-trans retinoic acid를 처리하였을 때 배지의 조정에 따라 다소 차이는 있으나 대부분 이들 편평상피화생의 생성이 억제됨을 확인하였다. 이상의 결과로 보아 본 연구에 이용된 생체 이식법 및 삼차원적 세포외기질 세포 배양법이 상피세포의 성장, 분화 및 모세포 연구에 유용하게 이용될 수 있으리라 사료된다.

• 한국콘텐츠학회:학술대회논문집
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• 한국콘텐츠학회 2018년도 춘계 종합학술대회 논문집
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• pp.551-552
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• 2018

K/BxN serum can induce arthritis in normal mice because of abundant autoantibodies that trigger an innate inflammatory response in joints. To determine whether IL-17 is involved in the pathogenesis of serum-induced arthritis, we injected wild-type and $IL-17^{-/-}$ mice with K/BxN serum and evaluated them for signs of arthritis. Unlike wild-type mice, $IL-17^{-/-}$ mice did not show any signs of arthritis. IL-17 was produced predominantly by $CD3^-CD4^-gdTCR^-NK1.1^-Sca1^{int}Thy1^{hi}$ cells residing in the inflamed synovial tissue. When synovial cells extracted from normal joints were stimulated with IL-23 or autoantibody-containing immune complexes, a substantial fraction of $Sca1^{int}Thy1^{hi}$ cells produced IL-17. Thus, we have identified a novel population of IL-17-producing innate synovial cells that play a crucial role in the development of K/BxN serum-induced arthritis.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 International Symposium of Silkworm/Insect Biotechnology and Annual Meeting of Korea Society of Sericultural Science
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• pp.145-146
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• 2003

Overproduction of transforming growth factor (TGF)-$\beta$l has been implicated in the pathogenesis of fibrotic diseases. TGF-$\beta$l plays a crucial role in the accumulation of extracellular matrix (ECM) in human and experimental glomerular diseases. However, it remains unclear whether inhibition of TGF- $\beta$l overproduction would suppress TGF- $\beta$l induced ECM accumulation. To inhibit the overproduction of TGF- $\beta$l in experimental glomerulonephritis induced by anti-Thy 1.1 antibody, we introduced antisense oligodeoxynucleotides (ODN) fur TGF- $\beta$l into the nephritic kidney by the HVJ-liposome-mediated gene transfer method. (omitted)

• 한국수정란이식학회지
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• 제33권4호
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• pp.327-336
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• 2018

To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.

• 한국발생생물학회지:발생과생식
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• 제15권1호
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• pp.41-52
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• 2011

In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/$m{\ell}$ BMP4 or 0.01 ${\mu}M$ RA. In the BMP4-treated group, GFR-${\alpha}1$ and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-${\alpha}1$ and CD90/Thy-1 expression in the EBs of mSSCs Induced pluripotent stem cells, Mouse embryonic stem cells, Multipotent spermatogonial stem cells, Germ cell lineage, Differentiation potential. was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1 and immunocytochemistry using GFR-${\alpha}1$ antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.