• 한국정보통신학회:학술대회논문집
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• 한국정보통신학회 2014년도 춘계학술대회
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• pp.822-825
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• 2014

시험내 수초화 시스템을 만들기 위해 쥐에서 슈반세포와 뉴런세포의 공동 배양이 완성되었다. 슈반세포와 뉴런 세포는 각각 쥐의 배아(임신 15일)의 척수신경절로 부터 분리되었다. 이 방법은 4단계로 이루어져 있다. 1단계는 쥐배아의 척수신경절를 부유시키는 단계, 2단계는 유사분열억제제를 첨가하는 단계, 3단계는 척수신경절 세포를 순수 분리하는 단계, 4단계는 척수신경절 세포에 슈반세포를 첨가하는 단계이다. 우리는 단기간 내에 고 순도의 수초화 군을 생성하였으며 이렇게 생성된 수초화 단백질을 수초 기본 단백질(myelination basic protein)의 항체를 이용하여 확인하였다.

• 농업과학연구
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• 제23권2호
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• pp.212-218
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• 1996

본 연구는 체외에서 성숙 및 수정된 돼지 체외 수정란을 두 종류의 서로 다른 배양액에 단층난구 세포와 공배양하였을 경우 배발달에 미치는 영향을 조사하기 위하여 실시하였다 본 연구에서 얻은 결과를 요약하면 다음과 같다. 1. 단층난구세포와 함께 초기체외수정란을 Ham's F-10에서 배양한 결과 2-, 4-, 8~16-세포기, 상실배/배반포기로 발달하는 비율은 각각 48.7%, 40.5%, 34.8% 및 17.0% 였으며, TCM-HEPES에서는 각각 48.3%, 35.6 %, 22.1% 및 13.4%였다. 2. 단층난구세포와 공배양을 하지 않고 단순하게 배양액으로만 배양하였을 때 상실배/배반포기까지의 발달률은 0.0~1.0%로서 극히 저조하였다. 특히, 대부분의 수정란이 4-세포기 단계에서 발달이 정지되었다. 3. 돼지 체외수정란을 단층난구세포와 공배양하였을 때 단순배양한 결과보다 유의 적으로 높은 상실배/배반포기의 발달률을 나타냈으며 (P<0.05), 두 배양액간에는 유의차가 인정되지 않았다.

• 한국약용작물학회:학술대회논문집
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• 한국약용작물학회 2011년도 추계학술발표회
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• pp.326-327
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• 2011

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2003년도 제3회 발생공학 국제심포지움 및 학술대회
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• pp.77-77
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• 2003

• 미생물학회지
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• 제47권3호
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• pp.179-187
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• 2011

고찰을 통해 난배양성 토양세균의 특징과 배양에 성공한 사례 및 성공하기 위해 알아야 될 지식들이 무엇인지에 대해 기술하였다. 먼저 배지는 목적한 세균이 토양에서 느리게 성장하다가 실험실의 빠른 성장조건으로 전환하도록 알맞게 선택되어야 하는데 일반적으로 기질, 질소 및 인 등의 농도를 낮게 조절해야 한다. 새로운 배지를 만들기 위해서는 분자생태학적 연구도 병행되어야 한다. 세균 세포 간 음성적 상호작용을 줄이기 위해 평판배양 시 접종량도 평판 당 세포수가 50개 이하로 조절해야 한다. pH나 염농도 같은 성장조건은 실제 환경조건과 맞춰야 하며 배양온도는 낮거나 다양하게 그리고 배양기간은 길게 잡아야 한다. 새로운 배지에서 분리될 수많은 토양 미생물 콜로니들 중에서 단지 몇 개만이 난배양성이므로 이들이 기존에 배양이 되지 않았던 미생물인지를 신속 정확히 검출하는 방법이 필요하다. 또한 많은 토양세균들이 군집 내에서 서로 협력하며 살아가기 때문에 공동배양이나 상등액을 이용해서 토양 미생물을 농화증식하고 이를 순수 분리하면 배양에 성공할 수 있을 것이다.

• International Journal of Oral Biology
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• 제33권3호
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• pp.113-116
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• 2008

Cementum is the mineralized tissue of the tooth. It is similar to bone in several aspects but it differs from bone. Human bone marrow stromal cells (BMSC) and human cementum derived cells (HCDC) (10,000 $cells/cm^2$) were plated in 6 well plates as feeder cells. The next day, mouse bone marrow cells (1.5 million $cells/cm^2$) were added. One group of these plates were incubated in serum-free conditioned medium (SFCM) generated from BMSC or HCDC supplemented with 2% FBS, parathyroid hormone (PTH), 1, 25 dihydroxyvitamin $D_3$ (Vit. $D_3$) and dexamethasone, or plain medium with the same supplements. Another group of plates were cocultured with BMSC or HCDC in plain medium supplemented with 2% FBS, PTH, Vit. $D_3$ and dexamethasone. Plates grown without SFCM or coculture were used as controls. After 10 days, the cells were stained for tartrate-resistant acid phosphatase (TRAP). BMSC were found to support osteoclast formation under normal conditions. This was inhibited however by both SFCM generated from HCDC and also by coculture with HCDC. In addition, HCDC themselves did not support osteoclast formation under any conditions. Our results thus indicate that HCDC do not support osteoclast formation in vitro and that soluble factor (s) from HCDC may inhibit this process. In addition, we show that this inhibition also involves an active mechanism that is independent of osteoprotegerin, a feature that may distinguish cementoblasts from other cells present in periodontium.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3605-3610
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• 2016

Background: Histone acetylation in chromatin structures plays a key role in regulation of gene transcription and is strictly controlled by histone acetyltransferase (HAT) and deacetylase (HDAC) activities. HDAC deregulation has been reported in several cancers. Materials and Methods: The expression of 10 HDACs (including HDAC class I and II) was studied by quantitative reverse transcription-PCR (qRT-PCR) in a cohort of mesenchymal stem cells (MM-MSCs) from 10 multiple myeloma patients with a median age 60y. The results were compared with those obtained for normal donors. Then, a coculture system was performed between MM-MSCs and u266 cell line, in the presence or absence of sodium butyrate (NaBT), to understand the effects of HDAC inhibitors (HDACi) in MM-MSCs on multiple myeloma cases. Also, the interleukin-6 (IL-6) and vascular endothelial growth factor (VEGFA) gene expression level and apoptotic effects were investigated in MM-MSCs patients and control group following NaBT treatment. Results: The results indicated that upregulated (HDACs) and downregulated (IL6 and VEGFA) genes were differentially expressed in the MM-MSCs derived from patients with multiple myeloma and ND-MSCs from normal donors. Comparison of the MM-MSCs and ND-MSCs also showed distinct HDACs expression patterns. For the first time to our knowledge, a significant increase of apoptosis was observed in coculture with MM-MSCs treated with NaBT. Conclusions: The obtained findings elucidate a complex set of actions in MSCs in response to HDAC inhibitors, which may be responsible for anticancer effects. Also, the data support the idea that MSCs are new therapeutic targets as a potential effective strategy for MM.

• Toxicological Research
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• 제8권2호
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• pp.191-203
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• 1992

The immunosuppressive effects of safrole were studied in female BALB/c mouse. Mice were given 100,200and 400mg safrole/kg daily for 14days and evaluated on day 15. The day 4 immunogloblin-M antibody response to T-dependent antigen, sheep red blood cells (SRBC) was inhibited dose-dependently in all doses studied. In vitro antibody response to polyclonal antigen, lipopolysaccharide (LPS) by spleen cell suspensions from safrole-treated mice were also significantly inhibited. When safrole was treated for 14days to mice, and mitogen-induced proliferation of splenocytes were assayed on day 15, there were significant suppression of responses to B-cell mitogen, LPS and T-cell mitogen concanavalin A(Con A) at a dose of 400mg safrole/kg. Direct addition of safrole on the splenocyte culture also produced a dose dependent suppression on in vitro antibody response to LPS, and mitogen-induced lymphoproliferatin at doses of 100,200,400 and 800${\mu}M$ safrole. The role of metabolic activation in safrole-induced suppression of in vitro antibody response was studied using splenocyte-hepatocyte coculture system. The suppression of in vitro antibody respose to LPS by safrole was not altered when safrole were incubated in the splenocyte-hepatocyte system for 4hr as compared with direct addition of safrole in splenocytes culture. Neither the addition of salicylamide, sulfotransferase inhibitor, nor the addation of inorganic sulfate, sulfation cofactor to the splenocyte-hepatocyte coculture, altered the suppression of antibody response by safrole. These results suggest that the immunosuppression by safrole may not by produced by the reactive metabolites which are mediated in carcinogenesis of safrole.

• Archives of Pharmacal Research
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• 제22권4호
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• pp.340-347
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• 1999

Dendritic cells (DCs) are potent professional antigen-presenting cells (APC) capable of inducing the primary T cell response to antigen. Although tumor cells express target antigens, they are incapable of stimulating a tumor-specific immune response due to a defect in the costimulatory signal that is required for optimal activation of T cells. In this work, we describe a new approach using tumor-DC coculture to improve the antigen presenting capacity of tumor cells which does not require a source of tumor-associated antigen. Immunization of a weakly immunogenic and progressive tumor cocultured with none marrow-derived DCs generated an effective tumor vaccine. Immunization with the cocutured DCs was able to induce complete protectiv immunity against tumor challenges and was effective for the induction of tumor-specific CTL (cytotoxic T lymphocyte) activity. Furthermore, high NK cell activity was observed in mice in which tumors were rejected. In addition, immunization with tumor-pulsed DC s induced delayed tumor growth, but not tumor eradication in tumor-bearing mice. Our results demonstrate that coculture of DCs with tumors generated antitumor immunity due to the NK cell activation as well as tumor-specific T cell. This approach would be used for designing tumor vaccines using DCs when the information about tumor antigens is limited.

• Journal of Veterinary Science
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• 제22권3호
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• pp.25.1-25.16
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• 2021

Background: Malignant lymphoma is the most common hematopoietic malignancy in dogs, and relapse is frequently seen despite aggressive initial treatment. In order for the treatment of these recurrent lymphomas in dogs to be effective, it is important to choose a personalized and sensitive anticancer agent. To provide a reliable tool for drug development and for personalized cancer therapy, it is critical to maintain key characteristics of the original tumor. Objectives: In this study, we established a model of hybrid tumor/stromal spheroids and investigated the association between canine lymphoma cell line (GL-1) and canine lymph node (LN)-derived stromal cells (SCs). Methods: A hybrid spheroid model consisting of GL-1 cells and LN-derived SC was created using ultra low attachment plate. The relationship between SCs and tumor cells (TCs) was investigated using a coculture system. Results: TCs cocultured with SCs were found to have significantly upregulated multidrug resistance genes, such as P-qp, MRP1, and BCRP, compared with TC monocultures. Additionally, it was revealed that coculture with SCs reduced doxorubicin-induced apoptosis and G2/M cell cycle arrest of GL-1 cells. Conclusions: SCs upregulated multidrug resistance genes in TCs and influenced apoptosis and the cell cycle of TCs in the presence of anticancer drugs. This study revealed that understanding the interaction between the tumor microenvironment and TCs is essential in designing experimental approaches to personalized medicine and to predict the effect of drugs.