• Clinical and Experimental Reproductive Medicine
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• 제50권4호
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• pp.244-252
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• 2023

Objective: We evaluated the efficacy of the newly developed optimized in vitro culture (OIVC) dish for cultivating preimplantation mouse embryos. This dish minimizes the need for mineral oil and incorporates microwells, providing a stable culture environment and enabling independent monitoring of individual embryos. Methods: Mouse pronuclear (PN) zygotes and two-cell-stage embryos were collected at 18 and 46 hours after human chorionic gonadotropin injection, respectively. These were cultured for 120 hours using potassium simplex optimized medium (KSOM) to reach the blastocyst stage. The embryos were randomly allocated into three groups, each cultured in one of three dishes: a 60-mm culture dish, a microdrop dish, and an OIVC dish that we developed. Results: The OIVC dish effectively maintained the osmolarity of the KSOM culture medium over a 5-day period using only 2 mL of mineral oil. This contrasts with the significant osmolarity increase observed in the 60-mm culture dish. Additionally, the OIVC dish exhibited higher blastulation rates from two-cell embryos (100%) relative to the other dish types. Moreover, blastocysts derived from both PN zygotes and two-cell embryos in the OIVC dish group demonstrated significantly elevated mean cell numbers. Conclusion: Use of the OIVC dish markedly increased the number of cells in blastocysts derived from the in vitro culture of preimplantation mouse embryos. The capacity of this dish to maintain medium osmolarity with minimal mineral oil usage represents a breakthrough that may advance embryo culture techniques for various mammals, including human in vitro fertilization and embryo transfer programs.

• 환경생물
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• 제26권1호
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• pp.30-35
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• 2008

곰피추출물의 CCD-986sk cell line monolayer (human fibroblast, KCBL-21947)에 대한 피부세포 생리활성효과를 측정하고, 또한 곰피추출물의 Clone M-3 mouse melano-cyte cell line에 대한 melanin formation 저해효과를 측정하기 위해 in vitro레벨에서 실험을 실시하였다. 곰피는 다년생 갈조류의 일종으로 이 종은 한국 연안해역에서 중요한 1차생산자의 역할을 담당하고 있는 종이며 연안 생태계에 있어서 생태학적으로나 수산업상 매우 중요한 위치를 점유하고 있다. 최근 이러한 대형 갈조류에 관한 연구는 주로 생태학적인 관점에서 이루어져 왔으며(Notoya and Aruga 1990), 유주자의 발아와 핵분열(Yabu and Notoya 1985) 등에 관한 연구가 주로 일본의 연구자들에 의하여 보고된 바 있고, 국내에서는 Park et al. (1994)이 부산만 인근해역 곰피의 생장과 연령 조성에 관한 연구를 수행하였고, 순간접착제를 이용한 곰피의 이식 등에 관한 연구(최 등 2002)가 보고된 바 있다. 그러나 지금까지 곰피의 식품 또는 약품제제로서의 기능성을 밝히는 연구는 아직 미미한 실정으로 이에 대한 체계적인 연구가 부족한 실정이다. 따라서 본 연구는 곰피추출물에 풍부하게 함유된 fucoidan, L-Ascorbic 산 및 collagen 등에 주목하여 곰피의 피부노화방지제로서의 가능성을 체계적으로 연구하기 위한 일환으로 우선 곰피추출물이 피부재생, 주름살제거 및 피부미백작용에 효과를 나타내는지 확인하기 위해 CCD-986sk cell line 및 Clone M-3 mouse melanocyte cell line을 이용하여 in vitro 수준에서 피부세포 생리활성 효과와 melanin formation 저해효과를 측정하였다. 그 결과 CCD-986sk cell line에 대한 세포생리활성 효과 실험에서 시험군인 곰피추출물 투여군은 대조군에 비해 강한 세포생리활성 효과를 나타내어 유의성 있는 실험결과를 보여주었고, Clone M-3 mouse melanocyte cell에 대한 melanin pigment 저해활성 효과 실험에서도 곰피추출물은 대조군에 비해 melanin pigment 생합성을 유의성 있게 감소시키는 실험결과를 나타내어 매우 흥미로운 결과를 보여주었다. 여러 가지 보고에 의하면 그동안 미국, 영국, 일본 등 구미 선진국의 과학자들은 intrinsic problems (피부구조 및 생리기능의 노쇠)과 extrinsic problems (자외선노출 및 스트레스)을 skin aging의 근본원인으로 규정짓고 피부 콜라겐과 엘라스틴을 재생시켜 피부 horny layer의 주름을 제거시키는 수 많은 종류의 chemical drugs를 개발하였고 현재도 개발 중에 있다 (Lavker et al. 1987; Choi and Oh 1997; Han et al. 1998). 그러나 이런 chemical에 비해 생체에 대한 부작용이나 세포독성이 상대적으로 적은 천연물을 이용하여 anti-skin aging drug을 개발하는 연구는 그 동안 부족했던 것이 현실이다. 한국은 오랜 역사를 통하여 화학물질이 아닌 다수의 천연물들을 임상에 적용하여온 역사를 가지고 있고 따라서 본 연구팀은 우리 남해안의 청정해역에서 자라는 곰피의 항피부 노화방지제로서의 개발 가능성에 주목하여 본 연구를 진행하였다. 연구 결과, 곰피추출물은 피부노화 및 skindarkness에 대한 대체치료물질 (alternative therapeutic agent)로서의 충분한 가능성을 보여주었다.

• 한국가축번식학회지
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• 제23권1호
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• pp.19-27
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• 1999

본 연구는 사람정자 추출물을 마우스 난모세포의 세포질내에 주입하여 단위발생에 미치는 영향을 구명하기 위하여 실시하였으며, 그 얻어진 결과는 다음과 같았다. 1. 마우스 난모세포가 0, 1.7 그리고 5mM의 칼슘농도를 가지는 PBS 배지로 각각 10 pl 씩 주입되었을 때, 전핵을 형성하고 제2극체를 방출하는 난모세포의 활성율은 각각 14.5, 9.8 그리고 14.9%였다. 칼슘농도별 난모세포의 활성율은 유의성이 인정되지 않았다 (P＞0.05). 2. 마우스난모세포에 가열하지 않은 사람정자 추출물 10 pl을 주입후 0, 1.7 그리고 5mM의 칼슘농도를 가지는 PBS 배지에서 각각 배양하였을 때, 1.7 mM 칼슘 농도에서의 난모세포 활성율(51.8%)은 0 과 5mM 칼슘 농도에서의 난모세포 활성율보다 유의하게 높았다. 3. 마우스 난모세포에 가열처리한 사랍정자 추출물 10 pl을 주입 후 0, 1.7 그리고 5mM의 칼슘농도를 가지는 PBS 배지에서 각각 배양하였을 때, 칼슘 농도별 난모세포 활성율은 유의성이 인정되지 않았다. 난모세포 활성율은 11.8∼17.0%의 범위에 있었다. 4. 마우스 난모세포에 가열하지 않은 사람 정자 추출물, 가열 처리한 사람 정자 추출물, 그리고 PBS 배지를 각각 l0 pl 씩 주입 후 1.7 mM 칼슘농도를 가지는 PBS 배지에서 각각 배양하였을 때, 가열하지 않은 사람 정자 추출물을 주입한 난모세포의 활성율이 54.5% 로 제일 높았다. 처리별 난모세포의 활성율은 유의성이 인정되었다(P＜0.05), 5. 마우스 난모세포에 가열하지 않은 l일령과 6일령 정자 추출물을 각각 l0 pl 씩 주입 후 1.7mM 칼슘농도를 가지는 PBS 배지에서 각각 배양하였을 때, 1일령 정자 추출물을 주입한 난모세포의 활성율은 60.0%로 6일령 정자 추출물을 주입한 난모세포의 활성율 11.1% 보다 유의하게 높았다. 이상의 결과를 종합해 보면 가열하지 않은 사람정자 추출물에는 oscillin 과 같은 난모세포 활성인자가 존재한다는 것이 입증되었다.

• 한국산업보건학회지
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• 제21권1호
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• pp.49-54
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• 2011

In order to develop the methods for exposure assessment and find susceptibility markers for the workers who are exposed to low doses of toluene, xylene and other chemical in petroleum industries, we investigated the application of P-450 expression in human lymphocytes utilizing mouse monoclonal anti-rat CYP2B1/2, the levels of toluene and xylene in air and their metabolite levels in urine with the levels of expressed CYP2B1/2 proteins. The general characteristics such as age, smoking and drinking habit were no significant difference between the control and exposed workers, but the working durations and working hours were significantly different. Workers in exposed group were exposed to the mean of 2.1 ppm (range, 0.00-4.54) of toluene and 0.3 ppm (rang, 0.00-1.23) of xylene. The mean concentration of urinary hippuric acid was low and less than 1/5 of the biological exposure index recommended by the Ministry of Employment and Labor Korea. Methyl hippuric acid in urine was not detected in control and exposed workers. Also, there were no significant differences in the levels of the urinary metabolites between the control and exposed group. When chemiluminescence dot blottings were carried out utilizing mouse monoclonal antibody against CYP2B1/2, the strong density dots corresponding to a mouse monoclonal antibody was observed in the human lymphocytes from the exposed workers. These results suggested that the chemiluminescence dot blot assay for CYP of lymphocytes should be valuable for identifying CYP expression as biomarkers in the workers exposed to toluene and xylene.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1791-1798
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• 2006

When ginsenoside Rg5, a main component isolated from red ginseng, was incubated with three human fecal microflora for 24 h, all specimens showed hydrolyzing activity: all specimens produced ginsenoside Rh3 as a main metabolite, but a minor metabolite $3{\beta},12{\beta}$-dihydroxydammar-21(22),24-diene (DD) was observed in two specimens. To evaluate the antiallergic effect of ginsenoside Rg5 and its metabolites, the inhibitory effect of ginsenoside Rg5 and its metabolite ginsenoside Rh3 against RBL-2H3 cell degranulation, mouse passive cutaneous anaphylaxis (PCA) reaction induced by the IgE-antigen complex, and mouse ear skin dermatitis induced by 12-O-tetradecanoilphorbol-13-acetate (TPA) were measured. Ginsenosides Rg5 and Rh3 potently inhibited degranulation of RBL-2H3 cells. These ginsenosides also inhibited mRNA expression of proinflammatory cytokines IL-6 and $TNF-{\alpha}$ in RBL-2H3 cells stimulated by IgE-antigen. Orally and intraperitoneally administered ginsenoside Rg3 and orally administered ginsenoside Rg5 to mice potently inhibited the PCA reaction induced by IgE-antigen complex. However, intraperitoneally administered ginsenoside Rg5 nearly did not inhibit the PCA reaction. These ginsenosides not only suppressed the swelling of mouse ears induced by TPA, but also inhibited mRNA expression of cyclooxygenase-2, $TNF-{\alpha}$, and IL-4 and activation of transcription factor NF-kB. These inhibitions of ginsenoside Rh3 were more potent than those of ginsenoside Rg5. These findings suggest that ginsenoside Rg5 may be metabolized in vivo to ginsenoside Rh3 by human intestinal microflora, and ginsenoside Rh3 may improve antiallergic diseases, such as rhinitis and dermatitis.

• Journal of Microbiology and Biotechnology
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• 제19권9호
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• pp.1070-1076
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• 2009

Telomerase reverse transcriptase (TERT), which prolongs the replicative life span of cells, is highly upregulated in 85-90% of human cancers, whereas most normal somatic tissues in humans express limited levels of the telomerase activity. Therefore, TERT has been a potential target for anticancer therapy. Recently, we described a new approach to human cancer gene therapy, which is based on the group I intron of Tetrahymena thermophila. This ribozyme can specifically mediate RNA replacement of human TERT (hTERT) transcript with a new transcript harboring anticancer activity through a trans-splicing reaction, resulting in selective regression of hTERT-positive cancer cells. However, to validate the therapeutic potential of the ribozyme in animal models, ribozymes targeting inherent transcripts of the animal should be developed. In this study, we developed a Tetrahymena-based trans-splicing ribozyme that can specifically target and replace the mouse TERT (mTERT) RNA. This ribozyme can trigger transgene activity not only also in mTERT-expressing cells but hTERT-positive cancer cells. Importantly, the ribozyme could selectively induce activity of the suicide gene, a herpes simplex virus thymidine kinase gene, in cancer cells expressing the TERT RNA and thereby specifically hamper the survival of these cells when treated with ganciclovir. The mTERT-targeting ribozyme will be useful for evaluation of the RNA replacement approach as a cancer gene therapeutic tool in the mouse model with syngeneic tumors.

• 대한화장품학회지
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• 제30권4호
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• pp.457-462
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• 2004

N-Acetyl-D-glucosamine (NAG)는 보습제로 사용되어지는 히아루론산의 구성물질인 뮤코 다당류의 일종이며, 특히 화장품으로서 응용은 보습제로서의 사용이 최초이다. 본 실험에서는 게나 새우의 껍데기에서 추출된 키틴을 탈아세틸화 하여 얻은 NAG를 화장품 원료로서 적용하고자 하였다. 현재 기능성 주름 원료로 알려진 레티놀(retinol)과 NAG를 비교하기 위하여 섬유아세포의 활성능력 및 콜라겐 생성촉진 효과를 비교 실험하였으며, 제형 내에서의 안정성을 위하여 HPLC로 역가를 측정하였다. 실험결과, 게의 껍질로부터 유도된 NAG는 피부에 자극을 전혀 주지 않으면서 섬유아세포의 세포활성 및 콜라겐의 생성을 촉진시키는 효과를 나타내었으며, 헤어리스 마우스를 대상으로 실험을 실시한 결과 피부층의 변화를 통하여 주름의 감소 효능을 볼 수 있었다.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.321-340
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• 1995

The regeneration of destructed periodontal tissues is one of the ultimate objectives of periodontal therapy. Guided tissue regeneration technique was developed for the ideal regeneration of periodontal tissues. In order to investigate the role of fibronectin, laminin and tenascin in the regenerating process of periodontal tissues, the expanded PTFE barrier membranes(Gore Associates, USA) removed from the patients who had been treated by guided tissue regeneration(GTR) and guided bone regeneration(GBR) techniques were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, and immunohistochemically processed by Avidin-Biotin peroxidase complex method for detecting fibronectin, laminin and tenascin. Monoclonal mouse anti-human fibronectin antibody(Oncogene Science, USA., 1:100), monoclonal mouse anti-human laminin antibody(Oncogene Science, USA., 1:50) and mouse anti-human tenascin antibody(Oncogene Science, USA, 1:10) were used as primary antibodies. The light microscopic findings were as follows: (1) The distribution of fibronectin, laminin and tenascin was various according to the area of barrier membranes. (2) The distribution of fibronectin in case of GBR was extensive in the tissue on the outer surface of barrier membranes, and rare in the intervening space and on the inner surface. In case of GTR it was extensive on the outer surface and in the intervening space, and rare on the inner surface. (3) The distribution of laminin was rare in the tissue on the outer, the inner surface and intervening space of barrier membranes, regardless of GBR or GTR. (4) In case 'of GBR rare distribution of tenascin was observed on the outer surface only, except the inner surface and the intervening space of barrier membranes. In case of GTR the distribution of tenascin was extensive in the tissue on the outer surface, rare in intervening space and the inner surface. The results suggest that fibronectin, laminin and tenascin may play a important role in the regenerating process of periodontal tissue, and they may affect the outcome of healing.

• 대한수의학회지
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• 제34권4호
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• pp.769-779
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• 1994

The root of Bupleurum falcatum L.(BF) has been widely used in oriental medicine as a major camponent in many prescriptions for chronic hepatitis, renal disease, tuberculosis and some other infectious diseases. Many attempts have done to investigate the therapeutic effects of these principles. However, any kinds of screenig on immune regulatory- and antitumor- effects of BF has not been reported. The present study, therefore, was undertaken to investigate the BF-effects on cellular- and humoral-immune responses, phagocytic activities of macrophages, lymphokine- and Immunoglobulin(Ig)-production of lymphocytes, tumorigenesis of implanted sarcoma 180 cells and B16 melanoma cells, and proliferations of some tumor cell lines(Fsa II, 3LL and EL4). BF increased phagocytic activities of mouse peritoneal macrophages in a dose- and time-dependent fashion. Arthus reaction and antibody responses to SRBC were slightly enhanced but delayed hypersensitivity was depresed when BF was injected before- and after-SRBC sensitization. BF inhibited the proliferative responses of human tonsillar lymphocytes to PHA- and Con A-stimulation but slightly augmented the response of these cells to Staphylococcus aureus Cowan 1(SAC)-activation. Ig secretion of human mononuclear cells activated with SAC was slightly increased by BF. BF significantly augmented the SAC-induced IL 6 production of human mononuclear cells but not influenced Con Ainduced IL 2 secretion. NK cell activities of mouse splenocytes were somewhat increased when BF was pretreated and this responses were due to the increment of binding affinities of effector cells to target cells and of lytic activities of effector cells against target cells. In vitro BF significantly inhibited the proliferations of cancer cells such as Fsa II, 3LL and EL4 strains. BF decreased not only the frequency of tumor induction but also the tumor size per sarcoma 180 or B16 cell-implanted mouse. Taken together, these results indicate that BF is one of the potential immunomodulator, and suggest its possibility to be used as a desirable antitumor agent.

• Journal of Periodontal and Implant Science
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• 제23권3호
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• pp.605-621
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• 1993

The gingival hyperplasia refers to an increase in the size of the gingival tissue produced by an increase in the number of its component cells. In order to investigate the cellular change in epithelium and subepithelial tissue of noninflammatory gingival hyperplasia, the gingival tissues were surgically obtained from the patients with dilantin gingival hyperplasia and idiopathic gingival hyperplasia. The excised tissue samples were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, mounted on glass slides coated with 3-aminopropyltriethoxysilane(Sigma Chemical Co., St. Louis, MO, U.S.A.) and immunocytochemically processed by Avidin-Biotin peroxidase complex method for detecting proliferating cell nuclear antigen, tenascin and collagen type IV. Monoclonal mouse anti-human PCNA antibody(Oncogene Science, Uniondale, NY, U.S.A., 1 : 250,000), monoclonal mouse anti-human tenascin antibody(Chemicon-International Inc., Temecula, CA, U.S.A., 1:5,000), and monoclonal mouse anti-human collagen type IV(Dakopatts, Glostrup, Denmark, 1: 50) were used as primary antibodies. The results were as follows: 1. In non-inflammatory gingival hyperplasia, the positive reaction to proliferating cell nuclear antigen was localized in the basal cell layer of gingival epithelium and well-developed rete pegs. 2. The positive reaction to tenascin was shown in the connective tissue subjacent to basament membrane of gingival tissue, and especially strong positive reaction was noted in the tip portion of connective tissue projections. 3. The positive reaction to collagen type IV was localized along the basement membranes of gingival epithelium and blood vessels. The results suggest that connective tissue enlargement may affect the proliferation of gingival epithelium.