• Clinical and Experimental Reproductive Medicine
• /
• 제21권2호
• /
• pp.183-190
• /
• 1994

Purpose of the present study was to find the optimal culture conditions for the maturation and fertilization of immature oocytes by the use human body fluids and gonadotropins (Gn) in the mouse model. Cumulus-enclosed mouse immature oocytes were incubated in the medium containing various human body fluids with or without Gn in vitro, and examined to confirm nuclear maturation (NM) and fertilization. Female ICR mice were stimulated with 7.5 IU pregnant mares' serum gonadotropin (PMSG). Cumulus-enclosed immature oocytes were isolated at 48-52 hr post PMSG injection and cultured in TCM 199 supplemented with various concentrations (20, 50, and 70%) of human body fluids such as fetal cord serum (hCS), follicular fluid (hFF), peritoneal fluid (hPF) and amniotic fluid (hAF) in the presence or absence of 10 IU/ml PMSG and 10 IU/ml human chorionic gonadotropin (hCG) for 18 hr. Fetal calf serum (FCS) was used as a control for the supplements. Matured oocytes were fertilized with sperm collected from the epididymis of male mice. Fertilization was conducted in T6 medium containing 15 mgl ml bovine serum albumin, and confirmed at 6 hr post-insemination. Evaluation of nucler maturation and fertilization was carried out by rapid staining using fuchin. There was no significant difference between the effects of human body fluids and FCS supplements on nuclear maturation of cumulus enclosed mouse immature oocytes. When maturation medium was supplemented with 20% hPF or 20% hAF, fertilization rates were significantly (P<0.01) lower than that of 20% FCS, hCS and hFF groups. However, higher concentrations of body fluids during IVM were not more beneficial on fertilizability of oocytes. The addition of Gn significantly increased the fertilization rates in hPF and hAF groups (hPF without Gn; 51.5%, compared with 85.1% for addition of Gn, and hAF without Gn; 30.1% compared with 85.8% for addition of Gn) at 20% concentration. These results suggest that human body fluids at 20% concentration and gonadotropins can be used as supplements for the maturation of mouse immature oocytes in vitro. When gonadotropins supplemented with the human body fluids in the maturation medium, fertilizability of mouse immature oocytes was increased in hPF and hAF groups. These results can be applied to maturation of human immature oocytes in vitro.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1996년도 제4회 추계심포지움
• /
• pp.55-64
• /
• 1996

mouse chromesome2에 있는 agouti locus는 정상적으로는 털색깔을 조절하는 gene이다. mouse agouti gene은 최근에 cloning 되었고 131 amino acid peptide와 consensus signal peptide를 encode한다고 보고되었다. 이 논문에서 interspecies-DNA hybridization approach를 이용하여 mouse agouti gene의 human homologue를 cloning 하였다. Sequence analysis 결과, 이는 mouse gene에 85% 유사하였고 consensus signal peptide sequence 를 포함하는 132 amino acid를 coding하였다. somatic-cell hybrid mapping pannel과 Fluorescence-in-situ hybridization에 의한 chromosomal mapping을 한 결과, agouti gene은 MODY (maturity onset diabetes of the young), myeloid leukemia locus 등이 위치한 human chromosome 20q 11.2에 mapping 되었다. 성인 tissue로부터 추출한 RNA를 이용한 발현연구에 의하면 human agouti gene은 adipose tissue와 teatis에 발현되었다.

• 생약학회지
• /
• 제26권3호
• /
• pp.253-258
• /
• 1995

The purpose of this research was to investigate the effects of petroleum ether extract of Euonymus alatus (EAP) on the proliferation of human tumor cells. EAP inhibited the proliferation of HeLa, Hep G2, KHOS/NP and A431 cells. The cytotoxicity of doxorubicin on human tumor cells and Balb/c 3T3 cells were increased by the combination of EAP. EAP did not affect the proliferation of Balb/c 3T3 cells, mouse spleen cells and human lymphocytes. These results suggest that EAP has the cytotoxicity on human tumor cells without cytotoxicity on Balb/c 3T3 cells, mouse spleen cells and human lymphocytes, and increase the cytotoxicity of doxorubicin.

• 산업공학
• /
• 제17권1호
• /
• pp.113-120
• /
• 2004

The increased use of computers has introduced a variety kind of human-computer interfaces. Mouse is one of the useful interface tools to place the cursor on the desired position on the monitor. This paper suggested a foot controlled isotonic mouse which was similar to the ordinary hand-controlled mouse except that positioning was controlled by the right foot and the clicking was performed by the left foot. Experimental results showed that both the index of difficulty(IOD) and the display-control gain(DC gain) varied the total movement time in a target acquisition task on the monitor. The present authors also drew the optimal display-control gain of the foot-controlled isotonic mouse over the index of difficulty of 1.0 to 3.0. The optimal display-control gain, i. e., 0.256, could be used when designing a foot-controlled isotonic mouse.

• Clinical and Experimental Reproductive Medicine
• /
• 제11권2호
• /
• pp.51-57
• /
• 1984

The success of human in vitro fertilization (IVF) & embryo transfer (ET) has focused attention on the culture conditions that can provide optimal development of the preimplantation embryo. Studies of in vitro fertilization using mouse have direct implications to human IVF, since similar conditions are used for both species. Mouse IVF as a quality control system for human IVF & ET was studied since Feb., 1984. The results were as follows: 1. Egg retrieval following superovulation in IeR mice was l5.1${\pm}$5.3 eggs ovulated/mouse (Mean${\pm}$ S.D.) 2. In vitro cleavage rate was 61.7% (1146 eggs cleaved/l858 eggs inseminated) and % blastocyst was 42.6%. 3. In comparison with two media of Ham's F-10 and m-KRB, in vitro cleavage rate were 40.9%/63.l% and %blastocyst were 44.3%/61.2% (P<0.05). 4. It was concluded that mouse IVF system has a valuable place in human IVF & ET as a quality-control system and in human reproductive physiology as a research model.

• 약학회지
• /
• 제38권3호
• /
• pp.311-321
• /
• 1994

Anticancer effects of Aloe on sarcoma 180 in ICR mouse or human cancer cells were determined. Sarcoma 180 cells were inoculated subcutaneously into male ICR mouse to determine effect of Aloe on tumor gowth, or inoculated intraperitoneally into male ICR mouse to determine effect of Aloe on life span prolongation, followed by oral administration of Aloe vera(10 mg/kg/day, 50 mg/kg/day) or Aloe arborescens(10 mg/kg/day, 100 mg/kg/day) once a day for 14 days. The administration of Aloe vera or Aloe arborescens did not suppress tumor growh. However the life span of ICR mouse was prolonged to 19%(p<0.05), 22%(p<0.05) and 32%(p<0.05) by administration of Aloe vera 10 mg/kg/day, Aloe vera 50 mg/kg/day, and Aloe arborescens 100 mg/kg/day, respectively. To determine anticancer effect of Aloe in vitro, Aloe extract was added to the culture of human gastric cancer cells(SNU-1) and colorectal cancer cells(SNU-C2A), and concentration of Aloe to inhibit cancer cell growth was determined using MTT(3-[ 4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) cytotoxicity assay. High $ID_{50}$ values of Aloe vera and Aloe arborescens against gastric cancer cell line(SNU-1) and colorectal cancer cell line(SNU-C2A) suggest that Aloe gel does not have anticancer effect on these specific human cancer cells although high concentration of Aloe inhibited growth of human cancer cells significantly.

• Genomics & Informatics
• /
• 제6권1호
• /
• pp.32-35
• /
• 2008

Little evidence supports the existence of imprinted genes in chicken. Imprinted genes are thought to be intimately connected with the acquisition of parental resources in mammals; thus, the predicted lack of this type of gene in chicken is not surprising, given that they leave their offspring to their own heritance after conception. In this study, we identified several imprinted genes and their orthologs in human, mouse, and zebrafish, including 30 previously identified human and mouse imprinted genes. Next, using the HomoloGene database, we identified six orthologous genes in human, mouse, and chicken; however, no orthologs were identified for SLC22A18, and mouse Ppp1r9a was not included in the HomoloGene database. Thus, from our analysis, four candidate chicken imprinted genes (IGF2, UBE3A, PHLDA2, and GRB10) were identified. To expand our analysis, zebrafish was included, but no probe ID for UBE3A exists in this species. Thus, ultimately, three candidate imprinted genes (IGF2, PHLDA2, and GRB10) in chicken were identified. GRB10 was not significant in chicken and zebrafish based on the Wilcoxon-Mann-Whitney test, whereas a weak correlation between PHLDA2 in chicken and human was identified from the Spearman's rank correlation coefficient. Significant associations between human, mouse, chicken, and zebrafish were found for IGF2 and GRB10 using the Friedman's test. Based on our results, IGF2, PHLDA2, and GRB10 are candidate imprinted genes in chicken. Importantly, the strongest candidate was PHLDA2.

• 한국가축번식학회지
• /
• 제18권1호
• /
• pp.71-81
• /
• 1994

These experiments were carried out to investigate the effects of human follicular fluid (hFF) as a protein supplement on development of mammalian embryo as well as to find out ways toward effective use of hFF. The developmental rates of mouse embryos to the blastocyst and implantation stages were significantly higher in T6 ＋hFF than T6＋hFCS. Classified hFF according to the maturity of contained oocytes (M-hFF and Im-hFF), and compared the rates of development of mouse embryo cultured in M-hFF or Im-hFF to culture medium T6. Total protein, albumin and estradiol concentrations were higher in M-hFF than Im-hFF (P<0.05). The developmental rates of mouse embryos to the blastocyst and hatching blastocyst stages cultured in Im-hFF were significantly lower than those in M-hFF and the basic medium. In accordance of the results of human IVF, hFF has been divided into 4 groups. The developmental rates of mouse embryos to the blastocyst stage in presense of hFF from pregnant patients, who have good grade embryos, were significantly higher than those in hFF from patients who have poor grade embryos or were not pregnant. In addition, the rates of development of human embryo were compared in presense of BSA, hFF or hFCS. The developmental rates of human embryos cultured in Ham's F10＋hFF were significantly higher than those in the Ham's F10＋BSA. These results suggests that the culture system using hFF could improve the development ability of mammalian embryos and the viability of blastocysts cultured in vitro.

• 한국가축번식학회지
• /
• 제27권1호
• /
• pp.15-23
• /
• 2003

본 연구는 VSV-G glycoprotein을 envelope으로 하는 pseudotyped retrovirus vector system을 이용하여 쥐의 유방상피세포인 HC11에서 human Lactadherin 유전자의 발현을 확인하고자 하였다. 실험에 사용한 vector는 개체내에서의 외래 유전자의 지속적인 발현에 의한 생리적인 부작용을 최소화하기 위한 구조로, 조직특이적이며 lactogenic hormone에 의해 유도적인 활성을 가지는 것으로 알려진 WAP promoter의 통제하에 도입하고자 하는 외래 유전자를 위치하도록 하였다. WAP promoter의 대조군으로 지속적인 활성을 나타내는 $\beta$-actin promoter를 사용하였으며, 이 각각의 promoter와 marker gene으로 E. coli LacZ gene을 재조합한 후 retrovirus vector system을 이용하여 HCll에 도입하였다. 세포의 genome 내로의 유전자의 전이는 PCR을 통해 확인하였고, RT-PCR의 수행으로 유전자의 발현을 확인하였다. Lactadherin 유전자를 이용한 실험도 동일한 과정으로 수행하였으며, RT-PCR의 결과에서 HCll 세포에서 Lactadherin 유전자의 발현이 insulin을 단독으로 처리한 군에 비해 insulin, hydrocortisone, prolactin을 동시에 처리한 군에서 우월하게 나타나는 것으로 확인되었다. 그러나 insulin 단독 처리군에서 유전자의 발현이 약하게 나타나는 것으로 관찰되어 WAP promoter의 leakiness에 대한 재고의 필요성이 요구되었다.

• Clinical and Experimental Reproductive Medicine
• /
• 제44권3호
• /
• pp.119-125
• /
• 2017

Objective: In vitro culture of preimplantation embryos is improved by grouping embryos together in a drop of media. Individually cultured embryos are deprived of paracrine factors; with this in mind, we investigated whether the addition of a single embryo-secreted factor, interleukin-6 (IL-6), could improve the development of individually cultured embryos. Methods: Mouse embryos were cultured individually in $2{\mu}L$ of G1/G2 media in 5% oxygen and supplemented with a range of doses of recombinant mouse or human IL-6. Results: Mouse IL-6 increased hatching at doses of 0.01 and 10 ng/mL compared to the control (93% and 93% vs. 78%, p< 0.05) and increased the total number of cells at a dose of 0.1 ng/mL compared to the control ($101.95{\pm}3.36$ vs. $91.31{\pm}3.33$, p< 0.05). In contrast, the highest dose of 100 ng/mL reduced the total number of cells ($79.86{\pm}3.29$, p< 0.05). Supplementation with human IL-6 had a different effect, with no change in hatching or total cell numbers, but an increase in the percentage of inner cell mass per embryo at doses of 0.1, 1, and 100 ng/mL compared to the control ($22.9%{\pm}1.1%$, $23.3%{\pm}1.1%$, and $23.1%{\pm}1.1%$ vs. $19.5%{\pm}1.0%$, p< 0.05). Conclusion: These data show that IL-6 improved mouse embryo development when cultured individually in complex media; however, an excess of IL-6 may be detrimental. Additionally, these data indicate that there is some cross-species benefit of human IL-6 for mouse embryos, but possibly through a different mechanism than for mouse IL-6.