• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.13-20
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• 1997

The objective of this study was to determine the effect of EGF on the preimplantation development of mouse IVF embryos and their ICM and TE cell number. And also, we examined the expression of EGF-R protein on embryonic development by indirect immunofluorescence. The results obtained in these experiments were summarized as follows; Group culture (5 embryos/ 25 ${\mu}l$) showed more improved development rate to blastocyst than singly culture. This inferior development of singly cultured 2-cell embryos improved by the addition of EGF. Especially, 2-cell embryos cultured singly in 10 ng/ml of EGF (62.4%) indicated significant difference in development to blastocyst compared with control group (47.9%). Also, cell number of ICM and TE by differential labelling showed the increased pattern in the EGF treatment group. The stimulating effect of EGF with the development level was significantly increased after 4-cell stage (p<0.05). ICM proportion also showed the increased pattern with the developmental level in the EGF treatment group. In addition, expression of EGF-R by indirect immunofluorescence detected after 4-cell stage. Therefore, EGF could stimulate preimplantation mouse embryo development by binding with expressed EGE-R after 4-cell stage and produce the more increased ICM and TE cell number of blastocyst.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.279-288
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• 1996

The objective of this study was to determine the effect of EGF on the development of IVM/IVF bovine embryos and their ICM and TE cell number. In addition, we examined the combined effect of EGF and coculture to the bovine embryo development and the expression of EGF-R protein on bovine embryos by indirect immunofluorescence. The results obtained in these experiments were summarized as follows: When the IVM/IVF 4- to 8-cell embryos were treated at 0, 1, 10, 100 ng/ml of EGF, EGF treatment group showed improved development to blastocyst and increased pattern of ICM and TE cell number compared with control, although there is not significantly different. The stimulating effect of EGF (10 ng/ml) to the develop ment level of IVM/IVF bovine embryos significantly increased development rate to blastocyst after 8-cell stage (p<0.05), although there is no significant effect to the increase of ICM and TE cell numbers. Also, expression of EGF-R on the bovine embryonic stage by indirect immunofluor escence presents after 4-cell stage and the intensity of the EGF-R staining was variable with the development progression. On the other hand, embryos cultured in coculture group added either with or without EGF commonly indicated the significant difference in development rate to blastocyst and Total cell number compared with control. These results suggest that the a addition of EGF to the coculture may stimulate the coculture effect between IVM/IVF bovineembryos and cumulus cells. Therefore, EGF could promote preimplantation bovine embryo development by binding with expressed EGF~R after 4-cell stage, and stimulate the production of embryotrophic factors from the coculture environment. Also, the present study showed that there was no significant effect of EGF to the increase of ICM and TE cell number although the rate of blastocyst significantly increased when treated with EGF after 8-cell stage (p<0.05).

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.21-26
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• 1997

This study was undertaken to examine the expression of EGF-R protein on ICM obtained from mouse IVF/IVC blastocyst by immunosurgery and indirect immunofluorescence(IIF). ICM cells used for this experiment were obtained from immunosurgery of mouse blatstocysts produced at 96 h after IVF, and recovered ICMs were assayed for cell viability and expression of EGF-R. The results obtained in this experiment were summarized as follows: when blastocysts were exposed to rabbit anti-mouse serum (antiserum) for 15-30 min. and then transferred them to guinea pig serum (complement) for 15-60 min., recovery rates of isolated ICMs were 8.0-84.2%. Especially, the best recovery (84.2%) of ICMs was obtained when exposure time to antiserum and complement was 30 min. and 60 min., respectively. In addition, when viability of isolated ICMs after immunosurgery was assessed by live/dead staining method, in all groups viability (93.8-100.0%) of isolated ICMs were not damaged if separated from TE cell. Also, we detected the expression of EGF-R on ICM cell by IIF. Therefore, these results suggest that EGF-R expression on the ICMs can stimulate the higher usablity of exogenous EGF to improve the preimplantation embryo development in vitro.

• Journal of Chest Surgery
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• v.30 no.9
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• pp.854-861
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• 1997

CYPRA 21-1 is known to be a cytokeratin 19 fragment, and it can be detected by using two specific monoclonal antibodies (KS 19-1 and BM 19-21) and can be clinically applied as a useful circulating tumor marker The epidermal growth factor receptor (EGF-R) expression was evaluated and characterized by its tyrosine protein kinase activity and by its ligand-stimulated autophosphorylation, a property shared with other peptide growth factor receptors. Autocrine or para'urine action was initiated by a growth factor, or by a transforming growth factor o, which had an extensive homology with EGP and which also stimulated tyrosine kinase activity on the EGF-R. The CYFRA 21-1 and the EGF-R levels in 30 patients with primary lung tumors were investigated. There were 24 patients with squamous cell carcinomas and 6 patients with adenocarcinomas. Specimen 5 mm3 in size were sampled at three different locations ; the main lesion, the boundary between the lesion and the unaffected tissue, and the unaffected tissue of the patients. The results were as follows 1. The CYPRA 21-1 concentration in the cancer boundary, the most malignant region,(348.6 : 89.9 ng/ml) was the lowest value. The CYFRA 21-1 concentration in unaffected tissue,(718.4$\pm$77.8 ng/ml) was higher than that in the main lesion. which had intact cellularity. 2. The EGF-R concentration in the main lesion was higher than that in the unaffected tissue, and the EGF-R concentration in a squamous cell cacinoma was higher than that in an adenocarcinoma. also, the EGF-R concentration in the cancer b undary was highest at stage 1, ll. The EGF-R concentration was higher in the main cancer lesion that in the unaffected tissue at stage 111, IV. 3. The CYFRA 21-1 was a cytoplasmic skeleton and the EGF-R was a cell-wall component; there was no correlation. In conclusion, CYFRA 21-1 was abundant in the cytoplasm but had a higher concentration in the unaffected tissue than in the main cancer lesion. The CYFRA 21-1 concentration of the tissue did not reflect the amount of cancer activity, the EGP-R was located in the cell membrane, the level of tissue that reflects cancer activity, so the main cancer lesion had a higher concentration than the unaffected tissue. CYFRA 21-1 is not a useful tumor maker at the tissue level. Because the EGF-R concentration re(looted the cancer activity, its a useful tumor marker for lung cancer.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.39-46
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• 1997

본 연구는 체외생산된 소 배반포기배의 inner cell mass (ICM) 세포에서 epidermal growth factor-receptor (EGF-R)의 발현 유무를 immunosurgery와 indirect immunofluorescence (간접 면역 형광방법)을 이용하여 조사하고자 실시하였다. 본 실험에 사용된 ICM 세포는 체외수정 후 7∼8일째에 회수된 소 배반포기배로부터 immunosurgery 방법을 실시하여 얻어졌으며, 회수된 ICM세포는 live/dead 염색방법을 통한 생사 유무와 EGF-R 발현 유무 조사에 공시되었다. 특히, 배반포기배에 대한 immunosurgery를 위해 trophectoderm 세포에 대한 rabbit anti-bovine trophectoderm cell antibody (RABTE)를 제조하여 사용하였다. 결과를 요약하면 다음과 같다. ICM세포의 회수율은 RABTE와 guinea pig serum (complement)에 각각 15∼30분과 15∼60분동안 처리했을 경우 16.7∼74.2%였으며, 또한 처리시간이 각각 30분과 30분일 때 가장 높은 회수율(74.2%)을 얻었다. Immunosurgery 후 얻어진 ICM세포의 생존 유무를 조사하기 위해 live/dead 염색 방법을 이용하였던바, ICM세포의 생존율은 complement가 60분 처리된 군(69.3%)을 제외한 모든 처리군에서 84.0∼91.6%의 높은 생존율을 나타냈다. 또한, 회수된 ICM세포에 대한 EGF-R의 존재를 확인하였다. 따라서, ICM세포에서의 EGF-R의 발현은 인위적으로 첨가된 EGF의 이용 가능성을 높임으로서 체외에서의 착상전 배 발달을 증진시킬 수 있을 것으로 사료된다.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.2
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• pp.175-184
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• 2019

The epidermal growth factor (EGF) has a intrinsic function of inducing growth and proliferation of cells through interacting with cell membrane receptors in human epidermis and dermis layer. These functions of EGF are used as a main ingredient for wound healing medicines and anti-aging cosmetics. As a cosmetic ingredient, the EGF has a problem in exhibiting its natural efficacy due to the lack of the ability to penetrate through the stratum corneum, which is known as the skin barrier. In this study, a recombinant human epidermal growth factor ($MTD_{151}-EGF$) fused with the macromolecule transduction domain $(MTD)_{151}$ with the skin penetration ability was developed to improve the skin penetration efficiency of the EGF. Expression of $MTD_{151}-EGF$ was performed in E. coli transformed with a vector encoding the $MTD_{151}-EGF$ gene and then purified. The purified $MTD_{151}-EGF$ was evaluated using cell proliferation assay, cytotoxicity test and skin penetration test by franz diffusion cell assay and artificial skin. Cell proliferation activity of $MTD_{151}-EGF$ purified to high purity of 99% or above was equivalent to the EGF or better, and cytotoxicity was not observed. In addition, the $MTD_{151}-EGF$ showed an excellent penetration efficiency compared to the EGF in the skin penetration test with EGF and $MTD_{151}-EGF$ labeled by FITC in an artificial skin penetration model. Based on the quantitative analysis of the penetrating substance using franz diffusion cell assay, the amount of penetration was about 16 times more than that of EGF. These results can be regarded as an effective alternative to improve the existing physical transdermal penetration method related to the use of various active ingredients for cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.25 no.1
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• pp.107-119
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• 1999

3 Kinds of PEG-hEGF conjugate, PEG5-hEGF, PEG5-hEGF, PEG10-hEGF, were synthesized. Major fractions containing 2 chains of PEG were separated by preparative GPC. Molecular weight was estimated by GPC-MALLS, TOF-Mass and SDS-PAGE, and the data were well corresponding to calculated one. The cell proliferation effect of the conjugates was evaluated, Indicating that the conjugate bound to longer chain PEG exhibits lower activity. Selective modification of hEGF and activity preservation of PEG-hEGF conjugate are undergoing.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.2
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• pp.340-347
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• 1994

The specimens used in this study were obtained from patients with primary gastric carcinoma and adjacent non-malignant mucosa from the same patients. Using the techniques of immunocyto chemicstry and in situ hybridization, transforming growth $factor-{\alpha}(TGF-{\alpha})$ and epiderimal growth factor receptor (EGF-R) nRNAs were identified. $TGF-{\alpha}$ was observed in macrophages and dividing tumor cells but, not in normal cells. EGF-R was observed both in malignant and non-malignant gastric tissues. Although normally, $TGF-{\alpha}$ is not seen in normal gastric tissues, $TGF-{\alpha}$ was discovered in the adjacent non-malignant tissue of histolgically normal, which strongly suggest that $TGF-{\alpha}$ is involved in the differentiation of cancer cells. Immunocytochemicstry using EMB-11 antibody identified the existence of macrophages which express $TGF-{\alpha}$ and EGF-R mRNA. Protein products of EGF-R was identfified using monoclonal antibody. Cancer cells were also identified in the non-malignant normal tissues by the method of immunocytochemicstry using carcino embryonic antigen (CEA)antibody. It is considered that the activity of $TGF-{\alpha}$ increased as tumor cell prolifierates. Immunocytochemistry and in situ hybridization techniques can be used to diagnose gastric cancer along with the use of ${\alpha}-feto$ protein and CEA.

• Journal of Embryo Transfer
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• v.19 no.2
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• pp.173-183
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• 2004

This study was carried out to examine the effects of epidermal growth factor (EGF) and the number of cumulus-oocyte complexes (COCs) on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). COCs were collected from antral follicles of porcine ovaries collected from abattoir, and were maturated in modified NCSU-23 (mNCSU-23) with 10% pFF, 0.6 mM cysteine, 50 ${\mu}mM{\beta}-mercaptoethanol$, 1 mM dbcAMP, 10 IU/mL PMSG and 10 IU/mL hCG, which was supplemented with or without 10 ng/mL EGF and into which 50 or 15 COCs per droplet was put. Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium (mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU- 23. The results are as follows. 1.In the result of IVM, 10 ng/mL EGF supplement duplicated the percentage of C4 group of COCs(41% vs 81%). But the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, and there was not a significant interaction between the two factors, either. 2. In the result of IVF, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, or was not a significant interaction between the two factors, in the rate of sperm penetration, in the percentage of oocytes with male pronucleus (MPN), and in the rate of polyspermy. 3. In the result of IVD, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet in the percentage of cleaved oocytes. There was not significantly different between the number of COCs per culture droplet, but between control and EGF supplemented (p<0.01) in the percentage of blastocysts, the number of inner cell mass (ICM), trophectoderm (TC) and total cells. There was no significant interaction between the two factors anywhere. These results suggested that 10 ng/mL EGF supplement into mNCSU-23 for IVM was effective in the production of more as well as better blastocysts during IVD through increasing the number of cells in those.

• Archives of Pharmacal Research
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• v.26 no.8
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• pp.659-665
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• 2003

We have studied the effect of rhEGF on the buccal mucosal ulcer healing. rhEGF was rapidly degraded upon incubation with the hamster buccal mucosal homogenates; The degradation of rhEGF was significantly inhibited by sodium lauryl sulfate (SLS). Eudispert hv hydrogel and Polycarbophil 974P hydrogel were prepared for rhEGF delivery and their mucoadhesiveness was measured by the $Instron^R$ method. The mucoadhesive force of Eudispert hv was significantly greater than that of Polycarbophil 974P. Moreover, rhEGF in Eudispert hv hydrogel remained stable for about 2 months. To evaluate the ulcer healing effect of rhEGF, the buccal mucosal ulcer was induced in golden hamsters using acetic acid. At 24 h after administration of rhEGF/Eudispert hv hydrogel, the ulcerous area was decreased compared with rhEGF solution and, as a result, the curative ratio was $36.8\pm5.68$%. By the addition of SLS (0.5%) to Eudispert hv hydrogel, the curative ratio increased 1.5 times. The mechanism of the action was probably due to a combination of protection of the drug against proteases present in mucosa and prolongation of the release of rhEGF from the formulation at the site of action.