• Title/Summary/Keyword: human cell culture

Search Result 1,026, Processing Time 0.033 seconds

Genetic Characterization for Human Enterovirus Isolated from Busan Region in Korea (부산지역에서 분리된 엔테로바이러스의 유행양상 분석 및 유전자형 연구)

  • Kim, Nam-Ho;Min, Sang-Kee;Park, Eun-Hee;Park, Yon-Koung;Kwan, Sun-Mok;Jin, Seong-Hyeon
    • Journal of Life Science
    • /
    • v.20 no.6
    • /
    • pp.907-913
    • /
    • 2010
  • Human enteroviruses (HEV) are considered one of the major infectious causes of central nervous system infections such as aseptic encephalomeningitis in pediatrics. This study was focused on providing information related to genetic characteristics and diversities of HEV which prevailed between 2007 and 2009 in Busan, Korea. A total of 2,743 specimens were collected from children and screened for isolation of HEV by cell culture and RT-PCR. Among the specimins, 240 isolates were grouped into 21 different HEV serotypes using VP1 RT-PCR. The major etiological agents were CV-A6 and CV-B2 in 2007, E-6 and E-30 in 2008 and CV-B1 in 2009. The occurrence of HEV infections was the most frequent in the summer (May to August, 188 cases, 78.3%). Most of the isolates were identified from specimens from children under 10 years old, with the highest occurrence in the 2 to 4 year old range (15.2%). However, there were no significant differences between male and female children for the isolates. For analyzing genetic characterization, VP1 gene was amplified by RT-PCR and sequenced. The phylogenetic tree was established by Clustal W method using DNASTAR software. Using the sequence analysis of the VP1 region, it was classified into 2 groups; HEV-A and HEV-B. The HEV-A group contained 6 serotypes and sequences of 31 isolates were compared within each serotype. The HEV-B group contained 10 serotypes and the sequences of 41 isolates were compared within each serotype. Homology analysis of the VP1 region showed that the identity scores of HEV-A and B isolates were different. In conclusion, genetic divergences were observed among the isolates from children between 2007 and 2009 in Busan.

Fermented Jeju Soybean Extract Promotes Hair Growth in Human Hair Follicle Organ Culture and Clinical Trial (모낭기관배양과 임상에서 제주푸른콩발효추출물의 육모 효능에 미치는 영향)

  • Lee, Yonghee;Shin, Seung Hyun;Kim, Sehyun;Jeong, Gyusang;Hong, Min Jung;Park, Hyeokgon;Kim, Seung Hun;Kim, Eun Joo;Kang, Young-Gyu;Park, Byung Cheol;Park, Won Seok;Kim, Su Na;Kim, Hyoung-June
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.47 no.3
    • /
    • pp.255-263
    • /
    • 2021
  • Soybean extract is known to play an important role in preventing and treating diseases associated with aging, cancer, obesity, and cardiovascular disease. A recent has revealed that soybean extract has a potent effect on hair growth in in vitro, in vivo, and clinical studies. Recently, it has been reported that their fermented extracts exhibit numerous and high efficacy, as compared to general extracts. However, the underlying mechanisms that induce hair growth after using fermented soybean extract are not well understood. The present study aimed to determine the effects of fermented Jeju soybean (FJS) extract on hair growth, with a focus on the underlying mechanisms similar to those of minoxidil. We conducted in vitro and ex vivo investigations and clinical studies. FJS extract enhanced dermal papilla cell proliferation, VEGF levels, and potassium channel opening. Moreover, it promoted human hair follicle elongation. These effects were comprehensively demonstrated in the clinical results, in which FJS extract-containing shampoo improved hair density after 24 weeks of utilization. Collectively, the results of this study demonstrate that FJS extract promotes hair growth and inhibits hair loss through a mechanism similar to that of minoxidil in hair follicles.

Identification of CM1 as a Pathogenic Factor in Inflammatory Diseases and Cancer

  • Bae, Se-Yeon;Kim, Hyem-In;Yu, Yeon-Sil;Lee, Na-Eun;Kong, Joo-Myoung;Kim, Hang-Rae;Hwang, Young-Il;Song, Yeong-Wook;Kang, Jae-Seung;Lee, Wang-Jae
    • IMMUNE NETWORK
    • /
    • v.11 no.3
    • /
    • pp.175-181
    • /
    • 2011
  • Background: CM1 (centrocyte/-blast marker 1) was defined by a mAb against concavabalin-A (ConA) activated PBMC. It is expressed in germinal center of human tonsil and on the surface of activated PBMC as well as cancer cells. Recently, increased productions of pro-inflammatory mediators were detected from activated PBMC by CM1 ligation. Methods: However, there is a limitation to explain the exact role of CM1 on inflammation and its related mechanisms, since the identity of CM1 is still not clarified. In our previous study, we have already confirmed that soluble form of CM1 was produced by Raji. Therefore, we performed Q-TOF analysis after immunoprecipitation of concentrated Raji culture supernatant using anti-CM1 mAbs. Results: As a result, we found that CM1 is identical to enolase-1(ENO1), a glycolytic enzyme, and we confirmed that results by silencing ENO1 using siRNA. It was also confirmed through competition assay between anti-CM1 and anti-ENO1 mAbs. Finally, we investigated the possible role of CM1 in inflammatory response and cancer. The ligation of CM1 on Raji cells with anti-CM1 mAbs induces the extensive production of prostaglandin $E_2(PGE_2)$. In addition, the increased activity of matrix metalloproteinase (MMP)-2/9 was shown in NCI-N87, stomach cancer cell line by CM1 stimulation. Conclusion: CM1 is identical to ENO1 and it might be an important role in the regulation of inflammatory responses.

Relationship with Passage Time of Human Dental Pulp Stem Cells from Supernumerary Tooth by Classification (과잉치 분류에 따른 치수유래줄기세포 계대 배양 시간의 연관성)

  • Shin, Yeoseob;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.43 no.4
    • /
    • pp.419-426
    • /
    • 2016
  • For this research 20 supernumerary teeth impacted in the maxillary anterior have been extracted and pulp cells have been collected from them. From the collected pulp cells, total of 17 (10 males, 7 females) have been selected as subjects. From this research, the run-time of successive culture of the cell from tooth number pulp tissue was $2.91{\pm}0.29$ days. From the gathering of cells from the initial pulp tissue until gaining 80% confluency took $4.53{\pm}0.94$ which was the longest. The following successive cultures took $2.73{\pm}0.32$ days. Average runtime for female was $2.81{\pm}0.27$ days whereas male had average runtime of $2.98{\pm}0.29$ days. Average run-time for inversion was $2.94{\pm}0.30$ days and for normal location, $2.80{\pm}0.20$ days. Average runtime was $2.92{\pm}0.31$ days and other forms took $2.88{\pm}0.22$ days. In the future, follow up research would be needed to evaluate the efficiency of the cells collected from the initial passage and the latter passage as stem-cells and taking into consideration the less than 3 days'time for the subculture, it could be concluded that the research efficiency and fast cultivation would be sufficiently effective.

Transduced PEP-1-Grb7 Fusion Protein Suppressed LPS-induced COX-2 Expression

  • An, Jae-Jin;Kim, So-Young;Lee, Sun-Hwa;Kim, Dae-Won;Ryu, Hea-Jin;Yeo, Seung-Il;Jang, Sang-Ho;Kwon, Hyung-Joo;Kim, Tae-Yoon;Lee, Sang-Chul;Poo, Ha-Ryoung;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
    • /
    • v.40 no.2
    • /
    • pp.189-195
    • /
    • 2007
  • Although the incidence and severity of atopic dermatitis (AD) is steadily increasing at an alarming rate, its pathogenic mechanisms remain poorly understood yet. Recently, we found that the expression of Grb7 protein was markedly decreased in AD patients using proteomic analysis. In the present study, human Grb7 gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-Grb7 fusion protein. The expressed and purified PEP-1-Grb7 fusion proteins transduced efficiently into skin cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced PEP-1-Grb7 protein was stable for 48 h. In addition, transduced PEP-1-Grb7 fusion protein markedly increased cell viability in macrophage RAW 264.7 cells treated with LPS by inhibition of the COX-2 expression level. These results suggest that the PEP-1-Grb7 fusion protein can be used in protein therapy for inflammatory skin disorders, including AD.

Cloning of MT -hGH Gene-injected Rabbit Embryos by Nuclear Transplantation (사람성장호르몬 유전자주입 토끼수정란의 핵이식에 의한 복제)

  • Kang, T.Y.;Chae, Y.J.;Lee, H.;Park, C.S.;Lee, H.J.
    • Korean Journal of Animal Reproduction
    • /
    • v.22 no.4
    • /
    • pp.419-424
    • /
    • 1998
  • The present study was carried out to examine the efficiency of cloning of transgenic embryos by nuclear transfer(NT) using gene-injected rabbit embryos. The rabbit embryos at pronuclear stage were microinjected with methallothionein-human growth hormone(MT-hGH) gene and cultured to 8- and 16-cell in TCM-199 containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% $CO_2$incubator. The recipient oocytes were collected from the oviducts 14~16 h after hCG injection. The oocytes were enucleated and activated with 5$\mu$M ionomycin and 2mM 6-dimethylaminopurine. Blastomeres form gene-injected embryos were transferred into the enucleated oocytes by micromanipulation. The nuclear transplant oocytes were electrofused and co-cultured with rabbit oviductal cells. Following 120 h of culture, blastocysts were prepared for gene analysis by polymerase chain reaction(PCR). In previous experiment, the rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased while developing to blastocyst(25%)(Kang et al., 1998). The fusion rate of gene-injected blastomeres was significantly(P<0.05) lower than non-injected blastomeres(66% vs 80%). However, the NT embryos that were derived from gene-injected donor embryos did not differ from control embryos in development to the blastocyst stage(39% vs 31%). Of the 43 NT blastocysts developed from the gene-injected donor embryos, twelve(28%) were positive for the injected DNA. The results indicate that NT with gene-injected embryos can be successfully used for cloning and multiplication of transgenic embryos, furthermore applicable to improvement of transgenic animal production.

  • PDF

Detection of Astrovirus Infection from Hospitalized Young Children Feces by Reverse Transcription-Polymerase Chain Reaction (중증 위장관염 소아의 대변검체에서 역전사-중합효소 연쇄반응을 이용한 Astrovirus 검출)

  • Park, Hae-Kyung;Woo, So-Youn;Seoh, Ju-Young;Chong, Young-Hae;Seo, Jeong-Wan
    • The Journal of the Korean Society for Microbiology
    • /
    • v.34 no.5
    • /
    • pp.453-459
    • /
    • 1999
  • Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and co infection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows:1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 18 children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).

  • PDF

Gene Transfer into Pig and Goat Fetal Fibroblasts by Co-transfection of tPA Transgene and $Neo^r$ Gene

  • Kim, Bae-Chul;Han, Rong-Xun;Kim, Myung-Yoon;Shin, Young-Min;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
    • /
    • v.33 no.2
    • /
    • pp.107-111
    • /
    • 2009
  • The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.

Expression, Purification and Transduction of PEP-1-Botulinum Neurotoxin Type A (PEP-1-BoNT/A) into Skin

  • Kim, Dae-Won;Kim, So-Young;An, Jae-Jin;Lee, Sun-Hwa;Jang, Sang-Ho;Won, Moo-Ho;Kang, Tae-Cheon;Chung, Kwang-Hoe;Jung, Hyun-Ho;Cho, Sung-Woo;Choi, Jin-Hi;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
    • /
    • v.39 no.5
    • /
    • pp.642-647
    • /
    • 2006
  • Botulinum neurotoxin A (BoNT/A) has been used therapeutically to treat muscular hypercontractions and sudomotor hyperactivity and it has been reported that BoNT/A might have analgesic properties in headache. PEP-1 peptide is a known carrier peptide that delivers fulll-ength native proteins in vitro and in vivo. In this study, a BoNT/A gene were fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-BoNT/A fusion protein. The expressed and purified PEP-1-BoNT/A fusion proteins were efficiently transduced into cells in a time- and dose-dependent manner when added exogenously in a culture medium. In addition, immuno-histochemical analysis revealed that PEP-1-BoNT/A fusion protein efficiently penetrated into the epidermis as well as the dermis of the subcutaneous layer, when sprayed on mice skin. These results suggest that PEP-1-BoNT/A fusion protein provide an efficient strategy for therapeutic delivery in various human diseases related to this protein.

An experimental study on the cytotoxicity of orthodontic wires (교정용 호선의 세포독성에 관한 실험적 연구)

  • Lim, Yong-Kyu;Yang, Won-Sik
    • The korean journal of orthodontics
    • /
    • v.26 no.5 s.58
    • /
    • pp.591-599
    • /
    • 1996
  • This study was undertaken to investigate the cytotoxicity of orthodontic wires after doing various treatments to the wires. 018x025 inch Stainless steel(A) and Co-Cr(B) wires were used and each of them were divided into 4 groups. A-1 and B-1 groups were as received state, and A-2 and B-2 groups were heat treated. A-3 and B-3 groups were electropolished after heat treatment, and A-4 and B-4 groups were soldered with Ag-solder. Each group had 3 wires and these were sterilized with Ethylene Oxide gas. We used human gingival fibroblast cell culture and agar overlay technique to investigate the cytotoxicity of each group of wires. The cytotoxicity of wire was assessed using reaction index (zone index/lysis index). The findings of this study were as follows. 1. Both of the stainless steel wire and Co-Cr wire showed no cytotoxicity in as received state. 2. Heat treatment or electropolishing of the wires had no effect on the cytotoxicity of the wires 3. Soldered stainless steel wires showed a little wider zone of discoloration than soldered Co-Cr wires, but the zone index and cytotoxicity(reaction index) was not different. 4. Soldered wires showed moderate cytotoxicity in both of the wires.

  • PDF