• Microbiology and Biotechnology Letters
• /
• v.28 no.2
• /
• pp.117-123
• /
• 2000

The object of this work is to improve the maintenance of bioluminescence from stored Photobacterium phosphoreum in a view of developing continuous monitoring system for pollutants. The long-term experiments were performed to determine the effect of storage temperature and immobilization on the maintenance of bioluminescence and viability of P. phosphoreum. A naturally luminescent bacterium, P. phosphoreum was starved in 2.5% Nael solution at $20^{\circ}C$, $4^{\circ}C$, -$20^{\circ}C$ and -$70^{\circ}C$ for 30 days. In vivo luminescence was measured by luminometry, and total cell concentrations and concentrations of culturable and viable cells were determined by acridine orange staining, dilution plate counting, and direct viable counting, respectively. The bioluminescence emission from cells stored at 4De was maintained up to 10 days while those with starved cells at other temperature ranges decreased to background level within 3 days. In terms of viability of cells, concentrations of cells stored at $20^{\circ}C$ were rapidly decreased as a result of cell lysis, leading to a drop in culturable and viable counts while cells stored at $4^{\circ}C$ was shown viable but nonculturable state during starvation. With immobilized cells on strontium alginate, the bioluminescence showed higher maintenance than free cells and decreased with count number of nonculturable cells.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.21 no.6
• /
• pp.527-535
• /
• 2020

The purpose of this study was to establish a simple vitrification protocols to preserve animal cell lines derived from tissues of livestock that could be recultured. Bovine oviduct epithelial cells (BOEC) were used for the vitrification process using a 0.25 ml straw to increase cryopreservation efficiency. BOEC was cultured from the oviduct of 3.5-day estrus state, and the commercially available polyampholyte StemCell Keep^TM was used as a cryoprotective agent. Using different concentrations, the viability rates of BOEC in 5, 10, 25, 50, 75, and 100% in freezing media were investigated. Survivability was determined using a differential staining technique using a trypan blue test and a CYTO-13/PI staining protocol. The viability rates of BOEC in the trypan blue test were 5.6±11.8, 12.5±7.2, 53.0±2.7, 85.1±6.9, 79.8±0.6, and 60.7±6.7% with a respective concentration of StemCell Keep^TM. The viability rates in CYTO-13/PI staining were 4.6±2.5, 30.8±12.1, 58.4±2.5, 85.5±1.2, 79.8±0.6, and 71.2±1.2%, respectively. These results indicate that BOEC could be preserved with StemCell Keep^TM without toxicity in a 0.25-ml straw. The optimal concentration of vitrification solution with StemCell Keep^TM was determined to be 50% and can be considered as a proper preservation method for cryobanking.

• Journal of Life Science
• /
• v.26 no.2
• /
• pp.226-233
• /
• 2016

Vascular smooth muscle cell (VSMC) apoptosis has been identified in various vascular diseases, including atherosclerosis and restenosis after angioplasty, and has been known to precipitate atherosclerotic plaque instability and rupture. Oxysterols are known as inducers of apoptosis in VSMC, and 7-ketocholesterol (7KC) is the major nonenzymically formed oxysterol in atherosclerotic lesions. The precise mechanism underlying VSMC apoptosis is still poorly understood. In this study, we investigated whether 7KC causes apoptosis, and characterized its apoptotic mechanisms in primary cultured rat aortic VSMC. Cell viability was assessed by MTT assay and trypan blue assay. Apoptosis was assessed by flow cytometry, immunofluorescence, immunoprecipitation, and Western blot analyses. 7KC markedly decreased the VSMC viability in a time- and concentration-dependent manner, and increased the production of 4-hydroxynonenal (HNE), a major end-product of lipid peroxidation, which also decreased the VSMC viability. Pretreatment with 2,4-dinitrophenylhydrazine, a well-known reagent of lipid peroxidation-derived aldehydes, significantly restored the 7KC-decreased viability of VSMC. Furthermore, HNE, as well as 7KC, reduced the level of total Akt, a major mediator of cell survival. The 7KC-decreased level of total Akt was significantly restored by pretreatments with 2,4-dinitrophenylhydrazine and N-acetylcysteine. Lactacystin, a proteasome inhibitor, protected VSMC against apoptosis and Akt degradation, but did not inhibit HNE production. In the immunoprecipitation assay, 7KC increased HNE-modified Akt. From the results, it seems that, in atherosclerotic lesions, 7KC induces HNE production in VSMC, and this HNE binds to Akt, proceeding to proteasomal degradation of Akt, through which mechanism the atherosclerotic plaque instability may be facilitated.

• Journal of Plant Biology
• /
• v.36 no.4
• /
• pp.399-406
• /
• 1993

유채(Brassica napus L. cv. Topas) 소포자 배양에 의한 기내 배발생 과정을 관찰하고 발생과정에 따른 총단백질의 변화 양상을 이차원 전기영동을 통하여 분석하였다. 배양에 적합한 소포자의 발달 단계는 4,6-diamidino-2-phenylindole (DAPI) 형광염색으로 핵상관찰을 통하여 결정하였다. 최초의 소포자 생존율은 63.9%였으며 이러한 생존율의 차이로 배양초기에 소포자의 이형성이 나타났다. 배 발달 경로에 따른 최초의 세포분열은 화분벽 안에서 균등분열로 시작되어 배병의 발달이 선행된 후 배 발생이 일어났으며, 구형, 심장형, 어뢰형으로의 배 발달은 여러 조직의 분화와 더불어 빠르게 진행되었다. 소포자배 발달과정을 치상단계의 소포자, 배양 3일째의 초기 분열세포, 구형 및 심장형배, 어뢰형배, 성숙한 자엽단계의 배 등의 5단계로 나누어 각각 2차원 전기영동을 수행한 결과, 최초 소포자 단계에서 나타났던 23개의 단백질들은 배 달달 경로로 진행됨에 따라 사라지고, 배양 3일째에는 8개의 단백질이 특이적 또는 지속적으로 발현되었다. 배의 발달과 더불어 20∼50 kD 사이에서 총 42개의 단백질이 급격히 나타나거나 또는 후기 배로 진행하면서 점차적으로 발현되었다.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.19 no.1
• /
• pp.155-165
• /
• 2006

이 연구는 MCF-7 인간 유방암 세포주에 대한 귀출파징탕(歸朮破癥湯) 추출물의 증식억제효과 세포독성효과 세포사 유발효과를 확인하기 위하여 이루어졌다. MCF-7 인간 유방암 세포주는 Dulbecco's modified Eagle's medium/F12 (DMEM/F12)에 10% fetal bovine serum(FBS;Gibco) 와 항생제를 가하여 만든 배지를 이용하여 배양하였고, MCPF-7 세포를 96- well plate에 접종한 후 다양한 농도(0~2000g/ml)의 귀출파징탕(歸朮破癥湯)이 든 배지로 처리하고 다양한 시간(48, 96, 192)동안 배양하여 현미경으로 관찰하고 각각 MTS assay kit를 이용하여 세포생존율을 측정하였다. 세포독성은 Sulforhodamine B assay 방법을 이용해 측정하였고 세포사 과정에서 MCF-7세포에서의 caspase 활성화를 측정하기 위해 Western blotting을 수행하여 poly ADP ribose polymerase(PARP)의 절단을 확인하였다. 실험결과 귀출파징탕(歸朮破癥湯) 추출물에 의한 세포성장 및 독성효과는 시간 및 농도에 비례하는 것으로 나타났고 세포고사과정에서 작용하는 caspase의 전 기질인 PARP 절단량이 귀출파징탕(歸朮破癥湯) 처리 농도와 시간에 비례하여 증가하였다. 이것은 caspase-3가 MCF-7 세포의 성장을 억제하는데 중요한 역할을 수행함을 의미한다. 따라서 귀출파징탕(歸朮破癥湯)은 다양한 기전에 의해서 유방암 세포에 대한 억제효과를 가진다는 것을 인식할 수 있다.

• Applied Biological Chemistry
• /
• v.50 no.1
• /
• pp.63-67
• /
• 2007

The neuroprotective effect of methanolic extracts from Dendrobium nobile Lindl. (DME) against $H_{2}O_{2}$-induced neurotoxicity in PC12 cells was investigated. The treatment of PC12 cells with various DME concentrations under $H_{2}O_{2}$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay, an LDH release assays, and a morphological assay. Interestingly, we also detected reduction of apoptotic bodies and inhibition of caspase-3 activity by DME in $H_{2}O_{2}$-indeced PC12 cells. These data show that the neuroprotective effect of DME against PC12 cells might be related to the suppression of caspase-3 activation. Therefore, these results suggest that DME could be a new potential candidate as chemotherapeutic agents against neuronal diseases.

• The Journal of the Convergence on Culture Technology
• /
• v.8 no.3
• /
• pp.291-296
• /
• 2022

This study was conducted on curcumin which had increased bioavailability as a potential AChE inhibitor for the treatment of neurodegenerative diseases. The purpose of this study is to confirm the in vitro neuroprotective effect on Aricumin (turmeric extract). To confirm the neuroprotective effect, AChE inhibition for Aricumin was evaluated, and cell viability was analyzed for HT-22cell, and oxidative stress (glutamate, H₂O₂)-induced HT-22 cytotoxicity was evaluated. As a result of the change in the AChE inhibition rate of Aricumin (Turmeric extract), it was confirmed that Aricumin at a concentration of 39.06㎍/ml or higher inhibited AChE activity by about 20% and more. And it was confirmed that the cytotoxicity of HT-22 cells induced by oxidative stress (Gluamate 5 mM and H₂O₂ 500 µM) was significantly inhibited from 0.01 to 0.1 mg/ml concentration (p<005). These results suggest that Aricumin (turmeric extract) have potential neuroprotective effects.

• Korean Journal of Clinical Laboratory Science
• /
• v.56 no.1
• /
• pp.73-84
• /
• 2024

A pulsed electromagnetic field (PEMF) enhances the efficacy of several anticancer drugs. Doxorubicin (DOX) is an anticancer agent used to treat various malignancies, including breast cancer. This study examined whether a PEMF increases the anticancer effect of DOX on MCF-7 human breast cancer cells and elucidated the underlying mechanisms affected by PEMF stimulation in DOX-treated MCF-7 human breast cancer cells. A cotreatment with DOX and a PEMF potentiated the reduction in MCF-7 cell viability compared to the treatment with DOX alone. The PEMF elevated DOX-induced G1 arrest by affecting cyclin-dependent kinase 2 phosphorylation and the expression of G1 arrest-related molecules, including p53, p21, cyclin E2, and polo like kinase 1. In addition, PEMF increased the DOX-induced upregulation of proapoptotic proteins, such as Fas and Bcl-2-associated X, and the downregulation of antiapoptotic proteins, including myeloid leukemia 1 and survivin. PEMF promoted the DOX-induced activation of caspases-8, -9, and -7 and poly (adenosine diphosphate-ribose) polymerase cleavage in MCF-7 cells. In conclusion, PEMF enhances the anticancer activity in DOX-treated MCF-7 breast cancer cells by increasing G1 cell cycle arrest and caspase-dependent apoptosis. These findings highlight the potential use of a PEMF as an adjuvant treatment for DOX-based chemotherapy against breast cancer.

• Journal of the Korean Applied Science and Technology
• /
• v.34 no.2
• /
• pp.296-304
• /
• 2017

The purpose of this study was to investigate the applicability of the Psidium guajava leaf extract as a whitening functional cosmetic material. We measured DPPH radical scavenging activity, intracellular ROS, cytotoxicity in B16F10 melanoma cells and cytoprotective effect on ultraviolet A, in vitro tyrosinase inhibitory effect and melanin biosynthesis inhibitory effect. The antioxidative effect was confirmed through high DPPH radical scavenging activity and intracellular ROS activity inhibition measurement of the Psidium guajava leaf extract. The survival rate of B16F10 melanoma cells was more than 98% at all concentrations, and the cytoprotective effect from ultraviolet ray A was found to increase in a concentration-dependent manner. In addition, in vitro tyrosinase activity inhibitory effect of 10% and melanin biosynthesis inhibitory effect of 20% were observed. Through less toxicity for B16F10 melanoma cell, high antioxidant activity, inhibition of tyrosinase activity and melanin biosynthesis inhibitory effect, we confirmed the possibility of developing the Psidium guajava leaf extract as a whitening functional cosmetic material with a safe and excellent whitening effect.

• Journal of Life Science
• /
• v.22 no.5
• /
• pp.621-626
• /
• 2012

$Drosophila$ $melanogaster$ has been used as a useful model to study development and disease. In this study, we established the primary culture method of $Drosophila$ in the intestine to understand how intestinal stem cells (ISCs) mediate tissue repair during infection and disease. To obtain intestinal cells, we separated intestines from adult flies and isolated single cells by enzymatic treatment. The survival of cultured cells was measured using MTS-analysis. The maximum growth rate of the cells was observed on the 9th day after seeding. In addition, the presence of ISCs and enteroendocrine cells was confirmed by delta and prospero staining. Accordingly, we supposed that $Drosophila$ $melanogaster$ gut cells established in this study are probably useful in studies about intestinal stem cell regulation and various diseases occurring in the intestine.