• ALGAE
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• 제32권1호
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• pp.67-74
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• 2017

The heterotrophic dinoflagellate Oxyrrhis marina is known to produce high levels of docosahexaenoic acid (DHA) when fed on diverse algal prey. However, large-scale culturing of algal prey species is not easy and requires a large amount of budget, and thus more easily cultivable and low-cost prey is required. Dried yeast was selected as a strong candidate for an alternative prey in our preliminary tests. Thus, we explored the fatty acid composition and DHA production of O. marina fed on dried yeast and compared these results to those of O. marina fed on two algal prey species: the phototrophic dinoflagellate Amphidinium carterae and chlorophyte Chlorella sp. powder. O. marina fed on dried yeast, which does not contain DHA, produced the same high level of DHA as those fed on DHA-containing A. carterae. This indicates that O. marina is likely to produce DHA by itself regardless of prey items. Furthermore, the DHA content (and portion of total fatty acid methyl esters) of O. marina satiated with dried yeast, 52.40 pg per cell(and 25.9%), was considerably greater than that of O. marina fed on A. carterae (26.91 pg per cell; 15.7%) or powder of Chlorella sp. powder (21.24 pg per cell; 16.7%). The cost of dried yeast (approximately 10 US dollars for 1 kg dried yeast) was much lower than that of obtaining the algal prey (approximately 160 US dollars for 1 kg A. carterae). Therefore, compared to conventional algal prey, dried yeast is a more easily obtainable and lower-cost prey for use in the production of DHA by O. marina.

• 한국축산식품학회지
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• 제42권1호
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• pp.175-185
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• 2022

This study investigated the amino acid and nucleotide-related compound composition and taste characteristics of cultured muscle tissue (CMT) obtained by culturing satellite cells isolated from chicken and cattle and compared them to those of traditional meat (TM). The content of all amino acids except valine and tyrosine was significantly different between CMT and TM (p<0.05). The amount of glutamic acid was not significantly different between CMT and TM in cattle, but the glutamic acid in chicken CMT was lower than that of TM (p<0.05). Among the nucleotide-related compounds, only the content of inosine-5'-monophosphate (IMP) was significant, and the amount of IMP in CMT derived from chicken and cattle was significantly lower than that of TM (p<0.05). There were significant differences in the taste characteristics assessed by an electronic tongue system, and the umami, bitterness, and sourness values of CMT were significantly lower than those of TM from both chicken and cattle (p<0.05). The results of the present study suggest that it is necessary to develop a satellite cell culture method that could increase the umami and bitterness intensity of CMT and adjust the composition of the growth medium to produce cultured meat with a taste similar to that of TM.

• Journal of Ginseng Research
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• 제46권1호
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• pp.167-174
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• 2022

Background: 20(S)-protopanaxadiol (20(S)-PPD), one of the main active metabolites of ginseng, performs a broad spectrum of anti-tumor effects. Our aims are to search out new strategies to enhance anti-tumor effects of natural products, including 20(S)-PPD. In recent years, fasting has been shown to be multi-functional on tumor progression. Here, the effects of fasting combined with 20(S)-PPD on hepatocellular carcinoma growth, apoptosis, migration, invasion and cell cycle were explored. Methods: CCK-8 assay, trypan blue dye exclusion test, imagings photographed by HoloMonitorTM M4, transwell assay and flow cytometry assay were performed for functional analyses on cell proliferation, morphology, migration, invasion, apoptosis, necrosis and cell cycle. The expressions of genes on protein levels were tested by western blot. Tumor-bearing mice were used to evaluate the effects of intermittent fasting combined with 20(S)-PPD. Results: We firstly confirmed that fasting-mimicking increased the anti-proliferation effect of 20(S)-PPD in human HepG2 cells in vitro. In fasting-mimicking culturing medium, the apoptosis and necrosis induced by 20(S)-PPD increased and more cells were arrested at G0-G1 phase. Meanwhile, invasion and migration of cells were decreased by down-regulating the expressions of matrix metalloproteinase (MMP)-2 and MMP-9 in fasting-mimicking medium. Furthermore, the in vivo study confirmed that intermittent fasting enhanced the tumor growth inhibition of 20(S)-PPD in H22 tumor-bearing mice without obvious side effects. Conclusion: Fasting significantly sensitized HCC cells to 20(S)-PPD in vivo and in vitro. These data indicated that dietary restriction can be one of the potential strategies of chinese medicine or its active metabolites against hepatocellular carcinoma.

• 생명과학회지
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• 제33권9호
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• pp.724-729
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• 2023

본의 연구 결과는 무기질 구리가 전사, 번역 및 번역 후의 여러 단계에서 아펠린 생합성에 억제 효과를 발휘한다는 것을 명확하게 보여준다. 그러나 바토쿠프로인디설폰산의 구리 킬레이터로 처리하면 구리의 억제 영향이 효과적으로 역전되어 포유류 골격근 세포에서 아펠린 생합성의 구리 의존적 특성이 확인되었다. 이러한 결과는 구리가 마이오카인 아펠린의 생합성 조절에 중요한 역할을 한다는 최초의 보고이며, 구리 관련한 근육감소증의 예방 및 치료의 전략 개발에 유용한 실마리를 제공할 수 있다.

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.39-46
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• 2000

Objective: A number of studies to improve in vitro culture conditions have been tried over past ten years by using co-culture system with helper somatic cells. However, the mechanism of coculture is poorly understood. This study was designed to understand the mechanism for the mode of actual action of co-culture using co-culture system of ICR strain's 1-cell embryos with human oviduct epithelial cells by examining the effect of conditioned medium and contactless coculture using a cell culture insert on the embryo development and by measuring the level of superoxide anion from conditioned medium after co-culture. Methods: ICR strain's zygote embryos were cultured in medium alone (control), coculture, conditioned medium, or contactless coculture system for 6 days. Conditioned media (CM) were prepared as following 5 groups. All CM were collected after culturing oviduct cells for 2 days. CM-1 was stored at $-20^{\circ}C$ until use, and CM-2 was prepared just before use as a culture medium. CM-3 was cocultured with embryos and retrieved just before use. CM-4 and CM-5 were derives from the microfilteration of CM-2 and CM-3, respectively, using Microcon-10 (10 kDa molecular weight cut-off). The percentage of the embryos developed to hatched blastocyst stage and the level of superoxide anion in supernatant from medium alone culture (control), coculture, and contactless coculture were measured. Results: The rates of embryo development to the hatched blastocyst stage were significantly higher in coculture (43%) than in control (0%) (p<0.05). The CM-1 group had no embryo development since 2-cell embryonic stage, whereas the CM-2, CM-3, CM-4 and CM-5 groups had the improved development to 4 or 8 cell embryo stage, but the similar rate of development to hatched blastocyst compared to control. The effect of coculture on embryo development was disappeared in the contactless coculture group. The level of superoxide anion was significantly reduced in coculture group compared to control. Conclusion: It is concluded that the present coculture system overcomes the 2-cell block in vitro and improves the embryo development. This beneficial effect may be due to the direct cell-cell contact between embryo and helper cells or the removal of deleterious components from medium rather than the embryotrophic factors.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권2호
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• pp.100-107
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• 2004

Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage ($31.0%{\pm}8.09%$ in P4 group to $65.5%{\pm}24.08%$ in P2 group), compared with that obtained in the abscence of Ara-C ($17.6%{\pm}6.03%$) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to $56.22%{\pm}0.67%$ and GFAP positive cells to $66.46%{\pm}1.83%$ in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권1호
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• pp.42-51
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• 2006

Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage (31.0%${\pm}$8.09% in P4 group to 65.5%${\pm}$24.08% in P2 group), compared with that obtained in the abscence of Ara-C (17.6%${\pm}$6.03%) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to 56.22%${\pm}$0.67% and GFAP positive cells to 66.46%${\pm}$1.83% in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

• 한국버섯학회지
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• 제2권1호
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• pp.15-20
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• 2004

국내에서 많이 재배되는 버섯 중의 하나인 느타리 P. ostreatus는 먹물버섯이나 치마버섯과는 달리 실험실 수준에서의 자실체 발생이 힘든 것으로 알려져 있다. 본 연구에서는 실험실에서 느타리버섯을 샤레를 이용하여 전 발생과정을 유도하기 위한 방법을 검토하였다. 샤레상의 배양조건은 프라스틱 샤레 ($60{\times}15mm$)의 감자배지에서 균사를 접종한 후 빛이 없는 상태에서 배양한 뒤에, 균사의 환기상태, 균사표면의 상처, 빛 그리고 저온충격 등의 여러 환경요인들이 원기 및 자실체 형성에 미치는 영향을 검토하였다. 느타리의 최초 자실체 형성은 접종 이후 3주 내에 얻을 수도 있었으며, 균사접종 이후 10주 동안에 자실체의 형성은 균주에 따라서 76%에서 97%의 높은 빈도로 유도될 수 있었다. 위와 같이 샤레상에서 자실체를 형성할 수 있었으며, 정상적인 자실체의 성장을 위해서 빛은 필수적이며, 환기도 필요하였다. 또한 균사의 상처 처리가 원기, 자실체 및 포자의 형성에 미치는 영향이 균주에 따라서 크게 차이가 났으며, 같은 균주라 하더라도 발생단계별 그 반응의 차이도 크다는 사실을 확인하였다. 이들 자실체에서 수집된 담자포자는 발아가 가능하였다. 샤레상의 완전한 자실체 형성 방법은 느타리버섯의 생육주기를 단축시킬 수 있을 뿐만 아니라, 다양한 재배조건의 시료 획득과 한개의 배양 용기 내에서 버섯 발생의 전 단계를 관찰하고 분석하는데 유용할 수 있다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권7호
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• pp.980-987
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• 2008

The objective of this study was to investigate the effects of cell status of BOEC on development of bovine IVM/IVF embryos and gene expression in BOEC before or after culturing of embryos. The developmental rates beyond morula stage in the BOEC co-culture group was significantly higher than in the control group (p<0.05). In particular, blastocyst production in the BOEC co-culture group (28.3%) was dramatically increased compared with the control group (7.2%). In the in vitro development of bovine IVM/IVF embryos according to cell status, the developmental rates beyond morula stage in the primary culture cell (PCC) co-culture group were the highest of all experimental groups. Expression of genes related to growth (TGF-${\beta}$ EGF and IGFBP), apoptosis (Bax, Caspase-3 and p53) and antioxidation (CuZnSOD, MnSOD, Catalase and GPx) in different status cells of BOEC for embryo culture was detected by RT-PCR. While EGF gene was detected in isolated fresh cells (IFC) and PCC, TGF-${\beta}$ and IGFBP were found in IFC or PCC after use in the embryo culture, respectively. Caspase-3 and Bax genes were detected in all experimental groups regardless of whether the BOEC was used or not used in the embryo culture. However, p53 gene was found in IFC of both conditions for embryo culture and in frozen/thawed culture cells (FPCC) after use in the embryo culture. Although antioxidant genes examined were detected in all experimental groups before using for the embryo culture, these genes were not detected after use. This study indicated that the BOEC co-culture system used for in vitro culture of bovine IVF embryos can increase the developmental rates, and cell generations and status of BOEC might affect the in vitro development of bovine embryos. The BOEC monolayer used in the embryo culture did not express the growth factors (TGF-${\beta}$ and EGF) and enzymatic antioxidant genes, thereby improving embryo development in vitro.

• 한국안광학회지
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• 제12권3호
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• pp.19-25
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• 2007

현재 시중에서 유통되고 있는 소프트 콘택트렌즈 관리용액인 다목적용액(Multi Purpose Solution, MPS)이 결막세포주(Clone 1-5C-4 cell line)에 미치는 증식저해정도와 가토안의 각막상피 및 내피조직에 미치는 손상정도를 비교 관찰하고자 시행하였다. MPS는 $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea)를 사용하였다. 세포증식 저해율은 결막세포주를 배양 한 후 MTT assay로 검정하였고, 형태학적으로는 광학현미경으로 Hematoxylin & Eosin staining 표본을 제작하여 관찰하였다. In vivo 실험은 백색 가토 9마리(18안)를 3군으로 분류하여 실험군인 좌안(9안)에는 각 MPS제품을, 대조군인 우안(9안)에는 보존제가 포함되지 않은 멸균생리식염수를 접안하였다. 일정기간 접안 후, 가토안의 각막표변을 Rose bengal로 염색하여 관찰하였고 각막상피 및 내피조직의 변화는 주사전자현미경(Scanning Electron Microscopy, SEM)으로 비교 관찰하였다.