• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.712-720
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• 2007

To investigate clonal variations of recombinant Chinese hamster ovary(rCHO) clones in response to culture pH and temperature, serum-free suspension cultures of two antibody-producing CHO clones(clones A and B), which were isolated from the same parental clone by the limiting dilution method, were performed in a bioreactor at pH values in the range of 6.8-7.6, and two different temperatures, $33^{\circ}C\;and\;37^{\circ}C$. In regard to cell growth, clone A and clone B displayed similar responses to temperature, although their degree of response differed. In contrast, clones A and B displayed different responses to temperature in regard to antibody production. In the case of clone A, no significant increase in maximum antibody concentration was achieved by lowering the culture temperature. The maximum antibody concentration obtained at $33^{\circ}C$(pH 7.4) and $37^{\circ}C$(pH 7.0) were $82.0{\pm}2.6$ and $73.2{\pm}4.1{\mu}g/ml$, respectively. On the other hand, in the case of clone B, an approximately 2.5-fold increase in maximum antibody concentration was achieved by lowering the culture temperature. The enhanced maximum antibody concentration of clone B at $33^{\circ}C$($132.6{\pm}14.9{\mu}g/ml$ at pH 7.2) was due to not only enhanced specific antibody productivity but also to prolonged culture longevity. At $33^{\circ}C$, the culture longevity of clone A also improved, but not as much as that of clone B. Taken together, CHO clones derived from the same parental clone displayed quite different responses to culture temperature and pH with regards antibody production, suggesting that environmental parameters such as temperature and pH should be optimized for each CHO clone.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.203-209
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• 2009

The present study was performed to identify changes of plasminogen activator (PA) and plasminogen activator inhibitor (PAI) in porcine oviduct epithelial cells (POECs) during the estrous cycle. POECs obtained from ovary in pre-ovulatory (Pre-Ov), early to mid-luteal stage (Early-mid L) and post-ovulatory stage (Post-Ov). For the examine of PA activity, $1{\times}10^5$ fresh cells of POECs were cultured in DMEM/Ham F-12 containing 10% FBS and 0.2% amphotericin under humidified atmosphere of 5% $CO_2$ in air and $38^{\circ}C$. The urokinase-type PA (uPA) was observed at 7 days of POECs culture. PA activity was measured with culture prolonged of 0, 3, 6, 12 and 24 h after culture of 7 days. The PA activity were high significantly (p<0.05) at 12 h of culture, but PA activity were decreased with culture periods increased. The PA activity in POECs of Post-Ov stage were higher significantly (p<0.05) than that of Early-mid L and Pre-Ov stage. When PAI-1 and PAI-2 were added during the POECs culture, the PA were observed significant low activity (p<0.05). The PA activity and protein expression were decreased by PA inhibitor. This results suggest that PAI-1 and PAI-2 have a suppressive action on change of PA activity during the estrous cycle of pigs. Specifically, this study using PA inhibitor was effect the PA activity and PAI expression in oviduct epithelial cells in pigs.

• Toxicological Research
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• v.22 no.3
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• pp.301-306
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• 2006

Microcystin-LR(MC-LR), a cyanobacterial toxin produced by Microcystis aeruginosa, causes severe hepatotoxicity. Here we investigated the morphologic changes of rat hepatocyte spheroid induced by exposure of MC-LR($10^{-6}M$) in vitro. In addition, to determine the effects of such toxin in the process of hepatocyte spheroid formation, primarily isolated hepatocytes were incubated with MC-LR and the process of spheroid formation was observed. In both hepatocyte spheroid and suspension culture systems, the morphologic changes caused by MC-LR were noticible at 5 min post exposure and were characterized by the loss of microvilli, cytoplasmic vacuolation, the accumulation of lipid droplets, and blob formation. Especially, the size and numbers of blob on the cell surface were increased as the incubation time prolonged and the appearance of electron dense bodies were observed in the cytoplasm of hepatocyte at 20 min post exposure. Furthermore, bile canaliculi-like structures in the hepatocyte spheroids were slightly widened and the process of spheroids formation was inhibited in the isolated hepatocytes incubated with MC-LR. These results indicate that morphologic changes in. the hepatocyte membrane and organelles seem to be typical events in showing the MC-LR induced hepatotoxic effects and the spheroid culture method might be a useful experimental tool to evaluate hepatoxicity since it reflects the in vivo status of hepatocytes.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1208-1212
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• 2007

Production of extracellular epothilone B, one of the potent anticancer agents, by free and immobilized Sorangium cellulosum was studied using the repeated batch culture process. The concentration of alginate used in immobilization was directly related to the mass transfer rate of nutrients, mechanical stability, and the epothilone B production yield. With the optimized 3% (w/v) calcium alginate carrier, a prolonged repeated batch culture was investigated for the 5 repeated batches for 24 days. The maximum productivity of epothilone B obtained from the alginate-immobilized cells was 5.03 mg/l/day, which is 3 times higher than that of free cells (1.68 mg/l/day).

• Journal of Plant Biotechnology
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• v.6 no.4
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• pp.213-219
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• 2004

Zygotic embryos were excised from immature and mature seeds of the Himalayan yew, Taxus wallichiana. The embryos germinated precociously when kept in darkness for 5 weeks and developed into full seedlings within 10-12 weeks. The highest rate of embryo germination ($81\%$) was obtained in modified Lloyd & McCown' s woody plant medium containing macro and micronutrients at half strength supplemented with $1\%$ activated charcoal, which supported both the best embryonic growth ($43\%$) and seedling development ($32\%$). However, the supplementation of basal media with kinetin, thidiazuron, 6-benzyl aminopurine or $GA_3$ had no effect on the germination of the embryos. The embryos derived from immature seeds germinated but the frequency of embryonic growth was better in mature seeds. Stratification of seeds effected precocious germination of embryos. Seeds kept at $4^{\circ}C$ for 1 week germinated earlier and at a higher frequency irrespective of the stage of seed maturity, while the germination rate declined with prolonged cold treatment for 1 month at that same temperature. Analysis of taxanes in germinating seedlings revealed that root tissues contained high levels of taxol, 10-deacetyl-baccatin ill and baccatin ill as compared to shoots. Thus embryo culture technique appears to overcome the lengthy dormancy requirement of T. wallichiana seeds.

• Animal cells and systems
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• v.2 no.2
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• pp.229-232
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• 1998

Lysophosphatidylcholine (LPC) has been reported to be responsible for the sustained activation of protein kinase C (PKC). As chondroqenesis is known to be regulated by PKC, this study was performed to investigate the effects of LPC on chondrogenesis of chick limb bud mesenchymes in vitro. LPC treatment of mesenchymes during micromass culture significantly enhanced chondrogenic differentiation. The most effective time of LPC on the stimulation of chondrogenesis was the first day of micromass culture. Analysis of LPC effects on the expression of PKC isoforms revealed that LPC treatment increased expression of PKCa, among the multiple PKC isoforms, in the membrane fraction on day one of culture. The stimulatory effect of LPC on chondrogenesis was abolished if PKCa was down regulated by the prolonged treatment of cells with phorbol ester. The results suqqest that LPC promotes chondrogenesis through the activation of PKCa at the early stage of chondrogenic differentiation.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.277.2-277.2
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• 2013

Petri dishes and glass slides have been widely used as general substrates for in vitro mammalian cell cultures due to their culture viability, optical transparency, experimental convenience, and relatively low cost. Despite the aforementioned benefit, however, the flat two-dimensional substrates exhibit limited capability in terms of realistically mimicking cellular polarization, intercellular interaction, and differentiation in the non-physiological culture environment. Here, we report a protocol of culturing embryonic rat hippocampal neurons on the electro-spun polymeric network and the results from examination of neuronal cell behavior and network formation on this culture platform. A combinatorial method of laser-scanning confocal fluorescence microscopy and live-cell imaging technique was employed to track axonal outgrowth and synaptic connectivity of the neuronal cells deposited on this model culture environment. The present microfiber-based scaffold supports the prolonged viability of three-dimensionally-formed neuronal networks and their microscopic geometric parameters (i.e., microfiber diameter) strongly influence the axonal outgrowth and synaptic connection pattern. These results implies that electro-spun fiber scaffolds with fine control over surface chemistry and nano/microscopic geometry may be used as an economic and general platform for three-dimensional mammalian culture systems, particularly, neuronal lineage and other network forming cell lines.

• International Journal of Advanced Culture Technology
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• v.9 no.3
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• pp.100-105
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• 2021

This study examines the effectiveness of the study through a case of PBL(problem-based-learning) class conducted in a balanced culture course called at 00- University in the second semester of 2020. The effects of learning are as follows: First, PBL(problem-based-learning) has sufficient active interaction between the teacher and the learner. In the face of prolonged non-face-to-face learning, the PBL teaching method has sufficient interaction between the professors-learner and the learner. Second, PBL learning can actively utilize various problems that fit the characteristics of the subject and actively utilize the process of role sharing and collaboration. By presenting various problem situations suitable for the subject, students will be able to share roles individually or as a team, and fully experience the process of collaboration and discussion in the process of investigating the data. Third, critical perceptions of problem situations can be extended. In modern times, a variety of problem situations arise and critical perceptions of them must be fully learned. In a mass production and mass consumption society, students should develop the ability to blindly recognize and distinguish between real and fake information in a flood of information. The limitations identified in this class case are, first, the nature of the subject, "Understanding Culture and Philosophy," which makes it possible to discuss the global cultural phenomenon, but it should be discussed in terms of philosophy. Second, it is not easy to work as a team on non-face-to-face online. Nevertheless, PBL is a very effective method of learning in which active interactions and learning activities take place between professors and students, whether face-to-face or face-to-face online learning.

• Journal of the Ergonomics Society of Korea
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• v.33 no.2
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• pp.155-165
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• 2014

Objective: The aim of this study is to develop an office chair enabling to keep working at reclined sitting posture. Background: Sedentary workers are supposed to change the posture frequently during long hours of sitting. A reclined sitting position has been recommended to reduce disc pressure. But slumped sitting posture caused by the buttock sliding forward without any adjustment of back reclining is commonly observed. The worker seems to have tendency to change the sitting posture maintaining working condition. We assumed the reason to be their hands movement away from the working space when tilting backward. Method: Slide mechanism allowing seat to move forward was designed to maintain the hand position in working space during reclining. A prototype was manufactured and tilting motion was analyzed using motion capture system. Four experiment chairs were tested including the manufactured prototype chair and three other commercial chairs. Results: A backward movements of the hand position were 13.0mm, 101.7mm, 156.1mm and 139.3mm at the prototype chair, compared to chair B, chair C and chair D, respectively. And the movement was remarkably small at the prototype chair. Conclusion: The developed seat sliding chair allows back tilting maintaining hand position at working space. We expect the user tilting back more often than normal tilting chair during seated work. But further investigation is required to figure out the effectiveness of the developed chair using prolonged working hours. Application: The developed office chair directly affects commercialization.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.369-373
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• 2003

In spite of potential benefits of anther culture, low productivity of plant regeneration in some genotypes; e.g. tonsil and indica rice, is one of the major obstacles for practical use of anther culture. This study was conducted to improve cold shock method and carbohydrate source for increasing the efficiency of anther culture in rice. The most common carbon source, sucrose was replaced to maltose, which has two molecules of glucose. Maltose increased callus induction 1.4-to 1.8-fold higher in japonica rice, 3.2-to 11.6-fold in tongil types and 2.7-fold in indica rice IR50. Callus induction was increased from 0.2％ to 12.5％ in maltose medium compared to the medium supplemented with sucrose plus glucose in indica rice "Tetep". A simple procedure of vacuum packaging of panicles during cold shock treatment prolonged not only anther viability more than 15 days but also increased callus induction more than 2-fold compared to open-air storage (conventional method). Combining of above two methods, callus induction was increased 28 to 56％ in japonica, 13 to 33％ in tonsil type and 12 to 31％ in indica rice. Plant regeneration was increased 14 to 35％ in japonica, 10 to 20％ in tonsil and 4 to 15％ in indica rice, respectively.