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Effect of MEM Vitamins Supplementation of In vitro Maturation Medium and In vitro Culture Medium on the Development of Porcine Embryos

  • Kim, J.Y.;Lee, E.J.;Park, J.M.;Park, H.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.11
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    • pp.1541-1546
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    • 2011
  • This study was carried out to examine the influence of minimum essential medium (MEM) vitamins supplementation to in vitro maturation medium and in vitro culture medium on the development of porcine embryos. Porcine embryo development was investigated following cultivation in both in vitro maturation and culture medium with the supplementation of MEM vitamins (0, 0.1, 0.2 and 0.4%) using immature oocytes collected from the ovary of prepubertal gilts. Embryo development was observed and the total cell number in each blastocyst generated under the culture conditions was quantified following supplementation of the medium. The embryonic development rate of the blastocyst and hatched blastocyst was higher, but not significantly so, when 0.4% MEM vitamins were supplemented to the in vitro maturation medium of the porcine oocyte. Interestingly, the total number of cells in the blastocyst was significantly higher in the in vitro maturation MEM vitamins supplemented group compared to either the untreated group or the group which had MEM vitamins supplemented to both in vitro maturation and in vitro culture medium simultaneously (p<0.05). Therefore, the supplementation of 0.4% MEM vitamins to the in vitro mature medium has a beneficial effect on the embryonic development of in vitro produced blastocysts derived from the immature porcine oocytes.

The Effect of Hierarchy Culture on Clan Leadership and Organizational Commitment of Export-Driven SMEs

  • KIM, Hyuk Young
    • The Journal of Industrial Distribution & Business
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    • v.11 no.4
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    • pp.19-30
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    • 2020
  • Purpose: The purpose of this study examines the mediating effect of clan leadership in the relationship between hierarchy culture and organizational commitment. Most previous research focused on the relationship between organizational culture and organizational performance or organizational culture and job satisfaction. There are few empirical studies that focus on organizational commitment data because it is difficult to collect in many cases of export-driven small and medium sized enterprises. However, this research measures affective commitment, continuance commitment, and normative commitment differently than previous research, which is mostly focused on the hierarchy culture, clan leadership, and organizational commitment measurements. Research design, data, methodology: Conceptual research model is based on the studies of Cameron and Quinn (2011), and Gungor and Sahin (2018). The model is designed with three constructs such as hierarchy culture, organizational commitment, and clan leadership. The monitor culture and coordinator culture are as proxy for the hierarchy culture. The affective commitment, continuance commitment, and normative commitment are as proxy for the organizational commitment. And also the facilitator leadership and mentor leadership are as proxy for the clan leadership. Based on three hundred cases such as export-driven small and medium sized enterprises (SMEs), this study verify the hypothesis. Hypothesis was analyzed with the structural equation modeling. Results: In case of export-driven small and medium sized enterprises (SMEs), clan leadership acts as a mediator in the relationship between hierarchy culture and organizational commitment. In case of export-driven small and medium sized enterprises (SMEs) with high organizational commitment, clan leadership acts as a mediator in the relationship between hierarchy culture and organizational commitment. In case of export-driven small and medium sized enterprises (SMEs) with low organizational commitment, clan leadership did not act as a mediator in the relationship between hierarchy culture and organizational commitment. Conclusions: By controlling for the mediating effect of clan culture, this study have improved the academic contributions as well as policy and practical implications through empirical study of clan leadership that affect organizational commitment in the fields of hierarchy culture. In addition, this study means that the mediating effects on the variables of clan leadership were examined.

BIPHASIC CULTURE STRATEGY BASED ON HYPEROSMOTIC PRESSURE FOR IMPROVED HUMANIZED ANTIBODY PRODUCTION IN CHINESE HAMSTER OVARY CELL CULTURE

  • Kim, Min-Su;Kim, No-Su;Seong, Yun-Hui;Lee, Gyun-Min
    • 한국생물공학회:학술대회논문집
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    • pp.293-296
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    • 2002
  • Hyperosmotic pressure increased specific antibody productivity ($q_{Ab}$) of recombinant CHO cells (SH2-0.32) while it depressed cell growth. Thus, the use of hyperosmolar medium did not increase the maximum antibody concentration substantially. To overcome this drawback, the feasibility of biphasic culture strategy was investigated. In the biphasic culture, cells were first cultivated in the standard medium with physiological osmolality(294 mOsm/kg) for cell growth. When cells reached the late exponential phase of growth, the spent standard medium was replaced with the fresh hyperosmolar medium (522 mOsm/kg) for antibody production. The ($q_{Ab}$) in growth phase with the standard medium was 2.1 ${\mu}g/10^6cell/day$ while the ($q_{Ab}$) in antibody production phase with the hyperosmolar medium (522 mOsm/kg) was 11.1 ${\mu}g/10^6cell/day$. Northern blot analysis showed a positive relationship between the relative contenet of Ig mRNA and ($q_{Ab}$), indicating that transcriptional regulation was involved in the response of rCHO cells to hyperosmotic pressure. Due to the enhanced ($q_{Ab}$) and increased cell concentration in biphasic culture, the maximum antibody concentration obtained in biphasic culture with 522 mOsm/kg medium exchange was 161% higher than that obtained in batch culture with the standard medium. Taken together, simple biphasic culture strategy based on hyperosmotic culture for improved foreign protein production from rCHO cells is effective in improving antibody production of rCHO cells.

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The effect of medium change after pretreating microspores, medium addition, and volume of under solid medium in double layer culture on the production of embryos in isolated microspore culture of hot pepper (Capsicum annuum L.) (고추의 소포자 배양 시 전처리 후 배지의 교환, 배지의 첨가 및 2층배양 시 하층고체 배지의 양이 배의 생산에 미치는 영향)

  • Park, Eun-Joon;Lee, Jong-Suk;An, Dong-Joo;Kim, Moon-Za
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.494-504
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    • 2010
  • The effect of the addition of the fresh medium, volume of under solid medium in double layer culture as well as the medium change after pretreating microspores on the production of embryos in microspore culture of hot pepper (Capsicum annuum L.) has been studied. When cultured after heat pre-treatment, changing pretreatment media with fresh culture media proved to be more effective for embryo production rather than supplementing additional culture media. Heat-pretreating for 3 days turned out more effective for embryo production than pretreating for 1 or 2 days. In the case of anther pretreatment, the addition of fresh medium after culture was not effective for embryo production. In pretreating microspores, however, supplementing additional fresh culture media greatly improved embryo yield and quality. The best time point of media addition was 4 days after culture commenced, and the most effective number of times of media addition was one time addition. Moreover, the effective volume of added medium in double layer culture for embryo production was 1.5 ml. The addition of media more than 1.5 ml reduced both embryo yield and quality. Double layer medium was more effective for embryo development than liquid medium. When the volume of under solid medium increased ranging from 3 ml to 7 ml, more cotyledonary embryos were produced in either 5 ml or 7 ml compared to 3 ml, even though the total number of embryos were highest in 3 ml. These results can be used as an important data for establishing an efficient microspore culture system for producing high frequency of normal embryos in hot pepper.

Effects of Sucrose level and Nitrogen Source on Fresh Weight and Anthocyanin Production in Cell Suspension Culture of 'Sheridan' Grape (Vitis spp.)

  • Kim, Seung-Heui;Kim, Seon-Kyu
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.23-27
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    • 2002
  • To establish an in vitro mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of nitrogen source and sucrose on fresh weight and anthocyanin production in cell suspension culture of 'Sheridan' grape level were studied. When the medium was devoid of $NO_3^-$, cell fresh weight was either remained stable (1% sucrose) or slightly decreased with culture time (2,3, and 4% sucrose). When $NH_4^-$ was lacking, 3% sucrose was most favorable for cell growth. When $NH_4^-$ was supplied as N source, the anthocyanin content of 2% sucrose containing medium was maintained 2 times higher than other levels till day 8 in culture, then that of 3 and 4% sucrose which peaked at day 12 thereafter. The anthocyanin content was low than $NO_3^-$-free media. Total anthocyanin content in $NH_4^-$-free medium was just about a half of that of $NH_4^+$ medium. Anthocyanin production of 2% sucrose in $NH_4^+$ medium was maintained about 3-fold till day 8, then decreased thereafter. In $NH_4^+$ medium, pH decreased gradually with final pH of 3.5 to 4.0, while pH in $NH_4^+$-free medium increased with final pH of 6.5 to 7.5.

Effects of Trophoblastic Vesicle and Estradiol-$17\beta$ on the Development in Vitro of Rabbit Embryos (Trophoblastic Vesicle과 Estradiol-$17\beta$의 첨가가 가토배의 발달에 미치는 영향)

  • 오하식;박충생
    • Korean Journal of Animal Reproduction
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    • v.10 no.1
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    • pp.76-82
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    • 1986
  • This experiment was conducted to determine the effects of trophoblastic vesicles (TV) and estradiol-17$\beta$ on the development in vitro of rabbit embryos. Thirty matured female rabbits were treated with PMSG followed by HCG injection and mating. Embryos were recovered with D-PBS (Dulbecco's Phosphate Buffered Saline) after superovulation, and normally developed to two-to four-cell embryos were used in the subsequent in vitro culture. Basal medium was Medium-199 su, pp.emented with 1.5% bovine serum albumin. Embryo on Day 5 after mating (Day 0) was cut into two or three pieces to remove the embryonic disc. Each piece of tissue was cultured for 24 hours at 37$^{\circ}C$ in 0.5 mlMedium-199 in 5% CO2. During culture, peices of trophoblastic tissue changed into spherical vesicles which were used for co-culture. These spheres were called trophoblstic vesicles. Two-to four-cell embryos were cultured for 4 days in Medium-199 in the absence or presence of trophoblastic vesicle, and two-to four-cell embryos cultured with varing concentration (0, 0.1, 1, 10ng/ml) of estradiol-17$\beta$ for 4 dyas. Culture vessels used were watch glass for coculture with trophoblastic vesicles and micortube for estradiol-17$\beta$ infusion. Compared with the Medium-199 alone as basal culture medium, more blastocysts (46.7% vs 15.1%; P<0.01) and morulae (84.4% vs 56.6%; P<0.05) were developed in the co-culture with trophoblastic vesicles. Estradiol-17$\beta$ infused in culture medium was not effective for embryo development to blastocysts (78.3% in control, 50.0% in 0.1ng/ml, 61.5% in 1ng/ml and 64.4% in 10ng/ml) and also to morulae (91.3% in control, 84.2% in 0.1ng/ml, 92.3% in 1ng/ml and 91.1% in 10ng/ml). Compared with the watch glass culture mehotd, more (P<0.01) blastocysts were developed in microtube culture (78.3% vs 56.6%) and more (P<0.01) morulae in microtube culture (91.3% vs 56.6%).

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Invertase Production by Fed-batch Fermentations of Recombinant Saccharomyces cerevisiae

  • Koo, Ja-Hyup;Kim, Sang-Yong;Park, Yong-Cheol;Han, Nam-Soo;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.8 no.3
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    • pp.203-207
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    • 1998
  • Fed-batch fermentations with different feeding media were carried out in order to increase the productivity of invertase expression using a recombinant Saccharomyces cerevisiae containing plasmid pRB58. Two batch cultures showed the expression of the SUC2 gene at a low concentration of glucose, suggesting that glucose concentration could be used as a control variable in a fed-batch operation mode. In the fed-batch culture by feeding the basal medium, cell mass and specific invertase activity did not increase much as compared with the simple batch culture. A series of fed-batch cultures revealed that the sucrose-supplemented medium increased cell mass whereas the enriched medium did specific invertase activity. To capitalize on the synergism of the sucrose-supplemented medium and the enriched medium, the sucrose-supplemented enriched medium was used as a feeding medium. The fed-batch culture using this medium resulted in a 2.4-fold increase in cell mass and a 1.9-fold enhancement in specific invertase activity compared with those of the batch culture. The increase in cell mass and specific invertase activity led to a marked increase in total invertase activity, 250U/ml, which was 6.3 times higher than that of the batch culture.

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Serum Free Medium Development for Recombinant Erythropoietin Production using Novel Cell Line (QT35) (QT35 세포주에서 제조합 에리스로포이에틴 생산을 위한 무혈청 배지의 개발)

  • 주형민;김병기;김선영;김태한;김태용
    • KSBB Journal
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    • v.13 no.3
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    • pp.295-302
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    • 1998
  • Human Erythropoietin (EPO) gene is cloned in quail fibrosarcoma cell, QT35. Because molecular weight of EPO is similar to that of serum albumin, cell culture with serum containing medium makes purification of EPO very difficult. Using fractional factorial study, we have developed serum free medium for the recombinant QT35 cell lines, QT N4D4 and QT SY-IMP, which have cloned EPO with glutamine synthetase (GS) gene amplification system and with puromycin selective marker, respectively. Among the seven frequently used medium components, fibronectin, BSA, and EGF were the most important for EPO production. However, sufficient fibronectin supplement to the medium did not make any good attachment of QT35 to culture plate over 3 days. Therefore, to maximize EPO production, we attempted a medium-shift at confluence from serum containing medium to serum free medium(QT SFM6). Using the medium-shift protocol with QT SFM6, nearly the same productivity of EPO was achieved comparing with that without medium-shift. This result was true in both QT35 cell lines in three types of culture, i.e. T flask, microcarrier and roller bottle cultures.

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Development of Organ Culture Medium for Long Term Culture of Human Hair Follicle (인체 두피 모낭의 장기간 배양을 위한 기관 배양 배지의 개발)

  • Yoo, Bo-Young;Yoon, Hee-Hoon;Shin, Yeon-Ho;Seo, Young-Kwon;Lee, Doo-Hoon;Song, Kye-Yong;Hwang, Sung-Joo;Park, Jung-Keug
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.58-64
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    • 2006
  • We successfully isolated human anagen hair follicles from human scalp skin by microdissection and tried to culture them under various conditions. First we confirmed negative effect of serum on human hair follicle organ culture. As a next step serum-free medium compositions, Philpott medium, IMDM, and DHGM (Dongguk hair growth medium) were tried. Philpott medium is a general medium for hair organ culture based on Williams' E medium and DHGM is a special self-developed medium containing high amino acids and vitamins (B group) composition. As results, hair follicle in Philpott medium and IMDM showed anagen phase morphological structure, but rapid loss of hair elongation, low alkaline phosphatase expression, and very low expression of CK19. It is thought these hair follicles rapidly regressed from apoptosis. However, hair follicles in DHGM showed long term anagen phase morphological structure, continuous hair elongation, high alkaline phosphatase, and CK19 expression. These results demonstrate that high amino acids and vitamins (B group) composition are essential to in vitro long term human hair follicle organ culture and this culture medium will be useful in basic study of hair biology or application study to the development of alopecia treatment drugs.