• Title/Summary/Keyword: two-stage culture

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Betaine Production by Two Stage Culture and Elicitor in the Cell Cultures of Lycium chinense Mill (구기자 현탁 세포배양으로부터 이단계 배양과 Elicitor에 의한 Betaine 생산)

  • 김병원;노광수
    • KSBB Journal
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    • v.13 no.5
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    • pp.569-576
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    • 1998
  • The effects of carbohydrates, hormones and elicitors on both cell growth and betaine production were investigated in the cell cultures of Lycium chinense Mill. The maximum effect of glucose and sucrose was observed in cells cultured in the presence of 3% and 7% for cell growth and betaine production, respectively. The effect of hormones on cell growth and betaine production was prominent in the presence of 10 ${\mu}$M 2, 4-D, 10 ${\mu}$M NAA and 2.5 ${\mu}$M IAA, whereas cell growth and betaine production were excellent at 2.5 ${\mu}$M BA and 10 ${\mu}$M BA, respectively. Abiotic elicitors such as KCI, MnCl2 and NaCl exhibited an inhibitory role on cell growth in all treatment groups. Betaine production was increased according to increase of concentration of abiotic elicitors. methanol-soluble and insoluble components as biotic elicitor remarkably inhibited cell growth from 2 mg and 6 mg, respectively. Betaine production was increased maximally at 2 mg of biotic elicitors. When growth medium was switched to production medium at two stage culture, it resulted that cell fresh weight and dry weight decreased but betaine content increased about 2.2-fold.

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Optimization of Xylitol Production by Candida tropicalis in Two-stage Fed-batch Culture (Candida tropicalis의 2단계 유가식 배양에 의한 Xylitol 생산의 최적화)

  • 유연우;조영일;서진호
    • KSBB Journal
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    • v.17 no.1
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    • pp.93-99
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    • 2002
  • Two-stage fed-batch culture of Candide tropicalis that was designated primarily to cultivate the cell in the glucose medium (1st stage) and then produced the xylitol from xylose medium (2nd stage) was developed to improve a xylitol yield and productivity. In the growth stage, glucose was automatically supplied to the fermentor by pH-stat mode when the pH was up 5.7, When a feeding medium was added in order to reach the glucose and yeast extract concentrations up to 100 and 40 g/L, respectively, a high cell concentration and a relatively low ethanol concentration were obtained in 18.5 h culture. In the production stage, initial xylose concentration of 150 g/L was the most favorable for obtaining the final xylitol concentration and productivity. The addition of mineral salts was also enhanced a xylitol production. But the aeration rate was not significantly affected a xylitol production. When the addition of 16 g yeast extract and 232.5 g xylose powder at the production stage was used, xylitol yield and productivity were significantly increased. With these conditions, xylitol concentration, yield and productivity of 108.9 g/L, 74%) and 3.3 g/L·h, respectively, were obtained in a final volume of 1.58 L. The further addition of 16 g yeast extract and 232.5 g xylose powder increased the working volume partly (1.67 L) and resulted in a relatively high xylitol concentration, yield and productivity of 193 g/L, 70% and 3.6 g/L·h, respectively.

Enhancement of preimplantation mouse embryo development with optimized in vitro culture dish via stabilization of medium osmolarity

  • Hyejin Yoon;Jongwoo Lee;Inyoung Kang;Kyoo Wan Choi;Jaewang Lee;Jin Hyun Jun
    • Clinical and Experimental Reproductive Medicine
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    • v.50 no.4
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    • pp.244-252
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    • 2023
  • Objective: We evaluated the efficacy of the newly developed optimized in vitro culture (OIVC) dish for cultivating preimplantation mouse embryos. This dish minimizes the need for mineral oil and incorporates microwells, providing a stable culture environment and enabling independent monitoring of individual embryos. Methods: Mouse pronuclear (PN) zygotes and two-cell-stage embryos were collected at 18 and 46 hours after human chorionic gonadotropin injection, respectively. These were cultured for 120 hours using potassium simplex optimized medium (KSOM) to reach the blastocyst stage. The embryos were randomly allocated into three groups, each cultured in one of three dishes: a 60-mm culture dish, a microdrop dish, and an OIVC dish that we developed. Results: The OIVC dish effectively maintained the osmolarity of the KSOM culture medium over a 5-day period using only 2 mL of mineral oil. This contrasts with the significant osmolarity increase observed in the 60-mm culture dish. Additionally, the OIVC dish exhibited higher blastulation rates from two-cell embryos (100%) relative to the other dish types. Moreover, blastocysts derived from both PN zygotes and two-cell embryos in the OIVC dish group demonstrated significantly elevated mean cell numbers. Conclusion: Use of the OIVC dish markedly increased the number of cells in blastocysts derived from the in vitro culture of preimplantation mouse embryos. The capacity of this dish to maintain medium osmolarity with minimal mineral oil usage represents a breakthrough that may advance embryo culture techniques for various mammals, including human in vitro fertilization and embryo transfer programs.

A Novel Simple Method to Purify Recombinant Soluble Human Complement Receptor Type 1 (sCR 1) from CHO Cell Culture

  • Wang, Pi-Chao;Hisamune Kato;Takehiro Inoue;Masatoshi Matsumura;Noriyuki Ishii;Yoshinobu Murakami;Tsukasa Seya
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.67-75
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    • 2002
  • The human complement receptor type 1 (CR 1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently; it is reported that CRl was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CRl, called sCRl, is a recombinant CRl by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCRl losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity, furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.

Berberine Production by Cell Suspension Cultures of Cork Tree (Phellodendron amurense Rupr)

  • Choi, Myung-Suk;Shin, Dong-Ill;Park, Young-Goo
    • Korean Journal of Pharmacognosy
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    • v.27 no.1
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    • pp.32-36
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    • 1996
  • Various culture conditions for cell growth and berberine production in cork tree (Phellodendron amurense Rupr.) were investigated. Callus was induced from cambium tissue of cork tree, and cultured on LS liquid medium supplemented with 0.5 mg/1 2,4-D, 0.1mg/1 BA, and 3% sucrose. Several factors enhancing berberine production and cell growth in cork tree cell cultures were found. Some of them enhanced both cell growth and berberine production, but others resulted in a decoupling of cell growth and berberine production with significant in the specific levels. High level of nitrate (80mM), high level of phosphate (8.98mM), and sucrose (7%), 1.0mg/l IAA were effective in berberine production, whereas low level of nitrate (40mM), and phosphate (2.25mM), and high level of sucrose (7%) in the medium were effective in cell growth. Two stage culture(first stage for cell growth, and second stage for berberine production) increased berberine production almost twice (5.06mg/g dry weight) as much as single stage cultures in berberine production.

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Mouse Embryo Culture used in Quality Control of Water for Human in Vitro Fertilization : The One-cell Stage Versus the Two-cell Stage Model (수질에 대한 1-세포기 및 2-세포기 생쥐배아를 이용한 생물학적 정도관리에 관한 연구)

  • Lee, Ye-Kyung;Chung, Hye-Won;Kim, Hyung-Mee;Oh, Seung-Eun;Son, Young-Soo;Yu, Han-Ki;Woo, Bock-Hee
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.1
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    • pp.9-17
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    • 1993
  • This study was carried out investigate the effect of water quality and the kind of media on the in vitro development of 1-cell and stage mouse embryos. $F_1$ hybrid mice were superovulated and timely mated. 1-cell stage and 2-cell stage mouse embryos were recruited and taken into Ham's F-10 or m-KRB media which was made of two of two kinds of water having different quality, highly purified water and tap water. 2-cell stage embryos grew up well in vitro to blastocyst or hatching blastocyst regardless of the composition of culture media, but 1-cell stage mouse embryo didn't develop well to blastocyst or hatching blastocyst in simple media like m-KRB. These results meant in vitro devleopment of 1-cell stage mouse embryo neded complex media like Ham's F-10 which contained abundant protein components. In case of quality control for water, in vitro fertilization program. observation of in vitro development of 2-cell mouse embryos up to blastocyst or hatching blastocyst media such as m-KRB would be efficatious in detecting the difference of water quality.

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Effect of Osmolarity of Culture Medium on the Preimplantation Development of Porcine NT and IVF Embryos

  • Hwang, In-Sun;Park, Mi-Rung;Moon, Hyo-Jin;Shim, Joo-Hyun;Kim, Dong-Hoon;Yang, Byoung-Chul;Ko, Yeoung-Gyu;Yang, Boh-Suk;Cheong, Hee-Tae;Im, GI-Sun
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.91-96
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    • 2007
  • In vitro development of porcine embryo is affected by culture condition. One possible factor is osmolarity of culture medium. 1his study examined whether high osmolarity of culture medium at the early culture stage improves development of preimplantation porcine in vitro fertilization (IVF) and nuclear transfer (NT) embryos. NT and IVF embryos were divided into three groups and the basic medium was PZM-3 ($250{\sim}270$ mOsmol, control group). The control group of embryos was cultured in PZM-3 for whole culture period. Other two groups of embryos were cultured in a modified PZM-3 with 0.05 M sorbitol or 0.05 M sucrose ($300{\sim}320$ mOsmol, sorbitol or sucrose group) for the first 2 days, and then cultured in PZM-3 for further culture. NT embryos cultured in sucrose group showed a significantly higher developmental rate to the blastocyst stage with a decreased apoptosis rate compared to the sorbitol (p<0.05). For IVF, sucrose group showed a significantly increased the blastocyst formation rate with a decreased apoptosis rate compared to the control (p<0.05). This study represents that the high osmolarity in the early embryo culture stage can enhance the in vitro development of porcine NT and IVF embryos to the blastocyst stage with reduced apoptosis of cells.

Large-Scale Production of Cronobacter sakazakii Bacteriophage Φ CS01 in Bioreactors via a Two-Stage Self-Cycling Process

  • Lee, Jin-Sun;Kim, Gyeong-Hwuii;Kim, Jaegon;Lim, Tae-Hyun;Yoon, Yong Won;Yoon, Sung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.31 no.10
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    • pp.1430-1437
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    • 2021
  • Cronobacter sakazakii is an opportunistic pathogenic bacterium found in powdered infant formula and is fatal to neonates. Antibiotic resistance has emerged owing to overuse of antibiotics. Therefore, demand for high-yield bacteriophages as an alternative to antibiotics has increased. Accordingly, we developed a modified mass-production method for bacteriophages by introducing a two-stage self-cycling (TSSC) process, which yielded high-concentration bacteriophage solutions by replenishing the nutritional medium at the beginning of each process, without additional challenge. pH of the culture medium was monitored in real-time during C. sakazakii growth and bacteriophage CS01 propagation, and the changes in various parameters were assessed. The pH of the culture medium dropped to 5.8 when the host bacteria reached the early log phase (OD540 = 0.3). After challenge, it decreased to 4.65 and then recovered to 4.94; therefore, we set the optimum pH to challenge the phage at 5.8 and that to harvest the phage at 4.94. We then compared phage production during the TSSC process in jar-type bioreactors and the batch culture process in shaker flasks. In the same volume of LB medium, the concentration of the phage titer solution obtained with the TSSC process was 24 times higher than that obtained with the batch culture process. Moreover, we stably obtained high concentrations of bacteriophage solutions for three cycles with the TSSC process. Overall, this modified TSSC process could simplify large-scale production of bacteriophage CS01 and reduce the unit cost of phage titer solution. These results could contribute to curing infants infected with antibiotic-resistant C. sakazakii.

The Globalization and Corporate Culture of Nestle and Samsung Electronics in Vietnam (다국적기업의 현지화를 위한 경영문화 연구 - 베트남 진출 네슬레와 삼성전자의 사례 비교 -)

  • Lee, Sing-Young;Kim, Hyoun-Chul
    • International Commerce and Information Review
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    • v.9 no.4
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    • pp.375-393
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    • 2007
  • The purpose of this study is to define the different stage of global management cultures for localization from the case study of two respective multinational corporations, Nestle and Samsung Electronics invested in Vietnam, and then to find out on how Korean multinational corporations can reduce the gap of global management culture compared to global multinational corporations. In fact, Samsung has been pretty much being on a right track from the global marketing management perspective, however still stand at the beginning stage in terms of leading the global corporate culture. In comparison to Nestle with over 130 years of worldwide business experience, Samsung Electronics having only a decade global experience as a Korean multinational corporation has been found to have an obvious gap in terms of globalization which is mainly caused by global corporate culture. Even though it doesn’t seem realistic for Korean multinational corporations to immediately catch up the gap of global corporate culture against global multinational corporations with long years of global experiences and history, Korean multinational corporations need to track the foot print of every steps of globalization particularly focused on the local management culture of global multinational corporations, so that they will be able to shorten the timing to develop the true meaning of global corporate culture being evolved from the Korean management style.

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Xylitol의 생산성 향상을 위한 Two-stage Fed-batch 배양조건의 최적화

  • Jo, Yeong-Il;Seo, Jin-Ho;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.356-359
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    • 2000
  • A two-stage fed-batch fermentation was carried out to increase xylitol productivity by Candida tropicalis. The first stage for cell growth was performed in the pH-stat and continuous fed-batch modes. The higher cell growth and lower ethanol production obtained in the fed-batch mode where the growth medium was fed when pH of culture broth increased over 5.7. And also the effect of oxygen transfer on xylitol production was investigated by changing agitation speed under 0.5 vvm of aeration. The maximum xylitol productivity and yield were obtained at 500 rpm of agitation.

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