• Korean Chemical Engineering Research
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• v.44 no.1
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• pp.73-80
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• 2006

This study compared cell expansion and colony forming ability in human cord blood stem cells cultured ex vivo with two kinds of cytokine combinations, two kinds of media, presence or absence of fetal bovine serum (FBS) and two or three dimensional (2D or 3D) culture environments. Purified $CD34^+$ cells were cultured in the IMDM (Iscove's Modified Dulbecco's Medium) and SFM (Serum Free Medium) containing a cytokine cocktail-I (coc-I) (EPO, GMCSF, SCF, and IL-3) or a cytokine cocktail-II (coc-II) (TPO, G-CSF, SCF, IL-6, and Flt3/Flk-2 ligand) with or without FBS. Generally, higher cellular and clonogenic expansion were observed in the coc-I cytokine condition, compared to coc-II cytokine condition. 3D (Methocult) and 2D (IMDM + coc-I + FBS) conditions gave the greatest cell ($2,258{\pm}456$ fold) and CFU (BFU-E: $652{\pm}19$, CFU-GM: $520{\pm}58$, CFU-GEMM: $339{\pm}100$ fold) expansions, respectively. In aspect of medium, IMDM was better than SFM, except for coc-II condition without FBS. In conclusion, 'IMDM + coc-I + FBS' and 'IMDM + coc-I' were the best CFU expansions on the occasion of all culture conditions. FBS and 2D conditions had affirmative effect on CFU expansion, generally. These data might provide a variety of notions about ex vivo expansion of hematopoietic stem cells.

• Journal of Embryo Transfer
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• v.20 no.1
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• pp.71-77
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• 2005

To investigate the influence of relaxin and insulin on the ovarian steroid secretion of porcine granulosa cells, we used porcine granulosa cells partially luteinized in a primary culture and examined the production of progesterone and $17{\beta}-estradiol$. Porcine granulosa cells were cultured in the presence of serum for 48 h after attachment and subsequently in the absence of serum fur 24 h. To confirm the dose dependency of relaxin or insulin, various concentrations (10, 100, 1000 ng/ml) of relaxin or insulin were added in the medium for the last 24 h, respectively. To investigate the combinational effect of relaxin and insulin, 100 ng/ml relaxin and/or 100 ng/ml insulin were added in the medium for the last 24 h in the presence or absence of luteinizing hormone (100 ng/ml). The medium was collected and used for radioimmunoassay to measure the production of progesterone and $17{\beta}-estradiol$. Relaxin or insulin increased the production of progesterone by dose dependency, respectively while they had no effect of the production of $17{\beta}-estradiol$. Relaxin (100 ng/ml) and/or insulin (100 ng/ml) significantly increased the production of progesterone in the presence of luteinizing hormone while they had no effect of the production of $17{\beta}-estradiol$. In conclusion, relaxin and/or insulin increased the progesterone secretion of porcine granulosa-lutein cells in vitro while had no effect on the production of $17{\beta}-estradiol$ and had no synergism on the effects. The effects of relaxin and/or insulin on the production of progesterone were augmented by the presence of luteinizing hormone.

• Proceedings of the Korea Water Resources Association Conference
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• 2006.05a
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• pp.552-556
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• 2006

항만건설에 있어서 항만의 내부시설 중에서 무엇보다도 가장 중요한 시설물은 선박을 안전하게 접안시켜 하역할 수 있는 안벽시설물이다. 안벽구조 형식의 결정은 항만의 이용목적 등에 따라서 달라지지만 항만의 건설입지조건 등에 의해서도 달라진다. 안벽구조형식 중에서 잔교 식 안벽은 무엇보다도 단기간에 건설이 용이하여 지금까지 각국에서 널리 사용되어 왔고 장래에도 이용도가 증가되리라 생각한다. 최근에는 해안선을 이용한 위락시설이 건설되면서 잔교 식 안벽구조물을 설치하여 보조시설물로 이용하는 경우가 많다. 과거에 설계되어 잔교를 설계할 경우는 일반적으로 항내의 정온이 잘 유지되는 경우에 대해서 설치하는 경우가 많기 때문에 파랑에 의한 반사율과 잔교 상부에 작용하는 양압력을 고려해야할 필요성이 거의 없었다. 그러나 최근에는 태풍이 내습할 경우 기존의 항내로 높은 파랑이 침입하는 경우가 발생하고 있어 항내에서도 잔교의 파괴로 인한 자연재해가 대형화되는 경우가 발생하고 있다. 또, 처음부터 안벽을 설계할 때에 대형화의 잔교 식 안벽구조물을 설치하는 경우도 있다. 이런 잔교 식 안벽 구조물을 잔교의 상부 판에 작용하는 양압력 분포와 잔교 전면의 반사율 등이 구조물의 유지관리 등에 미치는 영향이 매우 크기 때문에 반사율 검토와 양압력을 고려한 설계가 필요하다. 본연구의 대상은 일정 해역에 잔교 식 안벽을 설계하고자 할때 최적의 안벽 설계가 될 수 있도록 수리모형실험을 실시하여 구조물의 안전과 항내정온에 기초가 되는 자료를 도출하고자 하였다. 따라서 본 수리모형실험으로 인한 연구는 잔교 식 안벽에 대한 반사율과 상부에 작용하는 양압력, 잔교말뚝(pile)에 작용하는 수평압력을 검토하여 잔교 식 안벽 설계에 기초자료를 제공하고자 한다. 혈청을 이용한 동결보존을 대체할 수 있을 것으로 생각된다.지에 더해주면 세포의 증식이 개선될 것이다. 그래서 몇 가지 첨가물을 이용해 세포의 증식력에 변화가 나타나는지 알아보았다. 첨가물을 이용한 실험에서 IGF-I의 경우 장기간 배양에서 세포의 수를 안정적으로 유지하고 계대 횟수를 증가시키는 효과를 보였다. 이는 IGF-I이 어느정도 세포의 증식을 유지시켜주는 역할을 하기 때문인 것으로 생각된다. 무혈청 배지에서 비적응 CHO 세포의 계대 배양에 한계가 있는 것은 세포주기가 멈추기 때문인 것으로 생각된다. 세포주기가 멈추는 growth factor와 같이 세포의 증식을 지속적으로 유도할 수 있는 물질이 무혈청 배지에서는 부족하기 때문인 것으로 생각되고, IGF-I과 같은 첨가물을 통해 극복할 수 있는 문제라고 여겨진다.관점과 주거교육가치관 요소와의 관계를 알아본 결과, 전통적 관점은 주거교육가치관 요소 중 오직 주거관리적 요소와 관계가 있었으나 그 정도는 낮으며 실천적 관점과 구조적 관점은 주거가치관의 각 요소에 따라 약간 다르기는 했으나 주로 보통의 관계를 보였다.군 순으로 높게 관찰되었다. 이상의 결과를 종합할 때, 임상에서 니켈-티타늄 합금 와이어에 굴곡을 부여하기 위해 열처리하는 경우 초탄성 특성은 유지될 수 있으나, 부하-변위 곡선의 상방 증가가 나타나므로, 와이어에 의한 교정력이 증가될 수 있음에 유의하여야 한다. $day^{-1}$인 인공습지), scenario 2(면적 4.2ha인 저류지)가 각각 연평균 6.9%, 4.8%, 7.1%의 감소를 보였다. TN은 4.7%, 3.4%, 13.4%의 삭감율을 나타내었으며, TP는 5.6%, 3.9%, 7.3%의 삭감율을 나타내었다. 본 연구에서는

• 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.

• Journal of Life Science
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• v.33 no.1
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• pp.8-14
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• 2023

Peroxidasin (PXDN), a multidomain heme peroxidase containing extracellular matrix (ECM) motifs, as well as a catalytic domain, catalyzes the sulfilimine crosslink of collagen IV (Col IV) to reinforce Col IV scaffolds. We previously reported that PXDN is required for endothelial cell (EC) survival and growth signaling through sulfilimine crosslink-dependent matrix assembly. In this study, we examined whether peroxidase activity is required for PXDN function in ECs. First, we constructed a mutant PXDN by point mutation of two highly conserved amino acids, Q823 and D826, which are present in the active site of the peroxidase domain. After isolation of HEK293 clones highly expressing the mutant protein, conditioned medium (CM) was obtained after incubating the cells in serum-free medium for 24 hours and then analyzed by Western blot analysis under nonreducing conditions. The results revealed that the mutant PXDN formed a trimer and that it was cleaved by proprotein convertase-like wild-type (WT) PXDN. However, peroxidase activity was not detected in the CM containing the mutant PXDN, in contrast to that of WT PXDN. In addition, the sulfilimine crosslink ability of the mutant PXDN was lost. Moreover, the CM containing the mutant PXDN failed to promote the growth of PXDN-depleted ECs, unlike the CM containing WT PXDN. These results suggest that the peroxidase activity of PXDN affects EC growth by forming a sulfilimine crosslink.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.219-227
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• 2004

This study was conducted to develop a serum-free, defined medium of IVM of pig oocytes. The TCM-199 with supplemented with polyvinylalcohol(PVA), polyvinylpyrrollidone(PVP) and porcine follicular fluid(pFF) were used as basal medium. The effects of the these additives on the rates of maturity and development under in-vitro fertilization and in vitro culture were examined and subsequently considered on the possibilities be sustituted for the bovine serum albumin(BSA). Maturation rate of pig oocytes in IVM media containing PVA(82.4％), pFF(89.4％) and BSA(90.0％) were significantly higher(P<0.05) than that of PVP(78.6％). Cleavage rate after IVF of PVP(64％) was significantly lower(P<0.05) than these of PVA(73％), pFF(77％) and BSA(73％) supplements. in vitro development rates to morulae and blastocyst on PVP(54％) were also significantly lower(P<0.05) than these of the supplements of PVA(63％), pFF(69％) and BSA(65％). In comparison of maturation and fertilization rates of pig oocytes in each supplements, the maturity rates of PVA(82.4％), pFF(89.4％) and BSA(90.0％) were significantly lower(P<0.05) than that of PVP(72.4％) and while, the fertilization rates of pFF(87.1％) and BSA(89.1％) were significantly higher(P<0.05) than these of PVA(78.0％) and PVP(70.6％). It may be concluded that PVA and pFF can be substituted far BSA in medium for culturing pig oocytes; however, it may be considered that PVP were limited to for BSA in the in vitro culture of the embryos.

• Journal of Embryo Transfer
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• v.15 no.3
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• pp.231-236
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• 2000

This study was conducted to examine the effects of exogenous gonadotropins (PMSG+hCG) and an antioxidant (cysteine) on in vitro maturation of bovine follicular oocytes. Cumulus-oocyte complexes (COCs) aspirated from 2 to 5 mm ovarian follicles were cultured for 22 to 24 hours in a modified bovine embryo culture medium (mBECM) supplemented with 3 mg/mL bovine serum albumin, to which PMSG (10 IU/mL) + hCG (10 IU/mL) and/or cysteine (0.6 mM) were added. When examined the expansion of cumulus ce1ls at the end of maturation culture, greater (p<0.05) expansion was found after addition of PMSG+hCG (79 to 96%) to mBECM than after no addition (0%), regardless of the presence or absence of cysteine in the medium. The addition of cysteine did not stimulate cumulus expansion, but a high proportion (92%) of expansion was achieved when COCs were cultured after the addition of PMSG+hCG and cysteine to the medium. No difference in the proportion of oocytes underwent germinal vesicle breakdown (initiation of maturation) was found after the addition of PMSG+hCG and/or cysteine to mBECM. However, nuclear maturation (development to the metaphase-II stage) of oocytes was significantly stimulated by the combined addition of PMSG+hCG and cysteine, compared with no addition. In conclusion, both exogenous gonadotropins and an antioxidant are important for nuclear maturation of bovine immature oocytes and these factors have a cell-specific stimulatory action.

• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.389-394
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• 1994

8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.137-141
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• 2007

Antioxidants were well known to be essential supplements in the complex media and serve as a reservoir of oxygen. In this study, Hanwoo COCs (cumulus oocytes complexes) were matured and developed in L-cysteine-TCM199 and analyzed metaphase chromosome. Maturation rate of Hanwoo COCs were 73.4%, 94.6% in 0.1% PVA, 0.1 mM L-cysteine, respectively and showed significantly different between the treatments (p<0.05). Blastocyst formation were revealed 20.3%, 10.0% in 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA, respectively. There were no significant difference among treatment groups. Metaphase chromosome were showed 18.3%, 12.0% in 5% FBS-TCM199, 0.1 mM L-cysteine, respectively and analyzable chromosome were 6.1%, 4.0% and had no differences between the treated groups. In the case of in vitro developmental stages, metaphase chromosome were showed 18.3%, 12.0% in $4{\sim}16$ cells stage, 43.1%, 13.0% in morulae stage and 94.8%, 100.0% in blastocyst stage. These results suggested L-cysteine has beneficial role for in virto maturation and development in Hanwoo COCs.

• Food Science of Animal Resources
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• v.22 no.3
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• pp.274-280
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• 2002

This study was conducted to investigate inhibitory effects of K-casein, GMP and sialic acid addition on the infection of MA-104 cells by 597(Korean native cattle rotavirus) and JBR(Jeju island bovine rotavirus). MA-104 cells on incomplete Ml99 were infected with domestically separated 597 and ma activated by incubating at 37$\^{C}$ for 6 days, and analyzed for the titer of rotavirus. K-casein, GMP and sialic acid added MA-104 culture infected by activated S97 and nan were incubated for Is hours and stained by the AEC stainning method. The number of infected cells were counted on microscope. The titer of S97 and JBR was 2.5$\times$107 and 2.0$\times$106 PFU/ml, respectively. The inhibition level against cell infection by 597 was 97.4% far 2000UH of K-casein and 97.44% for 2000UM of GMP. The inhibition level against cell infection by JBR was 99.52% for 2000$\mu$M of $\kappa$-casein and 99.78% for 2000$\mu$M of GMP. The inhibition level against cell infection by 597 and JBR was 3.85 and 3.63% for 2000$\mu$M of sialic acid, respectively. The high inhibitory effects (over 97%) of K-casein and CMP against infection of U-1(14 cells with 597 and mR indicated great potentials for the use of K-casein and GMP in the treatment of calf or infant caused by rotavirus.