• KSBB Journal
• /
• 제16권1호
• /
• pp.30-35
• /
• 2001

유가식배양과 세라믹 막을 장착한 생물반응기에서 운전인 TCRC 조업을 통해서 Bacillus thuringiensis의 고농도 세포배양을 실시하였다. 유가식 배양에서 B. thuringiensis의 세포 성장은 선형적으로 증가하였고, 이것은 세포성장 모델리의 결과와 잘 맞았다. 낮은 세포 성장속에도 불구하고 유가식 배양동안에 포자형성은 관찰할 수 없었고, 이것은 연속배양의 결과와는 반대였다. 유가식 조업 후에 회분식 배양으로 바꾸면 300 g/L의 포도당 공급 농도를 사용했을 때 2.7$\times$$10^9$ CFU/mL 의 포자농도를 얻었다. 생물반응기내에 세라믹 막을 장착한 TCRC 조업에서 포도당 공급 농도의 영향을 결정하였다. 50 g/L의 포도당 농도를 사용했을 때 TCRC 조업에서 82.5 g-cell/L에 해당하는 최대 세포농도 1.8$\times$$10^{10}$ CFU/mL 를 얻었다. TCRC에서 세포성장은 선형적으로 증가하였고 포도당 농도는 제한되었는데 이것은 세포성장은 모델링의 결과와 잘 일치하였다. 1 g/L의 포도당을 공급한 경우 이외에는 TCRC 조업 동안에 포자형성을 관찰할 수 없었다. 50 g/L의 포도당을 공급한 경우 TCRC조업 후에 회분식 배양으로 전환시키면 1.2$\times$$10^{10}$ CFU/mL 의 포자농도를 얻었고, 이것은 연구된 여러 배양형태 중에 가장 높은 포자농도이다. 이 때 최적의 포도당 공급속도는 0.55 g glucose/h로 공급하였을 때였다.

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

• 대한한방부인과학회지
• /
• 제19권3호
• /
• pp.25-40
• /
• 2006

Purpose : This study was undertaken to evaluate the inhibitory effects of Arisaematis rhizoma on the cell proliferation in HeLa cells. Methods : The cultured cell after treatment in the different duration in 24, 48, 72 hours with solution of 1%. 5%, 10% Arisaematis rhizoma was quantified by trypan blue exclusin method. The control group was treated with 2% FBS in the different duration in 24, 48, 72 hours. We examined DNA of activated caspase by FACS analysis, caspase-3 activity, DNA fragmentation by DNA laddering, activity of HeLa Cells by the XTT assay, activity of MAP kinase by RT-PCR analysis. Results : After 72 hours culture, the growth activities of 1%, 5%, 10% Arisaematis rhizoma-treated Hela cell were significantly reduced with control group, respectively. After 24 hours culture, the ratio of cells showing caspase activity by FACS analysis were increased in 1%, 5%, 10% Arisaematis rhizoma-treated Hela cell. It were also increased in 48 hours culture of 10% and 72 hours culture of 5%, 10% Arisaematis rhizoma-treated Hela cell. In 24, 48 and 72 hours culture, DNA fragmentations of 5%, 10% Arisaematis rhizoma-treated Hela cell were obviously observed. These results meaned that Arisaematis rhizoma induces apoptosis of HeLa cells. It was supported by increased caspase-3 activity and decreased MAP kinase activity according to time periods and concentrations of Arisaematis rhizoma solution. Conclusion : The study shows that Arisaematis rhizoma has inhibitory effect on cell proliferation and induction capacity of apoptosis of human cevical carcinoma cell line, HeLa cells, in vitro. These results suggest that Arisaematis rhizoma should be useful for treatment of human cevical carcinoma.

• KSBB Journal
• /
• 제13권4호
• /
• pp.438-443
• /
• 1998

Effects of various environmental factors on cell size index(FCW/DCW) in Thalictrum rugosum. Lithospermum erythrorhizon and Taxus cuspidata plant cell suspension cultures were investigated. Time course change of cell size index were also observed. In batch cultures, FCW/DCW increased according to the decrease of sugar concentration. For short-term experiment within 24 hr, FCW/DCW value could be reduced significantly by increasing sugar concentration. When an osmoticum such as mannitol was added, FCW/DCW converged to a low value. Therefore, it was confirmed that osmolality of the medium was important in determining cell size or water content of the cells. Inorganic salts or treatment with organic solvent also exhibited some effect on the cell size index. However, pH and centrifugal force did not show any influences. On the other hand, it was found that the addition of Pluronic F-68 reduced FCW/DCW. By combining these results effectively, it may be possible to increase the cell concentration in high density culture to a higher extent.

• Journal of Nutrition and Health
• /
• 제53권6호
• /
• pp.563-569
• /
• 2020

Purpose: The vascular smooth muscle cells (VSMCs) in mature animals have implicated to play a major role in the progression of cardiovascular diseases such as atherosclerosis. This study aimed at optimizing the protocol in culturing primary VSMCs (pVSMCs) from rat thoracic aorta and investigating the effect of cellular zinc (Zn) deficiency on cell proliferation of the isolated pVSMCs. Methods: The thoracic aorta from 7-month-old Sprague Dawley rats was isolated, minced and digested by the enzymatic process of collagenase I and elastase, and then inoculated with the culture Dulbecco Modified Eagle Medium (DMEM) at 37℃ in an incubator. The primary cell culture morphology was observed using phase-contrast microscopy and cellular Zn was depleted using Chelex-100 resin (extracellular zinc depletion only) or 3 µM N,N,N',N'-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN) (extracellular and intracellular zinc depletion). Western blot analysis was used for the detection of SM22α and calponin as smooth muscle cell marker proteins and von Willebrand factor as endothelial cell marker protein to detect the culture purity. Cell proliferation by Zn depletion (1 day) was measured by MTT assay. Results: A primary culture protocol for pVSMCs from rat thoracic aorta was developed and optimized. Isolated cultures exhibited hill and valley morphology as the major characteristics of pVSMCs and expressed the smooth muscle cell protein markers, SM22α and calponin, while the endothelial marker von Willebrand factor was hardly detected. Zn deprivation for 1 day culture decreased rat primary vascular smooth muscle cell proliferation and this pattern was more prominent under severe Zn depletion (3 µM TPEN), while less prominent under mild Zn depletion (Chelexing). Conclusion: Our results suggest that cellular Zn deprivation decreased pVSMC proliferation and this may be involved in phenotypic modulation of pVSMC in the aorta.

• 대한치과보철학회지
• /
• 제42권3호
• /
• pp.261-266
• /
• 2004

Statement of problem. The effects of surface roughness have not or insufficiently been analyzed on earlier events such as cell adhesion though cell behavior most germane to implant performance is cell adhesion. Purpose. The purpose of this study was to evaluate cell adhesion of osteoblast-like cells (MG63) onto three types of titanium disks with varying roughness using the Elisa assay. Materials and methods. Representative disks from each group (SLA, HA, machined) were subjected to surface analysis and surface roughness was measured by the optical interferometer (Accura 2000, Intekplus Co., Seoul, Korea). Following this, MG63 cells were cultured on the titanium disks and released. Cell adhesion measurements using the Elisa assay were performed specifically at three points: after 24, 48, and 72 hours of culture. Results. Among the 3 types of surface analyzed, the SLA surface was the roughest with a Ra value of $1.114{\mu}m$ followed by HA coated surface and machined surface, consecutively. The optical density values for the SLA surface group was significantly higher than that of the machined and HA coated surface groups following 24 and 48 hours of culture. The cell culture on HA coated surface showed significantly higher values compared to the machined surface following 24, 48 and 72 hours of culture. Conclusion. The results suggest that surface treatment of titanium surfaces enhanced cell adhesion of human osteoblast-like cells (MG63).

• KSBB Journal
• /
• 제31권1호
• /
• pp.66-72
• /
• 2016

Chinese hamster ovary (CHO) cells have been widely used for production of various recombinant proteins such as cytokines and monoclonal antibodies. The cell aggregation and cell death in CHO cell culture directly affect cell viability, and productivity and quality of products. In this study, we investigated preventing effects of storage-protein 2 (SP2) derived from silkworm hemolymph on cell aggregation and cell death in CHO cell culture producing albuminerythropoietin (Alb-EPO). The viable cell density in the culture supplemented with 2 mg/mL SP2 was 1.71-fold higher than that in control culture. Increased titer of Alb-EPO was also found in the culture with SP2. Morphology of CHO cells in SP2 supplemented cultures did not differ from that of control. In addition, the cell aggregation rate of the SP2 cultures was reduced 20% compared to the control. Finally, we confirmed that the apoptosis was strongly suppressed by addition of SP2 in the cultures. These results clearly demonstrate that SP2 can be served as an effective supplement for enhancing titer of Alb-EPO via reducing cell aggregation and cell death.

• KSBB Journal
• /
• 제8권5호
• /
• pp.478-482
• /
• 1993

LSM을 이용하여 KA112 균주의 고농도 배양을 시도하였다. Separator로는 hollow fiber를 사용하였고 reactor로는 Celligen을 이용하였다. Wroking volume 1리터로 10일간 배양하여 최고 세포농도가 회분식 배양에 비하여 10배 이상 증가한 $2.1\times10^7$ cells/ml이었고, 항체의 농도는 4.5배 정도 높았다. 최고 feed rate에서 항체생산속도는 회분식 배양보 다 9배 높았으며 배양 중 glucose농도가 Ig/e 이상일 때 specific productivity가 증가하였고, 1 g/6 이하얼 때 세포성장은 영향을 받지 않으냐 spe­c cific prodictivity는 감소하였다.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제7권2호
• /
• pp.117-120
• /
• 2002

The effect of cell density on cell growth was investigated in a suspension batch culture of hybridoma cells. The specific growth rate was found to increase with increasing initial cell density and then to decrease with further increases in initial cell density. In order to quantitatively describe the dependence of specific growth rate on cell density, a kinetic model is proposed, which satisfactorily represents the experimental data.

• 한국발생생물학회지:발생과생식
• /
• 제19권3호
• /
• pp.119-126
• /
• 2015

The suitable feeder cell layer is important for culture of embryonic stem (ES) cells. In this study, we investigated the effect of two kinds of the feeder cell, MEF cells and STO cells, layer to mouse ES (mES) cell culture for maintenance of stemness. We compare the colony formations, alkaline phosphatase (AP) activities, expression of pluripotency marker genes and proteins of D3 cell colonies cultured on MEF feeder cell layer (D3/MEF) or STO cell layers (D3/STO) compared to feeder free condition (D3/-) as a control group. Although there were no differences to colony formations and AP activities, interestingly, the transcripts level of pluripotency marker genes, Pou5f1 and Nanog were highly expressed in D3/MEF (79 and 93) than D3/STO (61and 77) or D3/- (65 and 81). Also, pluripotency marker proteins, NANOG and SOX-2, were more synthesized in D3/MEF ($72.8{\pm}7.69$ and $81.2{\pm}3.56$) than D3/STO ($32.0{\pm}4.30$ and $56.0{\pm}4.90$) or D3/- ($55.0{\pm}4.64$ and $62.0{\pm}6.20$). These results suggest that MEF feeder cell layer is more suitable to mES cell culture.