• Animal Bioscience
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• 제36권11호
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• pp.1757-1768
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• 2023

Objective: The number of bovine mammary epithelial cells (BMECs) is closely associated with the quantity of milk production in dairy cows; however, the optimal levels and the combined effects of hormones and essential amino acids (EAAs) on cell proliferation are not completely understood. Thus, the purpose of this study was to determine the optimal combination of individual hormones and EAAs for cell proliferation and related signaling pathways in BMECs. Methods: Immortalized BMECs (MAC-T) were treated with six hormones (insulin, cortisol, progesterone, estrone, 17β-estradiol, and epidermal growth factor) and ten EAAs (arginine, histidine, leucine, isoleucine, threonine, tryptophan, lysine, methionine, phenylalanine, and valine) for 24 h. Results: Cells were cultured in a medium containing 10% fetal bovine serum (FBS) as FBS supplemented at a concentration of 10% to 50% showed a comparable increase in cell proliferation rate. The optimized combination of four hormones (insulin, cortisol, progesterone, and 17β-estradiol) and 20% of a mixture of ten EAAs led to the highest cell proliferation rate, which led to a significant increase in cell cycle progression at the S and G2/M phases, in the protein levels of proliferating cell nuclear antigen and cyclin B1, cell nucleus staining, and in cell numbers. Conclusion: The optimal combination of hormones and EAAs increased BMEC proliferation by enhancing cell cycle progression in the S and G/2M phases. Our findings indicate that optimizing hormone and amino acid levels has the potential to enhance milk production, both in cell culture settings by promoting increased cell numbers, and in dairy cows by regulating feed intake.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.197-202
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• 1997

Effect of arabinose on xylitol production from xylose by Candida parapsilosis KFCC 10875 was investigated at the different concentrations of arabinose. When the arabinose was added in xylose medium, the cell growth increased and the final cell concentration was maximum at 10 g/l arabinose. The consumption rate of arabinose was greatly lower than those of xylose and arabinose. Above 10 g/l arabinose, it was not completely consumed and then remained in the medium during xylitol fermentation. Estimated cell mass obtained from arabinose increased with increasing consumed arabinose. As arabinose concentration was increased, xylitol production decreased but ethanol production increased. The inhibitory effect of ethanol, a major by-product, on xylitol production was also studied. As the ethanol concentration added increased, xylitol production decreased. When cells were inoculated in a xylose medium after removing ethanol, xylitol production was not inhibited. This results suggested that the inhibition of xylitol production resulted from ethanol which was formed by adding arabinose. It was also interesting that total products(xylitol and ethanol) yield was constant regardless of the arabinose concentration. This result suggested that the total amount of products such as xylitol and ethanol from xylose was constant regardless of the arabinose concentration and arabinose shifted the carbon flow from xylitol to ethanol.

• KSBB Journal
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• 제25권2호
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• pp.193-198
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• 2010

Taxus wallichiana Zucc. 현탁세포배양에서 항암제 paclitaxel 생합성 연구를 수행하였다. 현탁세포배양에서 exponential phase의 최고 생장 속도는 0.14 $day^{-1}$이었으며 배가시간이 4.96 day이었다. Paclitaxel은 배양초기에 생성되지 않고 exponential phase 말기부터 서서히 생성되어 stationary phase에서도 이어지며 declined phase에서 최고치를 나타내었다. Paclitaxel의 생산을 촉진하기 위하여 elicitor를 사용하였다. 여러 elicitor 중 jasmonic acid와 cellulase가 paclitaxel의 생성을 증가시켰다. Elicitor는 접종과 동시에 투여했을 때 가장 효과가 좋았으며, jasmonic acid와 cellulase의 combination으로 효과가 더욱 증진했는데 각각 투여할 때보다 1.8배 및 3.1배 paclitaxel 생성이 증가되었다.

• Preventive Nutrition and Food Science
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• 제11권4호
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• pp.273-277
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• 2006

Recently, Rhei Rhizoma extracts (RRE) have begun to receive more attention as potential biological response modifiers. In the present study, we studied the antiproliferative effect of RRE on tumor cells and the effect of RRE on macrophage function. A variety of tumor cells and macrophages were treated with RRE at various concentrations. The effect of RRE on cell proliferation was measured by MTT assay and the effect of RRE on the production of nitric oxide (NO) was determined in the macrophage-like cell lines Raw264.7, C6 and peritoneal macrophages (pMQ). RRE inhibited the growth of tumor cells (e.g., B16, HOS). However, the effects of RRE on the production of NO varied with macrophage types. RRE had no effect on C6 cell growth and slightly increased the growth of Raw264.7 cells. In addition, treatment of normal pMQ with RRE enhanced NO production in a concentration-dependent manner, whereas RRE suppressed NO production at $50\;{\mu}g/mL$ in both Raw264.7 and C6 cells. However, RRE suppressed NO production in LPS/IFN-$\gamma$-stimulated C6 cells. Overall, these results suggest that RRE elicits an antiproliferative property and differentially modulates NO production in various macrophages, and have a potential for therapeutic application.

• 통합자연과학논문집
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• 제1권1호
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• pp.41-46
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• 2008

The aim of this study is to evaluate the effects essential oil from Lavendular officinalis on the production of UVB-irradiated-induced nitric oxide(NO), in vivo and in vitro. NO is a recently discovered mediator of cell communication involved in a variety of physiological and pathophysiological processes. This enzyme is present in various tissues including smooth muscle cells and macrophages and take part in several immunopathological process. In vitro, the cytotoxicity and cell viability of aroma oil was evaluated by the MTT assay in the concentration of 0.01, 0.05, 0.1%. And, the effect of aroma oil was investigated to production of NO in human fibroblast cells line CCD-986sk ($2{\times}10^5$ cell/well) after UVB-irradiation with aroma oil (0.01, 0.1, and 1%). The result showed that aroma oil did not affected the production of NO. In vivo, it was investigated to production of NO after UVB- irradiation with aroma oil. The experimental groups were divided into four groups. Aroma oil was stimulated the production of NO by itself. As the results, all of the in vitro and in vivo, aroma oil were affected production of NO by dependent the concentration-manners.

• 한국축산식품학회지
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• 제35권4호
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• pp.541-550
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• 2015

The purpose of this study was to investigate the effect of lactic acid bacteria (LAB) cell wall extract on the proliferation and cytokine production of immune cells to select suitable probiotics for space food. Ten strains of LAB (Lactobacillus bulgaricus, L. paracasei, L. casei, L. acidophilus, L. plantarum, L. delbruekii, Lactococcus lactis, Streptococcus thermophilus, Bifidobacterium breve, and Pedicoccus pentosaceus) were sub-cultured and further cultured for 3 d to reach 7-10 Log colony-forming units (CFU)/mL prior to cell wall extractions. All LAB cell wall extracts failed to inhibit the proliferation of BALB/c mouse splenocytes or mesenteric lymphocytes. Most LAB cell wall extracts except those of L. plantarum and L. delbrueckii induced the proliferation of both immune cells at tested concentrations. In addition, the production of T_H1 cytokine (IFN-γ) rather than that of T_H2 cytokine (IL-4) was enhanced by LAB cell wall extracts. Of ten LAB extracts, four (from L. acidophilus, L. bulgaricus, L. casei, and S. thermophiles) promoted both cell proliferating and T_H1 cytokine production. These results suggested that these LAB could be used as probiotics to maintain immunity and homeostasis for astronauts in extreme space environment and for general people in normal life.

• KSBB Journal
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• 제13권3호
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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.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.261-264
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• 2002

CHO (Chinese hamster ovary) cells were transfected with plasmids containing both cis-acting HRE (hypoxia response element) and CMV-promoter that controls tissue-type plasminogen activator (t-PA). CHO cells with HRE produced 16.2 fold higher t-PA concentration than CHO cells without HRE. It was noted that hypoxia strongly induced CHO cell apoptosis. which resulted in decrease of cell viability and protein production. In this study. by introducing Bcl-2, anti-apoptotic gene, we tried to recover cell viability and increase the protein production. When batch culture of both control cells without transfection of Bcl-2 and cells transfected with Bcl-2 were performed in the absence of CoCl ι hypoxia mimic condition. the cells with Bcl-2 were effected specific cell growth rates, maximum cell density. Immunoblotting assay showed Bcl-2 was recombinant with HRE dependent t- P A expression cassette, and their expression level was depended on hypoxia. By introducing Bcl-2, both cell viability and maximum cell density could be increased.

• BMB Reports
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• 제52권5호
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• pp.295-303
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• 2019

Breakthroughs in stem cell technology have contributed to disease modeling and drug screening via organoid technology. Organoid are defined as three-dimensional cellular aggregations derived from adult tissues or stem cells. They recapitulate the intricate pattern and functionality of the original tissue. Insulin is secreted mainly by the pancreatic ${\beta}$ cells. Large-scale production of insulin-secreting ${\beta}$ cells is crucial for diabetes therapy. Here, we provide a brief overview of organoids and focus on recent advances in protocols for the generation of pancreatic islet organoids from pancreatic tissue or pluripotent stem cells for insulin secretion. The feasibility and limitations of organoid cultures derived from stem cells for insulin production will be described. As the pancreas and gut share the same embryological origin and produce insulin, we will also discuss the possible application of gut organoids for diabetes therapy. Better understanding of the challenges associated with the current protocols for organoid culture facilitates development of scalable organoid cultures for applications in biomedicine.

• KSBB Journal
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• 제14권4호
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• pp.445-451
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• 1999

E. coli TGI/pDG7 $\alpha$ 와 빵효모의 혼합세포계에 의한 글루타치온 생산을 Aerated Slurry Bioreactor에서 수행하였다. 글루타치온 생산 효소는 34시간 동안 안정성을 보였으며, 글루타치온 수율은 4.6 mM을 유지함을 보였다. 고농도의 혼합세포를 이용한 Aerated Slurry Bioreactor에서 기질의 공급속도가 글루타치온 생산에 미치는 영향을 조사하였다. 세포가 회수되어 기질공급속도 5.2 mL/hr에서 글루타치온의 생산농도는 32시간에서 현탁반응보다 낮아짐을 보였고, 반면 세포가 회수되지 않은 기질공급속도 5.2 mL/hr에서는 22시간 동안 유지되었고 2.6 mL/hr에서는 42시간 동안 유지되었다. 글루타치온의 생산성은 기질공급속도가 10.4 mL/hr에서 25.7 mg/g wet $cell{\cdot}hr$이었으나, 2.6 mL/hr에서는 1.65 mg/g wet $cell{\cdot}hr$이었고 5.2mL/hr에서는 5.28 mg/g wet $cell{\cdot}hr$이었다. 계면활성제(Tween 80)을 사용한 경우, 글루타치온 생산시간을 25시간에서 45시간으로 연장되었다. 글루타치온 생산있어 산소가 세포계의 효소에 산소에 의해 영향을 미칠 것으로 판단되었고, 공기를 질소로 교체하여 글루타치온 생산시간을 40시간으로 연장시켰다.