• Title/Summary/Keyword: mammalian cells

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Effect of Microsparged Aeration on Oxygen Transfer Rate and Cell Viability in Mammalian Cell Culture Bioreactor (동물 세포 반응기에서의 초미세 통기법이 산소 전달 속도와 세포 생존율에 미치는 영향)

  • 김정모;장건희;최춘순;김정회
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.240-247
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    • 2001
  • The effect of microsparged aeration in mammalian cell bioreactor on the oxygen transfer rate and cell viability was studied. The microspargers with differ- ent micron-sized pores were used to supply oxygen to the medium. The oxygen transfer coefficients (k$_{L}$a) measured in the bioreactor were markedly increased, which is due to the increase of the contacting area between air bubbles and liquid medium when the pore size of microsparger decreases. When the impellers of two different types (square-pitch marine impeller and $45^{\circ}$ pitched flat blade impeller) were used for agitation, the k$_{L}$a values were slightly higher with the marine impeller than with the blade impeller. The detrimental effect of direct gas sparging with microsparger on mammalian cells was investigated in bubble columns with various air flow rates and different pore sized microspargers. The first-order cell death rate constant ($k_{d}$ /7) was shown to be directly proportional to the air flow rate and inversely proportional to the pore size. During the cultivation of hybridoma cells using microsparger with the pore size of $0.57\mu$m in the mammalian cell culture bioreactor, the continuous sparging caused the cell death and suppressed the cell growth. However, cells grew normally and cell viability was maintained above 90% in the logarithmic phase when the air was intermittently sparked in order to maintain the dissolved oxygen level above 20%.

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Apoptosis-Inducing Activity of HPLC Fraction from Voacanga globosa (Blanco) Merr. on the Human Colon Carcinoma Cell Line, HCT116

  • Acebedo, Alvin Resultay;Amor, Evangeline Cancio;Jacinto, Sonia Donaldo
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.617-622
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    • 2014
  • Voacanga globosa (Blanco), a plant endemic to the Philippines, is traditionally used especially by indigenous people of Bataan in the treatment of ulcers, wounds and tumorous growths. This study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of HPLC fractions from leaves on HCT116 human colon carcinoma and A549 human lung carcinoma cell lines. Ethanolic extraction was performed on V globosa leaves followed by hexane and ethyl acetate partitioning. Silica gel column chromatography and high performance liquid chromatography (HPLC) produced MP1, MP2 and MP3 fractions. Cytotoxic activity of the fractions was determined through MTT assay against the cancer cell lines HCT116 and A549 and the non-cancer AA8 Chinese hamster ovarian cell line. Pro-apoptotic activities of the most active fractions were further assessed through DAPI staining, TUNEL assay and JC-1 mitochondrial membrane potential assay with HCT116 cells. While the MPI fraction exerted no significant activity against all cell lines tested, MP2 and MP3 fractions demonstrated high toxicity against HCT116 and A549 cells. The MP3 fraction induced formation of apoptotic bodies, condensed DNA and other morphological changes consistent with apoptosis of HCT116 cells and TUNEL assay showed significant increase in DNA fragmentation over time. In these cells, the MP3 fraction also induced mitochondrial membrane destabilization, which is generally associated with the beginning of apoptosis. Phytochemical analysis demonstrated the presence only of saponins and terpenoids in the MP3 fraction. The results indicate that the MP3 fraction exerts cytotoxic activity on HCT116 cells via induction of apoptosis triggered by loss of mitochondrial membrane potential crucial for cell survival.

Kinetics of Cultivating Large Quantities of Mammalian Cells (tPA생산을 위한 동물 세포 배양에 관한 동력학적 연구)

  • 이현용
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.282-286
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    • 1988
  • Growth kinetic parameters for mass cultivation of Chinese Hamster Ovary (CHO) cells are estimated by measuring oxygen uptake rates. It Is found that there is strong correlation between cell growth and oxygen consumption, showing that correlation factor is 0.83. Derived linear model predicts actual cell density very well. It tells that oxygen uptake rate can play important role in indirectly measuring cell density when conventional method of estimating cell density is no longer meaningful due to heavy cell clumpings. Cell yield per oxygen consumption, $Y_{\chi}o$ and mass transfer coefficient for oxygen, Ka are also estimated as 1.26$\times$10$^4$cells/mmole $O_2$ consumed and 1.01/h, respectively. Average specific growth rate over all runs is 2.891/day for CHO cells with producting 2 grams of tPA per day under continuous perfusion operations.

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Genome editing of immune cells using CRISPR/Cas9

  • Kim, Segi;Hupperetz, Cedric;Lim, Seongjoon;Kim, Chan Hyuk
    • BMB Reports
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    • v.54 no.1
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    • pp.59-69
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    • 2021
  • The ability to read, write, and edit genomic information in living organisms can have a profound impact on research, health, economic, and environmental issues. The CRISPR/Cas system, recently discovered as an adaptive immune system in prokaryotes, has revolutionized the ease and throughput of genome editing in mammalian cells and has proved itself indispensable to the engineering of immune cells and identification of novel immune mechanisms. In this review, we summarize the CRISPR/Cas9 system and the history of its discovery and optimization. We then focus on engineering T cells and other types of immune cells, with emphasis on therapeutic applications. Last, we describe the different modifications of Cas9 and their recent applications in the genome-wide screening of immune cells.

Biological Roles of Alternative Autophagy

  • Shimizu, Shigeomi
    • Molecules and Cells
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    • v.41 no.1
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    • pp.50-54
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    • 2018
  • Atg5 and Atg7 have long been considered as essential molecules for autophagy. However, we found that cells lacking these molecules still form autophagic vacuoles and perform autophagic protein degradation when subjected to certain stressors. During this unconventional autophagy pathway, autophagosomes appeared to be generated in a Rab9-dependent manner by the fusion of vesicles derived from the trans-Golgi and late endosomes. Therefore, mammalian autophagy can occur via at least two different pathways; the Atg5/Atg7-dependent conventional pathway and an Atg5/Atg7-independent alternative pathway.

ZR-75-1 human breast cancer cells to study the mechanism of action of PAHs

  • Min, Kyung-Nan;Sheen, Yhun-Yhong
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.286.1-286.1
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    • 2002
  • Recent industrial society has human widely exposed to PAHs that are comming from the incomplete combustion of organic material as widespread environmental contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. PAHs in the mammalian cells affect CYP 1A1 gene expression as well as other phase II drug metabolizing enzymes as UDPGT, NMOR etc. The mechanism of action of PAHs has been studied extensively, however it is not clear how PAHs turn on CYPIAI in human breast cancer. (omitted)

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A new strategy for high productivity of Erythropoietin in CHO cell by introducing urea cycle enzymes

  • Lee, Yun-Jeong;Chang, Kern-Hee;Kim, Jung-Hoe;Kim, Hong-Jin
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.338.1-338.1
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    • 2002
  • The efficient Erythropoietin(EPO)-expression system in mammalian cells is required for massive production for therapeutic use. Ammonium ion is a major problem in the production of useful proteins by cultured animal cells and therefore it is of importance to devise a system by which a high productivity of human therapeutic recombinant protein can be maintained or enhanced under low ammonium concentration. (omitted)

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