scholarly journals Expression of Human Erythropoietin Containing 2 Additional N-Link in CHO-K1 Cells under Different Culture Conditions

2019 ◽  
Vol 10 (1) ◽  
pp. 7
Author(s):  
Adi Santoso ◽  
Larasati Larasati ◽  
Arizah Kusumawati ◽  
Popi Hadi Wisnuwardhani ◽  
Ratih Asma Ningrum ◽  
...  
Keyword(s):  
Cell Culture ◽  
Protein Expression ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Culture Conditions ◽  
Expression Level ◽  
Seeding Density ◽  
Cell Culture Media ◽  
Human Erythropoietin ◽  
Fetal Bovine

Human erythropoietin (hEPO) is a glycoprotein that regulates the formation of erythrocytes and mainly used in anemia patients. Previously, we have reported the expression of modified human EPO with 2 additional N-linked in mammalian cell CHO-K1. The aim of this current research was to study the optimum condition for modified recombinant hEPO (rhEPO) production in CHO-K1. To do this, several parameters of culture conditions were applied including antibiotic concentrations, seeding densities, time of incubations, fetal bovine serum (FBS) concentrations and cell culture media. The result showed that the presence of antibiotic G418 improved the expression level with the highest was at 1% of concentration. Meanwhile, seeding density of 2–3x105 cells/6 cm dish and seven day of incubation time were the best condition for rhEPO protein expression. From five different combination media used, F12 medium with 10% FBS gave the highest expression of rhEPO protein. From this study was also found that at passage 16 the expression level was still increasing proving that the clone expressing the protein of our interest is promisingly stable.Keywords : EPO, erythropoietin, protein expression, CHO-K1, optimation

scholarly journals A plea to reduce or replace fetal bovine serum in cell culture media

2013 ◽  
Vol 65 (5) ◽  
pp. 791-793 ◽  
Author(s):  
Gerhard Gstraunthaler ◽  
Toni Lindl ◽  
Jan van der Valk
Keyword(s):  
Cell Culture ◽  
Bovine Serum ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Cell Culture Media ◽  
Fetal Bovine

Optimization of chemically defined cell culture media – Replacing fetal bovine serum in mammalian in vitro methods

2010 ◽  
Vol 24 (4) ◽  
pp. 1053-1063 ◽  
Author(s):  
J. van der Valk ◽  
D. Brunner ◽  
K. De Smet ◽  
Å. Fex Svenningsen ◽  
P. Honegger ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bovine Serum ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Cell Culture Media ◽  
In Vitro Methods ◽  
Fetal Bovine

Fetal bovine serum—a cell culture dilemma

Science ◽  
2022 ◽  
Vol 375 (6577) ◽  
pp. 143-144
Author(s):  
Jan van der Valk
Keyword(s):  
Cell Culture ◽  
Bovine Serum ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Cell Culture Media ◽  
A Cell ◽  
Fetal Bovine

Ethical and possible reproducibility issues arise when using fetal bovine serum in cell culture media

scholarly journals Application of DNA Quadruplex Hydrogels Prepared from Polyethylene Glycol-Oligodeoxynucleotide Conjugates to Cell Culture Media

Polymers ◽  
2019 ◽  
Vol 11 (10) ◽  
pp. 1607
Author(s):  
Shizuma Tanaka ◽  
Shinsuke Yukami ◽  
Yuhei Hachiro ◽  
Yuichi Ohya ◽  
Akinori Kuzuya
Keyword(s):  
Cell Culture ◽  
High Efficiency ◽  
Culture Media ◽  
Culture Conditions ◽  
Cell Culture Media ◽  
G Quadruplex ◽  
L929 Fibroblast ◽  
Dna Quadruplex ◽  
Quadruplex Formation ◽  
Poly Ethylene

Application of Na+-responsive DNA quadruplex hydrogels, which utilize G-quadruplexes as crosslinking points of poly(ethylene glycol) (PEG) network as cell culture substrate, has been examined. PEG-oligodeoxynucleotide (ODN) conjugate, in which four deoxyguanosine (dG4) residues are tethered to both ends of PEG, was prepared by modified high-efficiency liquid phase (HELP) synthesis of oligonucleotides and used as the macromonomer. When mixed with equal volume of cell culture media, the solution of PEG-ODN turned into stiff hydrogel (G-quadruplex hydrogel) as the result of G-quadruplex formation by the dG4 segments in the presence of Na+. PEG-ODN itself did not show cytotoxicity and the resulting hydrogel was stable enough under cell culture conditions. However, L929 fibroblast cells cultured in G-quadruplex hydrogel remained spherical for a week, yet alive, without proliferation. The cells gradually sedimented through the gel day by day, probably due to the reversible nature of G-quadruplex formation and the resulting slow rearrangement of the macromonomers. Once they reached the bottom glass surface, the cells started to spread and proliferate.

scholarly journals Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

2013 ◽  
Vol 33 (suppl 1) ◽  
pp. 20-24 ◽  
Author(s):  
Gesiane Ribeiro ◽  
Cristina O. Massoco ◽  
José Corrêa de Lacerda Neto
Keyword(s):  
Cell Culture ◽  
Bone Marrow ◽  
Adipose Tissue ◽  
Culture Media ◽  
Stromal Vascular Fraction ◽  
Bone Marrow Aspiration ◽  
Cell Culture Media ◽  
Colony Forming Units ◽  
Fetal Bovine ◽  
Equine Bone

The objective of this study was to evaluate the culture of equine bone marrow mononuclear fraction and adipose tissue - derived stromal vascular fraction cells in two different cell culture media. Five adult horses were submitted to bone marrow aspiration from the sternum, and then from the adipose tissue of the gluteal region near the base of the tail. Mononuclear fraction and stromal vascular fraction were isolated from the samples and cultivated in DMEM medium supplemented with 10% fetal bovine serum or in AIM-V medium. The cultures were observed once a week with an inverted microscope, to perform a qualitative analysis of the morphology of the cells as well as the general appearance of the cell culture. Colony-forming units (CFU) were counted on days 5, 15 and 25 of cell culture. During the first week of culture, differences were observed between the samples from the same source maintained in different culture media. The number of colonies was significantly higher in samples of bone marrow in relation to samples of adipose tissue.

scholarly journals The Cell Culture Medium Affects Growth, Phenotype Expression and the Response to Selenium Cytotoxicity in A549 and HepG2 Cells

Antioxidants ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 130 ◽  
Author(s):  
Lisa Arodin Selenius ◽  
Marita Wallenberg Lundgren ◽  
Rim Jawad ◽  
Olof Danielsson ◽  
Mikael Björnstedt
Keyword(s):  
Cell Culture ◽  
Cell Growth ◽  
Culture Medium ◽  
Culture Media ◽  
A549 Cells ◽  
Culture Conditions ◽  
Cell Culture Medium ◽  
Cell Culture Media ◽  
Toxic Response ◽  
Selenium Compounds

Selenium compounds influence cell growth and are highly interesting candidate compounds for cancer chemotherapy. Over decades an extensive number of publications have reported highly efficient growth inhibitory effects with a number of suggested mechanisms f especially for redox-active selenium compounds. However, the studies are difficult to compare due to a high degree of variations in half-maximal inhibitor concentration (IC50) dependent on cultivation conditions and methods to assess cell viability. Among other factors, the variability in culture conditions may affect the experimental outcome. To address this, we have compared the maintenance effects of four commonly used cell culture media on two cell lines, A549 and HepG2, evaluated by the toxic response to selenite and seleno-methylselenocysteine, cell growth and redox homeostasis. We found that the composition of the cell culture media greatly affected cell growth and sensitivity to selenium cytotoxicity. We also provided evidence for change of phenotype in A549 cells when maintained under different culture conditions, demonstrated by changes in cytokeratin 18 (CK18) and vimentin expression. In conclusion, our results have shown the importance of defining the cell culture medium used when comparing results from different studies.

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