The cell culture medium ? a functional extracellular compartment of suspension-cultured cells

1994 ◽  
Vol 38 (2-3) ◽  
pp. 307-319 ◽  
Author(s):  
Michael Wink
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Cultured Cells ◽  
Cell Culture Medium ◽  
Suspension Cultured Cells ◽  
Extracellular Compartment

Endothelial cell cystine uptake and glutathione increase with N,N-bis(2-chloroethyl)-N-nitrosourea exposure

1992 ◽  
Vol 262 (3) ◽  
pp. L301-L304 ◽  
Author(s):  
S. M. Deneke ◽  
R. A. Lawrence ◽  
S. G. Jenkinson
Keyword(s):  
Cell Culture ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Culture Medium ◽  
Cultured Cells ◽  
Potent Inhibitor ◽  
Cell Types ◽  
Cell Culture Medium ◽  
Intracellular Accumulation ◽  
Pulmonary Endothelial Cells

Glutathione (gamma-glutamylcysteinylglycine, GSH) is an important cellular antioxidant. In typical cultured cell preparations GSH synthesis is limited by the availability of intracellular cysteine. Because extracellular cystine is the chief source of intracellular cysteine in cultured cells, increasing cystine transport can result in increased intracellular GSH. Depletion of GSH or exposure to oxidants has been shown to stimulate cystine transport in bovine pulmonary endothelial cells and other cell types. BCNU [N,N-bis(2-chloroethyl)-N-nitrosourea] is a potent inhibitor of glutathione reductase (GSSG-Red). We examined the effects of BCNU on cystine uptake by bovine pulmonary artery endothelial cells (BPAEC). We hypothesized that blocking GSSG-Red could result in increased cellular uptake of cystine to replenish decreases in GSH caused by oxidation. Levels of BCNU between 0.005 and 0.05 mM added to the cell culture medium inhibited GSSG-Red at 2, 4, and 24 h after addition. BCNU treatment resulted in concentration-dependent increases in both cystine uptake and GSH levels after 24 h of exposure. The increases in uptake were specific for cystine and glutamate and were sodium independent, suggesting induction of a xc(-)-like transport system. No intracellular accumulation of GSSG was measured nor was any significant depletion of GSH noted at any time of BCNU exposure.

The Use of Artemia Salina in Toxicity Testing

1992 ◽  
Vol 20 (2) ◽  
pp. 297-301 ◽  
Author(s):  
Lillemor Lewan ◽  
Marianne Andersson ◽  
Paloma Morales-Gomez
Keyword(s):  
Cell Culture ◽  
Mammalian Cells ◽  
Culture Medium ◽  
Cultured Cells ◽  
Sea Water ◽  
Toxicity Testing ◽  
Artemia Salina ◽  
Good Survival ◽  
Cell Culture Medium ◽  
A Cell

This study shows that the Artemia assay, which is usually performed by incubation for a 24-hour period in artificial sea water, can also be performed in phosphate buffered saline (PBS) at pH 7.2, or in a cell culture medium, both of which are used in toxicity assays employing mammalian cells. Thus, by using the equivalent media for incubation, toxic effects in the Artemia assay and toxic mechanisms can more accurately be compared with results obtained in mammalian cell toxicity. Survival of control animals is good for 72 hours, provided that infection can be avoided. A pH greater than 6 is essential for good survival of Artemia Salina, and a pH greater than 10.5 should be avoided. Because of the risk of infection at low saline concentrations, a decreased incubation time of 16 or 12 hours is recommended. The lethal concentrations (LC10 and LC50) in the 24-hour Artemia assay of the first ten chemicals in the MEIC programme were measured in PBS, and the results compared with those from a previous study of the effects in a 10-minute acute ATP leakage assay, specifically indicating lesions in the cell membrane. The EC10 and EC50 values for the alcohols in the Artemia assay were 35–75% of the corresponding values in the ATP leakage assay. For paracetamol and amitriptyline, the EC10 and EC50 values in the Artemia assay were 2–16% of the corresponding values in the ATP-leakage assay. The greatly increased toxicity of the two compounds in the animals may be related to systemic effects. FeSO4 was very toxic to Artemia salina at a concentration of lug/ml, but it did not cause leakage of ATP from cultured cells, even at a concentration of 8,000μg/ml, showing that widely different mechanisms of interaction with FeSO4 are measured by the two assays. A lower toxicity of polygodial, a sesquiterpenoid unsaturated dialdehyde, in cell culture medium, was obvious in the Artemia assay. Thus, some factors in cell culture medium must, by interacting with the toxic molecule, protect the animals against the toxicity of polygodial, as was previously found in cultured cells.

3-iodothyronamine (T1AM) is taken up and rapidly metabolized in cell culture medium only at low concentration

2020 ◽  
Author(s):  
Federica Saponaro ◽  
Marco Borsò ◽  
Sara Verlotta ◽  
Lavinia Bandini ◽  
Alessandro Saba ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Low Concentration

Effect of Atmospheric-Pressure Helium Plasma Jet on Cell Culture Medium

2013 ◽  
Vol 133 (5) ◽  
pp. 278-285
Author(s):  
Norimitsu Takamura ◽  
Douyan Wang ◽  
Takao Satoh ◽  
Takao Namihira ◽  
Hisato Saitoh ◽  
...  
Keyword(s):  
Cell Culture ◽  
Atmospheric Pressure ◽  
Culture Medium ◽  
Plasma Jet ◽  
Helium Plasma ◽  
Cell Culture Medium

scholarly journals Defined MSC exosome with high yield and purity to improve regenerative activity

2021 ◽  
Vol 12 ◽  
pp. 204173142110086
Author(s):  
Jun Yong Kim ◽  
Won-Kyu Rhim ◽  
Yong-In Yoo ◽  
Da-Seul Kim ◽  
Kyoung-Won Ko ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Density Lipoprotein ◽  
High Yield ◽  
Human Umbilical Cord ◽  
Cell Culture Medium ◽  
Human Coronary Artery ◽  
Isolation Methods ◽  
Regenerative Activity ◽  
Yield And Purity

Exosomes derived from mesenchymal stem cells (MSCs) have been studied as vital components of regenerative medicine. Typically, various isolation methods of exosomes from cell culture medium have been developed to increase the isolation yield of exosomes. Moreover, the exosome-depletion process of serum has been considered to result in clinically active and highly purified exosomes from the cell culture medium. Our aim was to compare isolation methods, ultracentrifuge (UC)-based conventional method, and tangential flow filtration (TFF) system-based method for separation with high yield, and the bioactivity of the exosome according to the purity of MSC-derived exosome was determined by the ratio of Fetal bovine serum (FBS)-derived exosome to MSC-derived exosome depending on exosome depletion processes of FBS. The TFF-based isolation yield of exosome derived from human umbilical cord MSC (UCMSC) increased two orders (92.5 times) compared to UC-based isolation method. Moreover, by optimizing the process of depleting FBS-derived exosome, the purity of UCMSC-derived exosome, evaluated using the expression level of MSC exosome surface marker (CD73), was about 15.6 times enhanced and the concentration of low-density lipoprotein-cholesterol (LDL-c), known as impurities resulting from FBS, proved to be negligibly detected. The wound healing and angiogenic effects of highly purified UCMSC-derived exosomes were improved about 23.1% and 71.4%, respectively, with human coronary artery endothelial cells (HCAEC). It suggests that the defined MSC exosome with high yield and purity could increase regenerative activity.

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