Trypanosoma musculi: In vitro cultivation of blood forms in cell culture media

1977 ◽  
Vol 7 (2) ◽  
pp. 109-111 ◽  
Author(s):  
P. Viens ◽  
M.C. Lajeunesse ◽  
R. Richards ◽  
G.A.T. Targett
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
In Vitro Cultivation ◽  
Cell Culture Media

scholarly journals Impact of serum in cell culture media on in vitro lactate dehydrogenase (LDH) release determination

2017 ◽  
Vol 3 (1) ◽  
pp. 9-13 ◽  
Author(s):  
Bernhard Hiebl ◽  
Sinem Peters ◽  
Ole Gemeinhardt ◽  
Stefan M. Niehues ◽  
Friedrich Jung
Keyword(s):  
Cell Culture ◽  
Lactate Dehydrogenase ◽  
Culture Media ◽  
Cell Culture Media ◽  
Ldh Release

scholarly journals EFFECT OF IN VITRO CULTIVATION ON THE PATHOGENICITY OF WEST NILE VIRUS

1946 ◽  
Vol 84 (2) ◽  
pp. 181-190 ◽  
Author(s):  
Hilary Koprowski ◽  
Edwin H. Lennette
Keyword(s):  
West Nile Virus ◽  
Culture Media ◽  
Lethal Effect ◽  
In Vitro Cultivation ◽  
Suspended Cell ◽  
West Nile ◽  
Cell Culture Media ◽  
Full Capacity ◽  
Old Mice

The West Nile virus was cultivated in suspended cell culture media employing several different tissue components, and it has been observed to survive in culture for at least 32 days. Continued propagation of the virus in vitro resulted in a change in its pathogenicity. The change lay in a marked reduction or a complete loss of the ability of the virus to produce fatal infections in mice and in hamsters on peripheral inoculation, although there was no obvious simultaneous alteration in the lethal effect of the virus by the cerebral route. In mice, the extent to which invasiveness was lost depended upon the passage level of the virus and the age of the test animals. The younger (and more susceptible) the mice, the greater the number of passages which was required to diminish the virulence of the virus by peripheral routes; after 68 passages, the virus still retained its full capacity to kill 3-day-old mice, while its ability to kill 8-day-old mice was reduced and its ability to kill mice 14 or more days of age was essentially abolished. How soon the loss of pathogenicity occurs in hamsters has not been determined. Prolonged cultivation rendered the virus avirulent for hamsters by the intraperitoneal route.

Evidence for a Metabolic Stem Cell Niche.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4046-4046 ◽  
Author(s):  
Michael Cross ◽  
Rudiger Alt ◽  
Lydia Schnapke-Hille ◽  
Thomas Riemer ◽  
Dietger Niederwieser
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Stem Cell ◽  
Stem Cell Niche ◽  
Culture Media ◽  
Self Renewal ◽  
Cell Culture Media ◽  
Hematopoietic Stem ◽  
Cell Niche

Abstract The hematopoietic stem cell niche presents a localised environment supporting the balanced maintenance, self-renewal and occasional expansion of the stem cell pool. These options are widely assumed to be regulated exclusively by signalling from specific combinations of stroma-bound or soluble ligands. However, a consideration of the rare conditions under which absolute numbers of stem cells increase in vivo as well as the selective pressures acting on regenerative systems during evolution has led us to propose a metabolic component to the stem cell niche which serves to limit cumulative damage, to avoid the selection of potentially oncogenic mutations and to tie symmetric division to slow proliferation. This would mean that traditional cell culture media based on “systemic” substrates such as glucose and glutamine may actively prevent the symmetric amplification of high quality stem cells, offering a possible explanation for the limited success in this area to date. To investigate this possibility, we have examined the effects of range of carbon and energy sources on the proliferation and maintenance of stem and progenitor cells. Our strategy is to screen a wide variety of culture conditions using murine FDCPmix cells, which are non-tumorigenic but have an innate tendency to amplify symmetrically in the presence of IL-3, and then to test key observations in human UCB CD133+ cells provided with SCF, TPO and FLT-3L. In both cell systems, we do indeed find an unusually low requirement for the systemic substrates glucose and glutamine normally included as major energy and carbon sources in cell culture media. Reducing glucose reduces the yield of committed cells from CD133+ cultures without affecting the accumulation of CD133+CD34+cKit+ progenitors. When provided with alternative substrates more likely to reflect a “niche” type environment, FDCPmix cells can be maintained for long periods in media containing only the trace levels of glucose or glutamine derived from dialysed serum, and show improved self-renewal under these conditions. We have also found that raising osmolarity reduces glucose dependence and simultaneously favours the maintenance both of self-renewing CFU (FDCPmix culture) and of CAFCweek13 (CD133+ culture). In parallel, the use of NMR and mass spectrometry techniques to profile intracellular metabolites in self-renewing and differentiating FDCPmix cells reveals a shift in the metabolite balance indicating reduced glycolysis in the early cells. Taken together, these results suggest that hematopoietic stem cells do indeed have remarkable metabolic characteristics consistent with adaptation to a metabolically limiting niche environment. It may therefore be necessary to identify niche substrates and to combine these with the relevant signalling environment in vitro in order to effectively increase stem cell numbers for research, stem cell transplantation and tissue engineering applications.

scholarly journals Surpassing light-induced cell damage in vitro with novel cell culture media

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
John H. Stockley ◽  
Kimberley Evans ◽  
Moritz Matthey ◽  
Katrin Volbracht ◽  
Sylvia Agathou ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Cell Damage ◽  
Cell Culture Media

scholarly journals In Vitro Bioavailability of the Hydrocarbon Fractions of Dimethyl Sulfoxide Extracts of Petroleum Substances

2020 ◽  
Vol 174 (2) ◽  
pp. 168-177 ◽  
Author(s):  
Yu-Syuan Luo ◽  
Kyle C Ferguson ◽  
Ivan Rusyn ◽  
Weihsueh A Chiu
Keyword(s):  
Cell Culture ◽  
Protein Binding ◽  
Aromatic Compounds ◽  
Culture Media ◽  
In Vitro Testing ◽  
Cell Culture Media ◽  
Chemical Profiles ◽  
In Vivo Extrapolation

Abstract Determining the in vitro bioavailable concentration is a critical, yet unmet need to refine in vitro-to-in vivo extrapolation for unknown or variable composition, complex reaction product or biological material (UVCB) substances. UVCBs such as petroleum substances are commonly subjected to dimethyl sulfoxide (DMSO) extraction in order to retrieve the bioactive polycyclic aromatic compound (PAC) portion for in vitro testing. In addition to DMSO extraction, protein binding in cell culture media and dilution can all influence in vitro bioavailable concentrations of aliphatic and aromatic compounds in petroleum substances. However, these in vitro factors have not been fully characterized. In this study, we aimed to fill in these data gaps by characterizing the effects of these processes using both a defined mixture of analytical standards containing aliphatic and aromatic hydrocarbons, as well as 4 refined petroleum products as prototypical examples of UVCBs. Each substance was extracted with DMSO, and the protein binding in cell culture media was measured by using solid-phase microextraction. Semiquantitative analysis for aliphatic and aromatic compounds was achieved via gas chromatography-mass spectrometry. Our results showed that DMSO selectively extracted PACs from test substances, and that chemical profiles of PACs across molecular classes remained consistent after extraction. With respect to protein binding, chemical profiles were retained at a lower dilution (higher concentration), but a greater dilution factor (ie, lower concentration) resulted in higher protein binding in cell medium, which in turn altered the ultimate chemical profile of bioavailable PACs. Overall, this case study demonstrates that extraction procedures, protein binding in cell culture media, and dilution factors prior to in vitro testing can all contribute to determining the final bioavailable concentrations of bioactive constituents of UVCBs in vitro. Thus, in vitro-to-in vivo extrapolation for UVCBs may require greater attention to the concentration-dependent and compound-specific differences in recovery and bioavailability.

In vitro cytocompatibility testing of oxidative degradation products

2021 ◽  
pp. 088391152110031
Author(s):  
Scott M Herting ◽  
Mary Beth B Monroe ◽  
Andrew C Weems ◽  
Sam T Briggs ◽  
Grace K Fletcher ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Cell Culture ◽  
Culture Media ◽  
Degradation Products ◽  
Oxidative Degradation ◽  
Cell Culture Media ◽  
Novel Approach ◽  
Gene And Protein Expression

Implantable medical devices must undergo thorough evaluation to ensure safety and efficacy before use in humans. If a device is designed to degrade, it is critical to understand the rate of degradation and the degradation products that will be released. Oxidative degradation is typically modeled in vitro by immersing materials or devices in hydrogen peroxide, which can limit further analysis of degradation products in many cases. Here we demonstrate a novel approach for testing the cytocompatibility of degradation products for oxidatively-degradable biomaterials where the materials are exposed to hydrogen peroxide, and then catalase enzyme is used to convert the hydrogen peroxide to water and oxygen so that the resulting aqueous solution can be added to cell culture media. To validate our results, expected degradation products are also synthesized then added to cell culture media. We used these methods to evaluate the cytocompatibility of degradation products from an oxidatively-degradable shape memory polyurethane designed in our lab and found that the degradation of these polymers is unlikely to cause a cytotoxic response in vivo based on the guidance provided by ISO 10993-5. These methods may also be applicable to other biocompatibility tests such as tests for mutagenicity or systemic toxicity, and evaluations of cell proliferation, migration, or gene and protein expression.

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