In Vitro Enzymatic and Cell Culture-Based Assays for Measuring Activity of HBV RNaseH Inhibitors

ABSTRACT There are several in vitro methods to analyse the function of the adenohypophysis or the mechanisms of its regulation. The present paper deals with single cell culture, organ culture and short term incubation techniques by which the morphology and gonadotrophin-secreting function of the adenohypophysis were studied. In trypsin-dispersed cell culture, the adenohypophysial cells showed extensive propagation to form numerous cell colonies and finally develop into a confluent monolayer cell sheet covering completely the surface of culture vessels. Almost all of the cultured cells, however, became chromophobic, at least at the end of the first week of cultivation, when gonadotrophin was detectable neither in the culture medium nor in the cells themselves. After the addition of the hypothalamic extract, gonadotrophin became detectable again, and basophilic or PAS-positive granules also reappeared within the cells, suggesting that the gonadotrophs were stimulated by the extract to produce gonadotrophin. In organ culture and short term incubation, the incorporation of [3H] leucine into the adenohypophysial cells in relation to the addition of hypothalamic extract was examined. It was obvious that the ability to incorporate [3H] leucine into the gonadotrophs in vitro was highly dependent upon the presence of the hypothalamic extract.

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Induction of melanocyte precursors from neural crest cells surrounding the neural tube-like structures developed in vitro using mouse ES cell culture

Inflammation and Regeneration ◽

10.2492/inflammregen.27.45 ◽

2007 ◽

Vol 27 (1) ◽

pp. 45-52

Author(s):

Koh-ichi Atoh ◽

Manae S. Kurokawa ◽

Hideshi Yoshikawa ◽

Chieko Masuda ◽

Erika Takada ◽

...

Keyword(s):

Cell Culture ◽

Neural Crest ◽

Neural Tube ◽

Neural Crest Cells ◽

Es Cell ◽

Mouse Es Cell

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Evaluation of Diallyl Sulfide Analogs for Cytotoxicity, Inhibition of CYP2E1, and Metabolite Profiling Using In Vitro Cell Culture, Human Liver Microsomes and LCMS/MS

10.26226/morressier.57d6b2b8d462b8028d88ee12 ◽

2016 ◽

Author(s):

Mohammad Rahman

Keyword(s):

Cell Culture ◽

Metabolite Profiling ◽

Human Liver ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Diallyl Sulfide ◽

In Vitro Cell Culture

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Comparison of pCR Rate After Neadjuvant Chemotherapy Guided by Result of in Vitro Cell Culture Drug Sensitivity or Physician's Choice

Case Medical Research ◽

10.31525/ct1-nct04130750 ◽

2019 ◽

Author(s):

Keyword(s):

Cell Culture ◽

Drug Sensitivity ◽

In Vitro Cell Culture

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Obtaining and Characterization of Alhagi persarum Boiss. et Buhse Callus Cell Cultures, Isoflavonoid Producers

Biotekhnologiya ◽

10.21519/0234-2758-2020-36-6-35-48 ◽

2020 ◽

Vol 36 (6) ◽

pp. 35-48

Author(s):

D.V. Коchkin ◽

G.I. Sobolkovа ◽

А.А. Fоmеnkov ◽

R.А. Sidorov ◽

А.М. Nоsоv

Keyword(s):

Fatty Acids ◽

Cell Culture ◽

Liquid Chromatography ◽

Secondary Metabolites ◽

Cell Cultures ◽

Mass Spectrometric ◽

Mass Spectrometric Detection ◽

Gas Liquid Chromatography ◽

Callus Cell

The physiological characteristics of the callus cell cultures of Alhagi persarum Boiss et Buhse, a member of the legume family, widely used in folk medicine, have been studied. It was shown that the source of the explant was an important factor in the initiation of callusogenesis: more intense callusogenesis (almost 100%) was observed for explants from various organs of sterile seedlings, rather than intact plants (less than 30%). As a result, more than 20 lines of morphologically different callus cell cultures were obtained, and the growth parameters for the 5 most intensively growing lines were determined. The composition of fatty acids (FA) of total lipids and secondary metabolites in the most physiologically stable callus line Aр-207 was analyzed. Using capillary gas-liquid chromatography with mass spectrometric detection (GLC-MS), 19 individual C12--C24 FAs were identified, the main fraction of which were palmitic (~ 23%), stearic (~ 22%), linoleic (~ 14%) and α-linolenic (~ 33%) acids. The established atypical ratio of FAs (a simultaneous high content of both saturated FAs and polyunsaturated α-linolenic acid) is possibly due to the adaptation of cells to in vitro growth conditions. Phytochemical analysis of the secondary metabolites was carried out using ultra-performance liquid chromatography with electrospray ionization mass spectrometric detection (UPLC MS). Compounds belonging to different structural groups of isoflavones were found. Aglycones (calycosin, formononetin and afrormosin isomer), glucosides (formononetin glucoside), as well as esters of glucosides (malonylglycosides of calicosin, formononetin, afrormosin isomers, glycitein and genistein) were detected. These secondary metabolites are widespread in plants of the Fabaceae family; however, isoflavones are rare in representatives of the Alhagi genus. The presence of malonylated isoflavone glycosides in Alhagi spp. was shown for the first time. endemic plant species, Alhagi, in vitro cell culture, callus cell culture, isoflavones, fatty acids All studies were carried out using the equipment of the "Experimental Biotechnological Facility" and the "All-Russian Collection of Cell Cultures of Higher Plants" of IРР RAS. This work was supported by the Russian Foundation for Basic Research (RFBR), contract no.18-54-06021 (Az_a), and the Government of the Russian Federation, Megagrant Project no. 075-15-2019-1882.

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Recent Progress in Prediction Systems for Drug-induced Liver Injury Using in vitro Cell Culture

Drug Metabolism Letters ◽

10.2174/1872312814666201202112610 ◽

2020 ◽

Vol 14 ◽

Author(s):

Shogo Ozawa ◽

Toshitaka Miura ◽

Jun Terashima ◽

Wataru Habano ◽

Seiichi Ishida

Keyword(s):

Cell Culture ◽

Recent Progress ◽

Inflammatory Cells ◽

Protein Adducts ◽

In Vitro Assays ◽

Drug Induced ◽

Drug Induced Liver Injury ◽

Culture Systems ◽

Cell Culture Systems

Background: In order to avoid drug-induced liver injury (DILI), in vitro assays, which enable the assessment of both metabolic activation and immune reaction processes that ultimately result in DILI, are needed. Objective: In this study, the recent progress in the application of in vitro assays using cell culture systems is reviewed for potential DILI-causing drugs/xenobiotics and a mechanistic study on DILI, as well as for the limitations of in vitro cell culture systems for DILI research. Methods: Information related to DILI was collected through a literature search of the PubMed database. Results: The initial biological event for the onset of DILI is the formation of cellular protein adducts after drugs have been metabolically activated by drug metabolizing enzymes. The damaged peptides derived from protein adducts lead to the activation of CD4+ helper T lymphocytes and recognition by CD8+ cytotoxic T lymphocytes, which destroy hepatocytes through immunological reactions. Because DILI is a major cause of drug attrition and drug withdrawal, numerous in vitro systems consisting of hepatocytes and immune/inflammatory cells, or spheroids of human primary hepatocytes containing non-parenchymal cells have been developed. These cellular-based systems have identified DILIinducing drugs with approximately 50% sensitivity and 90% specificity. Conclusion: Different co-culture systems consisting of human hepatocyte-derived cells and other immune/inflammatory cells have enabled the identification of DILI-causing drugs and of the actual mechanisms of action.

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The URNA Genes of Herpesvirus Saimiri (Strain C488) Are Dispensable for Transformation of Human T Cells In Vitro

Journal of Virology ◽

10.1128/jvi.73.12.10551-10555.1999 ◽

1999 ◽

Vol 73 (12) ◽

pp. 10551-10555 ◽

Cited By ~ 32

Author(s):

Armin Ensser ◽

André Pfinder ◽

Ingrid Müller-Fleckenstein ◽

Bernhard Fleckenstein

Keyword(s):

Cell Culture ◽

Herpesvirus Saimiri ◽

Wild Type Virus ◽

Type Virus ◽

Wild Type ◽

Human T Cell ◽

Human T Cells ◽

Small Nuclear Rnas ◽

T Cell Lines

ABSTRACT The herpesvirus saimiri strain C488 genome contains five genes for small nuclear RNAs, termed herpesvirus saimiri URNAs (or HSURs). Using a cosmid-based approach, all HSURs were precisely deleted from the genome. The mutant virus replicated at levels that were similar to those of wild-type viruses in OMK cells. Although the HSURs are expressed in wild-type virus-transformed human T-cell lines, the deletion does not affect viral transformation in cell culture.

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Effect of cat litters on feline coronavirus infection of cell culture and cats

Journal of Feline Medicine and Surgery ◽

10.1177/1098612x19848167 ◽

2019 ◽

Vol 22 (4) ◽

pp. 350-357 ◽

Cited By ~ 6

Author(s):

Diane Addie ◽

Lene Houe ◽

Kirsty Maitland ◽

Giuseppe Passantino ◽

Nicola Decaro

Keyword(s):

Cell Culture ◽

Virus Infection ◽

Real Life ◽

In Vitro Study ◽

Oral Route ◽

Virus Shedding ◽

Virus Load ◽

Fuller’S Earth ◽

Feline Coronavirus

Objectives Feline infectious peritonitis (FIP) is caused by infection with feline coronavirus (FCoV). FCoV is incredibly contagious and transmission is via the faecal–oral route. FCoV infection, and therefore FIP, is most common in breeder and rescue catteries, where many cats are kept indoors, using litter trays. Whether it is possible to break the cycle of FCoV infection and reinfection using cat litters has never been investigated. The aim of the study was to examine the effect of cat litters on FCoV infectivity and virus load in multi-cat households, and transmission frequency. Methods Fifteen cat litters were mixed and incubated with FCoV, centrifuged and the supernatants tested in vitro for the ability to prevent virus infection of cell culture. To test applicability of in vitro results to real life, virus load was measured in two households in a double crossover study of four Fuller’s earth-based cat litters by testing rectal swabs using FCoV reverse transcriptase quantitative PCR. Results Four litters abrogated FCoV infection of cell culture, nine reduced it to a greater or lesser extent and two had no effect. One brand had different virus inhibitory properties depending on where it was manufactured. Fuller’s earth-based litters performed best, presumably by adsorbing virus. In the field study, there appeared to be less virus shedding on one Fuller’s earth-based cat litter. Conclusions and relevance The in vitro study successfully identified cat litters that inactivate FCoV; such litters exist so do not need to be developed. Fuller’s earth-based litters best prevented infection of cell culture, but did not completely abrogate FCoV transmission in two multi-cat households. A dust-free clumping Fuller’s earth litter appeared to fare best, but virus shedding also varied on the control litters, complicating interpretation. Sawdust-based cat litters are not useful in FCoV-endemic households because they track badly and have a poor effect on virus infection.

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