Erratum to : “In Vitro application of the comet assay for aquatic genotoxicity: considering a primary culture versus a cell line” [Toxicology in Vitro 16 (2002) 177–184]

Abstract Aseptic loosening following periprosthetic osteolysis is the primary complication that limits the lifetime of total joint arthroplasty (TJA). The wear particles trigger a chronic inflammation response in the periprosthetic tissue and turn over the bone balance to bone resorption. The present study aimed to investigate the possible effect and mechanism of strontium ranelate (SR), a clinically safe drug for osteoporosis, on particle-induced periprosthetic osteolysis. Thirty-six female C57BL/6j mice underwent tibial Ti-nail implantation to establish an animal model of aseptic loosening. After 12 weeks, micro-CT results showed that strontium ranelate could inhibit periprosthetic bone resorption. In vitro, Ti particles were used to stimulate RAW264.7 cell line to collect conditioned medium, and co-culture MC3T3-E1 cell line with conditioned medium to establish a cell model of aseptic loosening. The results of alkaline phosphatase (ALP) detection, immunofluorescence, and flow cytometry demonstrated that strontium ranelate could regulate the expression of OPG/RANKL, promote differentiation and mineralization, and inhibit apoptosis in osteoblasts. Moreover, we revealed that SR’s exerted its therapeutic effect by down-regulating sclerostin, thereby activating the Wnt/β-catenin signal pathway. Therefore, this research suggests that strontium ranelate could be a potential drug for the prevention and treatment of particle-induced aseptic loosening post-TJA.

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Alkaline Comet Assay using the monocytic cell line THP-1

10.21203/rs.2.11936/v1 ◽

2019 ◽

Author(s):

Ana Neves-Costa ◽

Dora Pedroso ◽

Luis F Moita

Keyword(s):

Comet Assay ◽

Cell Line ◽

Adherent Cell ◽

Monocytic Cell ◽

Monocytic Cell Line ◽

Strand Breaks ◽

Human Monocytic Cell Line ◽

Human Monocytic Cell ◽

Dna Strand

Abstract This protocol details the experimental procedure for performing the comet assay, a very sensitive DNA break assay based on single cell gel electrophoresis.The analysis of DNA strand breaks, both single- and double-strand breaks (SSBs and DSBs, respectively), was performed in immune responsive cells. The cell line used was the human monocytic cell line THP-1, an adherent cell type with many known applications in in vitro studies of innate immunity. The comet assay is a robust procedure that allows the accurate and reproducible quantification of DNA damage. Here we describe not only the comet assay step-by-step protocol, but also some important aspects related to troubleshooting.

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Motility patterns of Trypanosoma cruzi trypomastigotes correlate with the efficiency of parasite invasion in vitro

Scientific Reports ◽

10.1038/s41598-020-72604-4 ◽

2020 ◽

Vol 10 (1) ◽

Cited By ~ 1

Author(s):

Jorge A. Arias-del-Angel ◽

Jesús Santana-Solano ◽

Moisés Santillán ◽

Rebeca G. Manning-Cela

Keyword(s):

Trypanosoma Cruzi ◽

Cell Line ◽

Mammalian Cells ◽

Dependent Manner ◽

Parasite Invasion ◽

Mammalian Cell Cultures ◽

A Cell ◽

Parasite Replication ◽

Parasite Motility

Abstract Numerous works have demonstrated that trypanosomatid motility is relevant for parasite replication and sensitivity. Nonetheless, although some findings indirectly suggest that motility also plays an important role during infection, this has not been extensively investigated. This work is aimed at partially filling this void for the case of Trypanosoma cruzi. After recording swimming T. cruzi trypomastigotes (CL Brener strain) and recovering their individual trajectories, we statistically analyzed parasite motility patterns. We did this with parasites that swim alone or above monolayer cultures of different cell lines. Our results indicate that T. cruzi trypomastigotes change their motility patterns when they are in the presence of mammalian cells, in a cell-line dependent manner. We further performed infection experiments in which each of the mammalian cell cultures were incubated for 2 h together with trypomastigotes, and measured the corresponding invasion efficiency. Not only this parameter varied from cell line to cell line, but it resulted to be positively correlated with the corresponding intensity of the motility pattern changes. Together, these results suggest that T. cruzi trypomastigotes are capable of sensing the presence of mammalian cells and of changing their motility patterns accordingly, and that this might increase their invasion efficiency.

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Establishment of a cell line of human endometrial adenocarcinoma in vitro

American Journal of Obstetrics and Gynecology ◽

10.1016/0002-9378(72)90861-7 ◽

1972 ◽

Vol 114 (8) ◽

pp. 1012-1019 ◽

Cited By ~ 162

Author(s):

Hiroyuki Kuramoto ◽

Shozo Tamura ◽

Yukio Notake

Keyword(s):

Cell Line ◽

Endometrial Adenocarcinoma ◽

A Cell

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Establishment of a cell line of human skin squamous cell carcinoma in vitro

British Journal of Dermatology ◽

10.1111/j.1365-2133.1981.tb01196.x ◽

1981 ◽

Vol 105 (2) ◽

pp. 125-132 ◽

Cited By ~ 47

Author(s):

S. KONDO ◽

K. ASO

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Cell Line ◽

Human Skin ◽

Squamous Cell ◽

Skin Squamous Cell Carcinoma ◽

A Cell

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Cytotoxicity of a cell-reactive immunotoxin containing ricin A chain is potentiated by an anti-immunotoxin containing ricin B chain.

Journal of Experimental Medicine ◽

10.1084/jem.160.1.341 ◽

1984 ◽

Vol 160 (1) ◽

pp. 341-346 ◽

Cited By ~ 46

Author(s):

E S Vitetta ◽

R J Fulton ◽

J W Uhr

Keyword(s):

Cell Surface ◽

Cell Line ◽

Ricin A Chain ◽

Daudi Cell ◽

A Cell ◽

A Chain ◽

Ricin A

In vitro killing of the human Daudi cell line by either univalent [F(ab')] or divalent (IgG) forms of rabbit anti-human Ig (RAHIg) coupled to ricin A chain can be specifically potentiated by a "piggyback" treatment with ricin B chain coupled to goat anti-rabbit Ig (GARIg). When cells are treated with univalent immunotoxin (IT) [F(ab') RAHIg-A] and then cultured, IT can be detected on the cell surface for at least 5 h, since GARIg-B can still enhance killing at this time. These results provide a strategy for in vivo use of A chain- and B chain-containing IT.

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Heterocyclic Compounds That Inhibit Rev-RRE Function and Human Immunodeficiency Virus Type 1 Replication

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00274-08 ◽

2008 ◽

Vol 52 (9) ◽

pp. 3169-3179 ◽

Cited By ~ 36

Author(s):

Deidra Shuck-Lee ◽

Fei Fei Chen ◽

Ryan Willard ◽

Sharmila Raman ◽

Roger Ptak ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Life Cycle ◽

Cell Line ◽

Particle Production ◽

Screening Assay ◽

Rev Response Element ◽

Immunodeficiency Virus ◽

A Cell ◽

Viruslike Particles

ABSTRACT A cell-based screening assay was performed to identify compounds that inhibited the postintegration stage of the human immunodeficiency virus (HIV) life cycle. This assay utilized a cell line that contains the HIV gag and pol genes expressed in a Rev-dependent fashion. The cell line produces about 10 to 15 ng of p24 per milliliter of medium over a 24-h period in the form of viruslike particles. Any compound that inhibits a postintegration step in the HIV life cycle scores in this assay by decreasing particle production. Forty thousand compounds were screened, and 192 compounds were selected from the original screen because they showed more than 50% inhibition at a 10 μM concentration. The cumulative evidence presented in this study strongly suggests that 2 of the 192 compounds work as inhibitors of HIV Rev function. This was determined by a variety of cell-based assays, although the compounds do not interfere with Rev-RRE (Rev response element) binding in vitro. Both compounds inhibit replication of the lab isolate NL4-3 as well as an HIV primary isolate from Brazil (93BR021) and thus are promising leads as therapeutic candidates that target HIV replication through inhibition of Rev function.

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