Modified DMEM xenic culture medium for propagation, isolation and maintenance of Balantioides coli

Yan, Wenchao; Wang, Tianqi; Zhao, Lizhuo; Sun, Chenyang

doi:10.1016/j.actatropica.2020.105762

Cited by 4 publications

(13 citation statements)

References 18 publications

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“…For performing in vitro cytotoxicity studies, a hepatocellular carcinoma cell line (HepG2) ( 38 , 39 ) was used. In the presence of a 5% CO 2 atmosphere, 100 mg/mL streptomycin, and 100 units/mL penicillin, the cells were cultured in a DMEM medium in the presence of a 10% phosphate buffer solution ( 40 ). The resultant cells were seeded in 96-well plates in the presence of 100 μL culture medium and kept for 24 hr.…”

Section: Methodsmentioning

confidence: 99%

Unveiling the therapeutic potential of cabozantinib-loaded poly D,L-lactic-co-glycolic acid and polysarcosine nanoparticles in inducing apoptosis and cytotoxicity in human HepG2 hepatocellular carcinoma cell lines and in vivo anti-tumor activity in SCID female mice

Bhattacharya

Parihar²,

Prajapati³

2023

Front. Oncol.

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IntroductionThe study aimed to develop a nano-based drug delivery system for the treatment of hepatocellular carcinoma (HCC), a type of liver cancer that accounts for 90% of all liver malignancies. The study focused on the use of cabozantinib (CNB), a potent multikinase inhibitor that targets the VEGF receptor 2, as the chemotherapeutic drug. We developed CNB-loaded nanoparticles made from Poly D, L-lactic-co-glycolic acid, and Polysarcosine (CNB-PLGA-PSar-NPs) for use in human HepG2 cell lines.MethodsBy O/W solvent evaporation method, the polymeric nanoparticles were prepared. The various techniques, such as photon correlation spectroscopy, scanning electron microscopy, and transmission electron microscopy were used, to determine the formulation's particle size, zeta potential, and morphology. SYBR Green/ROX qPCR Master Mix and RT-PCR equipment used to measure liver cancer cell line and tissue mRNA expression and MTT assay to test HepG2 cell cytotoxicity. Cell cycle arrest analysis, annexin V assay, and ZE5 Cell Analyzer apoptosis assay were also performed.ResultsThe results of the study showed that the particle diameters were 192.0 ± 3.67 nm with 0.128 PDI and -24.18 ± 3.34 mV zeta potential. The antiproliferative and proapoptotic effects of CNB-PLGA-PSar-NPs were evaluated using MTT and flow cytometry (FCM). The IC50 value of CNB-PLGA-PSar-NPs was 45.67 µg/mL, 34.73 µg/mL, and 21.56 µg/mL for 24, 48, and 72 h, respectively. The study also found that 11.20% and 36.77% of CNB-PLGA-PSar-NPs-treated cells were apoptotic at 60 µg/mL and 80 µg/mL, respectively, suggesting that the nanoparticles were effective in inducing apoptosis in the cancer cells. It can also conclude that, CNB-PLGA-PSar-NPs inhibit human HepG2 hepatocellular carcinoma cells and kill them by upregulating the tumour suppressor genes MT1F, MT1X, and downregulating MTTP, APOA4. Further in vivo antitumor activity was well reported in SCID female mice.DiscussionOverall, this study suggests that the CNB-PLGA-PSar-NPs are a promising drug delivery system for the treatment of HCC, and further research is needed to investigate their potential in clinical treatment.

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Section: Methodsmentioning

confidence: 99%

Unveiling the therapeutic potential of cabozantinib-loaded poly D,L-lactic-co-glycolic acid and polysarcosine nanoparticles in inducing apoptosis and cytotoxicity in human HepG2 hepatocellular carcinoma cell lines and in vivo anti-tumor activity in SCID female mice

Bhattacharya

Parihar²,

Prajapati³

2023

Front. Oncol.

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“…In the case of free-living ciliates, they can be obtained by inducing the excystment of the cysts; however, in the case of parasitic ciliates, excystment and maintenance of trophozoites in culture is complicated and often unfeasible. Protocols are available for in vitro culture of B. coli ( da Silva Barbosa et al, 2018a ; Yan et al, 2021 ), but they have some drawbacks (equipment requirements, culture conditions, low success rate) and cannot be used for routine diagnosis of the human ciliate . For a correct human illness source attribution ( Pires et al, 2009 ), molecular tools are the best option for a correct and accurate diagnosis of B. coli in both faecal and environmental samples.…”

Section: Diagnosismentioning

confidence: 99%

“…Data from in vitro cultures indicated that low pH values caused trophozoites to die ( da Silva Barbosa et al, 2015 ; Yan et al, 2021 ), so they would be unable to survive the low pH of the stomach ( Chalmers, 2014 ; Schuster and Visvesvara, 2004 ). Although Pomajbíková et al (2010) suggested that trophozoites might be infective, it is generally accepted that cysts are the infective stage of the parasite.…”

Section: Parasite Detection In Environmental Samplesmentioning

confidence: 99%

“…An alternative would be the use of in vitro models for the excystation and culture of B. coli , but more research is needed. Several media have been used for the culture of the trophozoites, with variable success rates ( Clark and Diamond, 2002 ; da Silva Barbosa et al, 2015 , da Silva Barbosa et al, 2018a ; Yan et al, 2021 ). In some cases, cultures were initiated from cysts ( da Silva Barbosa et al, 2015 ; Yan et al, 2021 ).…”

Section: Viability Assessment and Parasite Inactivationmentioning

confidence: 99%

“…Several media have been used for the culture of the trophozoites, with variable success rates ( Clark and Diamond, 2002 ; da Silva Barbosa et al, 2015 , da Silva Barbosa et al, 2018a ; Yan et al, 2021 ). In some cases, cultures were initiated from cysts ( da Silva Barbosa et al, 2015 ; Yan et al, 2021 ). However, the specific conditions for inducing the excystation process are undefined and da Silva Barbosa et al (2015) failed to obtain excystation in 36% of the cultures.…”

Section: Viability Assessment and Parasite Inactivationmentioning

confidence: 99%

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Cyst detection and viability assessment of Balantioides coli in environmental samples: Current status and future needs

García-Rodríguez

Köster

Ponce-Gordo

2022

Food and Waterborne Parasitology

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The ciliate Balantioides coli is a human enteric parasite that can cause life-threatening infections. It is a food- and waterborne parasite, with cysts being the infective stage. Despite its importance as a potential pathogen, few reports have investigated its presence in environmental samples, and some issues need attention including i) The accuracy of B. coli identification. In most cases, the protozoa is identified only by its morphological traits, which can be identical to those from other parasitic ciliates of animals. Genetic analysis of cysts recovered from environmental samples is necessary for species confirmation. In addition, genetic methods used with faecal samples need to be adequately validated with environmental matrices. ii) The methodology for searching this parasite in environmental samples. The protocols include an initial phase to isolate the cysts from the matrix followed by a second phase in which concentration procedures are usually applied. The methods may be valid but are not standardised and differences between studies could affect the results obtained. iii) The areas that needs further research. The development of genetic identification methods and standardised analytical protocols in environmental samples are required, as well as the assessment of viability and infectivity of B. coli cysts. The development of axenic culture systems will boost research on this parasite.

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Comparative Genomic and Transcriptomic Profiling Revealed the Molecular Basis of Starch Promoting the Growth and Proliferation of Balantioides coli

Zhao

Jiang

et al. 2023

Animals

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Carbohydrates are the main source of nutrition for B. coli, supplying energy for cell growth and development. The research aimed at investigating the mechanism of starch on the growth and replication of B. coli. Single-cell separation was used to isolate single trophozoites of B. coli under a stereomicroscope, transcriptomic profiling was conducted based on the SMART-seq2 single-cell RNA-seq method. Comparative genomic analysis was performed on B. coli and eight other ciliates to obtain specific and expanded gene families of B. coli. GO and KEGG enrichment analysis were used to analyze the key genes of B. coli under the action of starch in the present study. The results of single-cell RNA-seq depicts starch affected the growth and replication of B. coli in two ways: (1) the cell cycle was positively promoted by the activation of the cAMP/PKA signaling pathway via glycolysis; (2) the cell autophagy was suppressed through the PI3K/AKT/mTOR pathway. Genes involved in endocytosis, carbohydrate utilization, and the cAMP/PKA signaling pathway were highly enriched in both specific and expanded gene families of B. coli. Starch can be ingested and hydrolyzed into glucose, in turn affecting various biological processes of B. coli. The molecular mechanism of the effect of starch on the growth and proliferation of B. coli by promoting cell cycle and inhibiting the autophagy of trophozoites has been elucidated in our study.

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Modified DMEM xenic culture medium for propagation, isolation and maintenance of Balantioides coli

Cited by 4 publications

References 18 publications

Unveiling the therapeutic potential of cabozantinib-loaded poly D,L-lactic-co-glycolic acid and polysarcosine nanoparticles in inducing apoptosis and cytotoxicity in human HepG2 hepatocellular carcinoma cell lines and in vivo anti-tumor activity in SCID female mice

Unveiling the therapeutic potential of cabozantinib-loaded poly D,L-lactic-co-glycolic acid and polysarcosine nanoparticles in inducing apoptosis and cytotoxicity in human HepG2 hepatocellular carcinoma cell lines and in vivo anti-tumor activity in SCID female mice

Cyst detection and viability assessment of Balantioides coli in environmental samples: Current status and future needs

Comparative Genomic and Transcriptomic Profiling Revealed the Molecular Basis of Starch Promoting the Growth and Proliferation of Balantioides coli

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