scholarly journals AnIn VitroCulture System for Long-Term Expansion of Epithelial and Mesenchymal Salivary Gland Cells: Role of TGF-β1 in Salivary Gland Epithelial and Mesenchymal Differentiation

2013 ◽  
Vol 2013 ◽  
pp. 1-20 ◽  
Author(s):  
Kajohnkiart Janebodin ◽  
Worakanya Buranaphatthana ◽  
Nicholas Ieronimakis ◽  
Aislinn L. Hays ◽  
Morayma Reyes
Keyword(s):  
Salivary Gland ◽  
Culture System ◽  
Gland Cell ◽  
Mesenchymal Cells ◽  
Salivary Gland Cell ◽  
Salivary Gland Cells ◽  
Gland Development

Despite a pivotal role in salivary gland development, homeostasis, and disease, the role of salivary gland mesenchyme is not well understood. In this study, we used theCol1a1-GFPmouse model to characterize the salivary gland mesenchymein vitroandin vivo. TheCol1a1-GFPtransgene was exclusively expressed in the salivary gland mesenchyme.Ex vivoculture of mixed salivary gland cells in DMEM plus serum medium allowed long-term expansion of salivary gland epithelial and mesenchymal cells. The role of TGF-β1 in salivary gland development and disease is complex. Therefore, we used thisin vitroculture system to study the effects of TGF-β1 on salivary gland cell differentiation. TGF-β1 induced the expression of collagen, and inhibited the formation of acini-like structures in close proximity to mesenchymal cells, which adapted a fibroblastic phenotype. In contrast, TGF-βR1 inhibition increased acini genes and fibroblast growth factors (Fgf-7andFgf-10), decreased collagen and induced formation of larger, mature acini-like structures. Thus, inhibition of TGF-βsignaling may be beneficial for salivary gland differentiation; however, due to differential effects of TGF-β1 in salivary gland epithelial versus mesenchymal cells, selective inhibition is desirable. In conclusion, this mixed salivary gland cell culture system can be used to study epithelial-mesenchymal interactions and the effects of differentiating inducers and inhibitors.

scholarly journals Non-Lethal Concentration of MeHg Causes Marked Responses in the DNA Repair, Integrity, and Replication Pathways in the Exposed Human Salivary Gland Cell Line

2021 ◽  
Vol 12 ◽  
Author(s):  
Lygia Sega Nogueira ◽  
Carolina P. Vasconcelos ◽  
Jessica Rodrigues Plaça ◽  
Geovanni Pereira Mitre ◽  
Leonardo Oliveira Bittencourt ◽  
...  
Keyword(s):  
Dna Repair ◽  
Salivary Gland ◽  
Cell Line ◽  
Gland Cell ◽  
Salivary Gland Cell ◽  
Human Salivary Gland ◽  
Gland Cells ◽  
Salivary Gland Cells

In Brazilian northern Amazon, communities are potentially exposed and vulnerable to methylmercury (MeHg) toxicity through the vast ingestion of fish. In vivo and in vitro studies demonstrated that the salivary glands as a susceptible organ to this potent environmental pollutant, reporting alterations on physiological, biochemical, and proteomic parameters. However, the alterations caused by MeHg on the gene expression of the exposed human salivary gland cells are still unknown. Therefore, the goal was to perform the transcriptome profile of the human salivary gland cell line after exposure to MeHg, using the microarray technique and posterior bioinformatics analysis. The cell exposure was performed using 2.5 µM MeHg. A previously published study demonstrated that this concentration belongs to a range of concentrations that caused biochemical and metabolic alterations in this linage. As a result, the MeHg exposure did not cause lethality in the human salivary gland cells line but was able to alter the expression of 155 genes. Downregulated genes (15) are entirety relating to the cell metabolism impairment, and according to KEGG analysis, they belong to the glycosphingolipid (GSL) biosynthesis pathway. On the other hand, most of the 140 upregulated genes were related to cell-cycle progression, DNA repair, and replication pathway, or cellular defenses through the GSH basal metabolism. These genomic changes revealed the effort to the cell to maintain physiological and genomic stability to avoid cell death, being in accordance with the nonlethality in the toxicity test. Last, the results support in-depth studies on nonlethal MeHg concentrations for biomarkers identification that interpret transcriptomics data in toxicological tests serving as an early alert of physiological changes in vitro biological models.

scholarly journals Functional analysis of the biochemical activity of mammalian phosphatidylinositol 5 phosphate 4-kinase enzymes

2019 ◽  
Vol 39 (2) ◽  
Author(s):  
Swarna Mathre ◽  
K. Balasankara Reddy ◽  
Visvanathan Ramya ◽  
Harini Krishnan ◽  
Avishek Ghosh ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Cell Size ◽  
Kinase Activity ◽  
Gland Cell ◽  
Specific Activity ◽  
Salivary Gland Cell ◽  
Drosophila Genome ◽  
Biochemical Activity

Abstract Phosphatidylinositol 5 phosphate 4-kinase (PIP4K) are enzymes that catalyse the phosphorylation of phosphatidylinositol 5-phosphate (PI5P) to generate PI(4,5)P2. Mammalian genomes contain three genes, PIP4K2Α, 2B and 2C and murine knockouts for these suggested important physiological roles in vivo. The proteins encoded by PIP4K2A, 2B and 2C show widely varying specific activities in vitro; PIP4K2A is highly active and PIP4K2C 2000-times less active, and the relationship between this biochemical activity and in vivo function is unknown. By contrast, the Drosophila genome encodes a single PIP4K (dPIP4K) that shows high specific activity in vitro and loss of this enzyme results in reduced salivary gland cell size in vivo. We find that the kinase activity of dPIP4K is essential for normal salivary gland cell size in vivo. Despite their highly divergent specific activity, we find that all three mammalian PIP4K isoforms are able to enhance salivary gland cell size in the Drosophila PIP4K null mutant implying a lack of correlation between in vitro activity measurements and in vivo function. Further, the kinase activity of PIP4K2C, reported to be almost inactive in vitro, is required for in vivo function. Our findings suggest the existence of unidentified factors that regulate PIP4K enzyme activity in vivo.

scholarly journals Tumor cell–organized fibronectin maintenance of a dormant breast cancer population

2020 ◽  
Vol 6 (11) ◽  
pp. eaaz4157 ◽  
Author(s):  
Lauren E. Barney ◽  
Christopher L. Hall ◽  
Alyssa D. Schwartz ◽  
Akia N. Parks ◽  
Christopher Sparages ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Culture System ◽  
Cell Populations ◽  
Cell Culture System ◽  
Proliferating Cells ◽  
In Vitro Method ◽  
Fibronectin Deposition

Tumors can undergo long periods of dormancy, with cancer cells entering a largely quiescent, nonproliferative state before reactivation and outgrowth. To understand the role of the extracellular matrix (ECM) in regulating tumor dormancy, we created an in vitro cell culture system with carefully controlled ECM substrates to observe entrance into and exit from dormancy with live imaging. We saw that cell populations capable of surviving entrance into long-term dormancy were heterogeneous, containing quiescent, cell cycle–arrested, and actively proliferating cells. Cell populations capable of entering dormancy formed an organized, fibrillar fibronectin matrix via αvβ3 and α5β1 integrin adhesion, ROCK-generated tension, and TGFβ2 stimulation, and cancer cell outgrowth after dormancy required MMP-2–mediated fibronectin degradation. We propose this approach as a useful, in vitro method to study factors important in regulating dormancy, and we used it here to elucidate a role for fibronectin deposition and MMP activation.

scholarly journals Stochastic model of BKPy Virus replication and assembly

2019 ◽  
Author(s):  
Suzy M. Stiegelmeyer ◽  
Liesl K. Jeffers-Francis ◽  
Morgan C. Giddings ◽  
Jennifer Webster-Cyriaque
Keyword(s):  
Salivary Gland ◽  
Self Assembly ◽  
Gland Cell ◽  
Jc Virus ◽  
Viral Proteins ◽  
Salivary Gland Cell ◽  
Oral Transmission ◽  
Gland Cells ◽  
Complex Processes ◽  
Salivary Gland Cells

AbstractBK Polyomavirus (BKPyV), belongs to the same family as SV40 and JC Virus and has recently been associated with both Sjögrens Syndrome and HIV associated Salivary Gland Disease. BKPyV was previously only known for causing the rejection of kidney transplants. As such, BKPyV infection of salivary gland cells implicates oral transmission of the virus. BKPyV replicates slowly in salivary gland cells, producing infectious virus after 72-96 hours. However, it remains unclear how this virus infects or replicates within salivary gland cells, blocking the development of therapeutic strategies to inhibit the virus. Thus, an intracellular, computational model using agent-based modeling was developed to model BKPyV replication within a salivary gland cell. In addition to viral proteins, we modeled host cell machinery that aids transcription, translation and replication of BKPyV. The model has separate cytosolic and nuclear compartments, and represents all large molecules such as proteins, RNAs, and DNA as individual computer “agents” that move and interact within the simulated salivary gland cell environment. An application of the Boids algorithm was implemented to simulate molecular binding and formation of BKPyV virions and BKPyV virus-like particles (VLPs). This approach enables the direct study of spatially complex processes such as BKPyV virus self-assembly, transcription, and translation. This model reinforces experimental results implicating the processes that result in the slow accumulation of viral proteins. It revealed that the slow BKPyV replication rate in salivary gland cells might be explained by capsid subunit accumulation rates. BKPyV particles may only form after large concentrations of capsid subunits have accumulated. In addition, salivary gland specific transcription factors may enable early region transcription of BKPyV.

scholarly journals Perlecan Domain IV Peptide Stimulates Salivary Gland Cell Assembly In Vitro

2009 ◽  
Vol 15 (11) ◽  
pp. 3309-3320 ◽  
Author(s):  
Swati Pradhan ◽  
Chu Zhang ◽  
Xinqiao Jia ◽  
Daniel D. Carson ◽  
Robert Witt ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Gland Cell ◽  
Salivary Gland Cell ◽  
Cell Assembly

scholarly journals Novel Modeling Approach to Generate a Polymeric Nanofiber Scaffold for Salivary Gland Cells

2010 ◽  
Vol 1 (3) ◽  
Author(s):  
Riffard Jean-Gilles ◽  
David Soscia ◽  
Sharon Sequeira ◽  
Michael Melfi ◽  
Anand Gadre ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Gland Cell ◽  
Self Organization ◽  
Salivary Gland Cell ◽  
Nanofiber Scaffold ◽  
Salivary Gland Tissue ◽  
Salivary Gland Cells ◽  
Gland Tissue ◽  
Nanofiber Scaffolds ◽  
Tissue Construct

Electrospun nanofibers have been utilized in many biomedical applications as biomimetics of extracellular matrix proteins that promote self-organization of cells into 3D tissue constructs. As progress toward an artificial salivary gland tissue construct, we prepared nanofiber scaffolds using PLGA, which is a biodegradable and biocompatible material. We used electrospinning to prepare nanofiber scaffolds using poly(lactic-co-glycolic acid) (PLGA) with both dimethylformamide (DMF) and hexafluoroisopropanol (HFIP) as solvents. Using a design of experiment approach, the system and process parameters were optimized concurrently, and their effects on the diameter of the resulting fibers were computed into a single model. A transfer function was used to reproducibly produce nanofibers of a defined diameter, which was confirmed by a scanning electron microscope. The salivary gland cell line was seeded on the nanofiber scaffolds, and morphology, cell proliferation, and viability were assayed. Varying two or more parameters simultaneously yielded trends diverging from the linear response predicted by previous studies. Comparison of two solvents revealed that the diameter of PLGA nanofibers generated using HFIP is less sensitive to changes in the system and process parameters than are fibers generated using DMF. Inclusion of NaCl reduced morphological inconsistencies and minimized process variability. The resulting nanofiber scaffolds supported attachment, survival, and cell proliferation of a mouse salivary gland epithelial cell line. In comparison with glass and flat PLGA films, the nanofibers promoted self-organization of the salivary gland cells into 3D cell clusters, or aggregates. These data indicate that nanofiber scaffolds promote salivary gland cell organization, and suggest that a nanofiber scaffold could provide a platform for engineering of an artificial salivary gland tissue construct. This study additionally provides a method for efficient production of nanofiber scaffolds for general application in tissue engineering.

THE ROLE OF POLYETHYLENIMINE IN ENHANCING PERFORMANCE OF GLUTAMATE BIOSENSORS

2018 ◽  
Vol 8 (3) ◽  
pp. 36-41
Author(s):  
Diep Do Thi Hong ◽  
Duong Le Phuoc ◽  
Hoai Nguyen Thi ◽  
Serra Pier Andrea ◽  
Rocchitta Gaia
Keyword(s):  
First Generation ◽  
Good Reproducibility ◽  
Complex Matrices ◽  
Long Term Stability ◽  
In Vitro Characterization ◽  
Glutamate Oxidase

Background: The first biosensor was constructed more than fifty years ago. It was composed of the biorecognition element and transducer. The first-generation enzyme biosensors play important role in monitoring neurotransmitter and determine small quantities of substances in complex matrices of the samples Glutamate is important biochemicals involved in energetic metabolism and neurotransmission. Therefore, biosensors requires the development a new approach exhibiting high sensibility, good reproducibility and longterm stability. The first-generation enzyme biosensors play important role in monitoring neurotransmitter and determine small quantities of substances in complex matrices of the samples. The aims of this work: To find out which concentration of polyethylenimine (PEI) exhibiting the most high sensibility, good reproducibility and long-term stability. Methods: We designed and developed glutamate biosensor using different concentration of PEI ranging from 0% to 5% at Day 1 and Day 8. Results: After Glutamate biosensors in-vitro characterization, several PEI concentrations, ranging from 0.5% to 1% seem to be the best in terms of VMAX, the KM; while PEI content ranging from 0.5% to 1% resulted stable, PEI 1% displayed an excellent stability. Conclusions: In the result, PEI 1% perfomed high sensibility, good stability and blocking interference. Furthermore, we expect to develop and characterize an implantable biosensor capable of detecting glutamate, glucose in vivo. Key words: Glutamate biosensors, PEi (Polyethylenimine) enhances glutamate oxidase, glutamate oxidase biosensors

scholarly journals Chemically defined and xeno-free culture condition for human extended pluripotent stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Bei Liu ◽  
Shi Chen ◽  
Yaxing Xu ◽  
Yulin Lyu ◽  
Jinlin Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Culture Condition ◽  
Human Fibroblast ◽  
Culture System ◽  
Disease Modeling ◽  
Directed Differentiation

AbstractExtended pluripotent stem (EPS) cells have shown great applicative potentials in generating synthetic embryos, directed differentiation and disease modeling. However, the lack of a xeno-free culture condition has significantly limited their applications. Here, we report a chemically defined and xeno-free culture system for culturing and deriving human EPS cells in vitro. Xeno-free human EPS cells can be long-term and genetically stably maintained in vitro, as well as preserve their embryonic and extraembryonic developmental potentials. Furthermore, the xeno-free culturing system also permits efficient derivation of human EPS cells from human fibroblast through reprogramming. Our study could have broad utility in future applications of human EPS cells in biomedicine.

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