scholarly journals Proteomics Characterization of the Secretome from Rat Pancreatic Stellate Cells with ATP-Binding Cassette Transporters (ABCG2) and NCAM Phenotype

2013 ◽  
Vol 2013 ◽  
pp. 1-18
Author(s):  
Maria Lucas ◽  
Eugenia Mato ◽  
Silvia Barceló-Batllori ◽  
Ramon Gomis ◽  
Anna Novials
Keyword(s):  
Conditioned Medium ◽  
Cell Population ◽  
Stellate Cells ◽  
Resistant Cell ◽  
Pancreatic Stellate Cells ◽  
Cellular Processes ◽  
Proliferation And Differentiation ◽  
Development And Differentiation ◽  
Differentiation Pathways

We have previously reported the identification of a pancreata mitoxantrone-resistant cell population which expressed the ABCG2 transporter with a pancreatic stellate cells phenotype (PaSC) and ability of secreting insulin after inducing their differentiation. The characterization of the secretome of this cell population by two-dimensional electrophoresis (2D) coupled with mass spectrometry MALDI-TOF was able to identify seventy-six protein spots involved in different cellular processes: development/differentiation, proteases, immune response, and other. Moreover, Ingenuity Pathway Analysis displayed several significant networks and TGFβ1 molecule was identified as a central node of one of them. The effect of this active molecule secreted in the conditioned medium was investigated in ductal cell line (ARIP). The results showed that the conditioned medium inhibited their proliferation without affecting their cell viability. Additionally, they showed an upregulation of PDX1 and downregulation of CK19. The rate of ARIP cell proliferation was recovered, but no effects on the gene expression were observed after using TGFβ1-neutralising antibody. Proteins associated with cell growth, development and differentiation such as PEDF, LIF, and Wnt5b, identified in the secretome, could be involved in the observed transcription changes. These finding may suggest a new paracrine action of PaSCs involved in the proliferation and differentiation pathways not yet identified.

scholarly journals About the dangers, costs and benefits of living an aerobic lifestyle

2014 ◽  
Vol 42 (4) ◽  
pp. 917-921 ◽  
Author(s):  
Daniela Knoefler ◽  
Lars I.O. Leichert ◽  
Maike Thamsen ◽  
Claudia M. Cremers ◽  
Dana Reichmann ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Research Programme ◽  
Future Research ◽  
Mammalian Host ◽  
Costs And Benefits ◽  
Cellular Processes ◽  
Effective Antioxidant ◽  
Development And Differentiation ◽  
Micro Organisms

The era in which ROS (reactive oxygen species) were simply the ‘bad boys of biology’ is clearly over. High levels of ROS are still rightfully considered to be toxic to many cellular processes and, as such, contribute to disease conditions and cell death. However, the high toxicity of ROS is also extremely beneficial, particularly as it is used to kill invading micro-organisms during mammalian host defence. Moreover, a transient, often more localized, increase in ROS levels appears to play a major role in signal transduction processes and positively affects cell growth, development and differentiation. At the heart of all these processes are redox-regulated proteins, which use oxidation-sensitive cysteine residues to control their function and by extension the function of the pathways that they are part of. Our work has contributed to changing the view about ROS through: (i) our characterization of Hsp33 (heat-shock protein 33), one of the first redox-regulated proteins identified, whose function is specifically activated by ROS, (ii) the development of quantitative tools that reveal extensive redox-sensitive processes in bacteria and eukaryotes, and (iii) the discovery of a link between early exposure to oxidants and aging. Our future research programme aims to generate an integrated and system-wide view of the beneficial and deleterious effects of ROS with the central goal to develop more effective antioxidant strategies and more powerful antimicrobial agents.

scholarly journals Role of c-MET Inhibitors in Overcoming Drug Resistance in Spheroid Models of Primary Human Pancreatic Cancer and Stellate Cells

Cancers ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 638 ◽  
Author(s):  
Omidreza Firuzi ◽  
Pei Pei Che ◽  
Btissame El Hassouni ◽  
Mark Buijs ◽  
Stefano Coppola ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Conditioned Medium ◽  
Therapeutic Option ◽  
Stellate Cells ◽  
Luciferase Assay ◽  
Pancreatic Stellate Cells ◽  
Human Pancreatic Cancer ◽  
Ductal Adenocarcinoma ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fluorescence Protein

Pancreatic stellate cells (PSCs) are a key component of tumor microenvironment in pancreatic ductal adenocarcinoma (PDAC) and contribute to drug resistance. c-MET receptor tyrosine kinase activation plays an important role in tumorigenesis in different cancers including PDAC. In this study, effects of PSC conditioned medium (PCM) on c-MET phosphorylation (by immunocytochemistry enzyme-linked immunosorbent assay (ELISA)) and drug response (by sulforhodamine B assay) were investigated in five primary PDAC cells. In novel 3D-spheroid co-cultures of cyan fluorescence protein (CFP)-firefly luciferase (Fluc)-expressing primary human PDAC cells and green fluorescence protein (GFP)-expressing immortalized PSCs, PDAC cell growth and chemosensitivity were examined by luciferase assay, while spheroids’ architecture was evaluated by confocal microscopy. The highest phospho-c-MET expression was detected in PDAC5 and its subclone sorted for “stage specific embryonic antigen-4” (PDAC5 (SSEA4)). PCM of cells pre-incubated with PDAC conditioned medium, containing increased hepatocyte growth factor (HGF) levels, made PDAC cells significantly more resistant to gemcitabine, but not to c-MET inhibitors. Hetero-spheroids containing both PSCs and PDAC5 (SSEA4) cells were more resistant to gemcitabine compared to PDAC5 (SSEA4) homo-spheroids. However, c-MET inhibitors (tivantinib, PHA-665752 and crizotinib) were equally effective in both spheroid models. Experiments with primary human PSCs confirmed the main findings. In conclusion, we developed spheroid models to evaluate PSC–PDAC reciprocal interaction, unraveling c-MET inhibition as an important therapeutic option against drug resistant PDAC.

Identification, culture, and characterization of pancreatic stellate cells in rats and humans

1998 ◽  
Vol 115 (2) ◽  
pp. 421-432 ◽  
Author(s):  
Max G. Bachem ◽  
Erik Schneider ◽  
Hans Groß ◽  
Hans Weidenbach ◽  
Roland M. Schmid ◽  
...  
Keyword(s):  
Stellate Cells ◽  
Pancreatic Stellate Cells

In vitro characterization of different primary and immortalized pancreatic stellate cells

Pancreatology ◽  
2018 ◽  
Vol 18 (4) ◽  
pp. S151
Author(s):  
Daniela Lenggenhager ◽  
Manoj Amrutkar ◽  
Petra Sántha ◽  
Monica Aasrum ◽  
Matthias Löhr ◽  
...  
Keyword(s):  
Stellate Cells ◽  
Pancreatic Stellate Cells ◽  
In Vitro Characterization

Characterization of Tumor-Derived Pancreatic Stellate Cells

2009 ◽  
Vol 157 (1) ◽  
pp. 96-102 ◽  
Author(s):  
Buckminster Farrow ◽  
David Rowley ◽  
Truong Dang ◽  
David H. Berger
Keyword(s):  
Stellate Cells ◽  
Pancreatic Stellate Cells

Identification of a pancreatic stellate cell population with properties of progenitor cells: new role for stellate cells in the pancreas

2009 ◽  
Vol 421 (2) ◽  
pp. 181-191 ◽  
Author(s):  
Eugenia Mato ◽  
Maria Lucas ◽  
Jordi Petriz ◽  
Ramon Gomis ◽  
Anna Novials
Keyword(s):  
Progenitor Cells ◽  
Cell Population ◽  
Glucose Transporter ◽  
Stellate Cell ◽  
Stellate Cells ◽  
Pancreatic Stellate Cells ◽  
Diabetes Research ◽  
Pancreatic Cell ◽  
Cell Clusters ◽  
Ultrastructural Studies

Numerous studies conducted in a diversity of adult tissues have shown that certain stem cells are characterized by the expression of a protein known as the ABCG2 transporter (where ABC is ATP- binding cassette). In the adult pancreas, although various multipotent progenitors have been proposed, the ABCG2 marker has only been detected in the so-called ‘side population’ (a primitive haematopoietic cell population with a multipotential capacity). In the present study we sought to identify new ABCG2+ pancreatic cell populations and to explore whether they exhibit the properties of progenitor cells. We isolated and expanded mitoxantrone-resistant cells from pancreata of lactating rats by drug selection. These cells were characterized and maintained in different stages of differentiation using several media ‘cocktails’ plus Matrigel™ (BD Biosciences). Differentiation was assessed by RT–PCR (reverse transcription–PCR), immunocytochemistry, electron microscopy and ELISA. The expanded cell population demonstrated a phenotype of PaSCs (pancreatic stellate cells). Spontaneous cell clusters occurred during cell expansion and they showed weak expression of the transcription factor Pdx1 (pancreatic and duodenal homeobox 1). Moreover, the presence of inductive factors in the Matrigel plus exendin-4 led to an increase in Pdx1 and endocrine genes, such as insulin, islet amyloid polypeptide, glucagon, the glucose transporter GLUT2, chromogranin A and the convertases PC1/3 and PC2 were also detected. Immunocytochemical analysis showed co-localization of insulin and C-peptide, whereas ultrastructural studies revealed the presence of granules. Insulin secretion from cell clusters was detected in the cell culture medium. We identified a population of PaSCs that express the ABCG2+ transporter and have the capacity to transdifferentiate into insulin-producing cells. Although the potential therapeutic application remains to be tested, PaSCs could represent a future option for insulin replacement in diabetes research.

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