Biological Application of a Fluorescent Zinc Sensing Probe for the Analysis of Zinc Bioavailability Using Caco-2 Cells as an In-Vitro Cellular Model

2020 ◽  
Vol 30 (6) ◽  
pp. 1553-1565
Author(s):  
Sandip Nathani ◽  
Vinod Kumar ◽  
Harcharan S. Dhaliwal ◽  
Debabrata Sircar ◽  
Partha Roy
Author(s):  
Sarah Beschta ◽  
Katja Eubler ◽  
Nancy Bohne ◽  
Ignasi Forne ◽  
Dieter Berg ◽  
...  

AbstractHuman primary granulosa cells (GCs) derived from women undergoing oocyte retrieval can be cultured and used as a cellular model for the study of human ovarian function. In vitro, they change rapidly, initially resembling cells of the preovulatory follicle and then cells of the corpus luteum. They are derived from individual patients, whose different medical history, lifestyle and age lead to heterogeneity. Thus, cells can rarely be ideally matched for cellular experiments or, if available, only in small quantities. We reasoned that cryopreservation of human GCs may be helpful to improve this situation. Previous studies indicated the feasibility of such an approach, but low survival of human GCs was reported, and effects on human GC functionality were only partially evaluated. We tested a slow freezing protocol (employing FCS and DMSO) for human GCs upon isolation from follicular fluid. We compared cryopreserved and subsequently thawed cells with fresh, non-cryopreserved cells from the same patients. About 80% of human GCs survived freezing/thawing. No differences were found in cell morphology, survival rate in culture, or transcript levels of mitochondrial (COX4, OPA1, TOMM20), steroidogenic (CYP11A1, CYP19A1) or cell–cell contact genes (GJA1) between the two groups in cells cultured for 1–5 days. A proteomic analysis revealed no statistically significant change in the abundance of a total of 5962 proteins. The two groups produced comparable basal levels of progesterone and responded similarly to hCG with elevation of progesterone. Taken together, our results show this to be a rapid and readily available method for the cryopreservation of human GCs. We anticipate that it will allow future large-scale experiments and may thereby improve cellular studies with human ovarian cells.


2001 ◽  
Vol 21 (4) ◽  
pp. 277-279
Author(s):  
Hao Youhua ◽  
Wu Xiongwen ◽  
Liang Zhihui ◽  
Xiong Ping ◽  
Jiang Xiaodan ◽  
...  
Keyword(s):  

Reproduction ◽  
2020 ◽  
Vol 160 (2) ◽  
pp. 259-268 ◽  
Author(s):  
Nina Schmid ◽  
Annika Missel ◽  
Stoyan Petkov ◽  
Jan B Stöckl ◽  
Florian Flenkenthaler ◽  
...  

Testicular peritubular cells (TPCs) are smooth muscle-like cells, which form a compartment surrounding the seminiferous tubules. Previous studies employing isolated human testicular peritubular cells (HTPCs) indicated that their roles in the testis go beyond sperm transport and include paracrine and immunological contributions. Peritubular cells from a non-human primate (MKTPCs), the common marmoset monkey, Callithrix jacchus, share a high degree of homology with HTPCs. However, like their human counterparts these cells age in vitro and replicative senescence limits in-depth functional or mechanistic studies. Therefore, a stable cellular model was established. MKTPCs of a young adult animal were immortalized by piggyBac transposition of human telomerase (hTERT), that is, without the expression of viral oncogenes. Immortalized MKTPCs (iMKTPCs) grew without discernable changes for more than 50 passages. An initial characterization revealed typical genes expressed by peritubular cells (androgen receptor (AR), smooth-muscle actin (ACTA2), calponin (CNN1)). A proteome analysis of the primary MKTPCs and the derived immortalized cell line confirmed that the cells almost completely retained their phenotype. To test whether they respond in a similar way as HTPCs, iMKTPCs were challenged with forskolin (FSK) and ATP. As HTPCs, they showed increased expression level of the StAR protein (StAR) after FSK stimulation, indicating steroidogenic capacity. ATP increased the expression of pro-inflammatory factors (e.g. IL1B; CCL7), as it is the case in HTPCs. Finally, we confirmed that iMKTPCs can efficiently be transfected. Therefore, they represent a highly relevant translational model, which allows mechanistic studies for further exploration of the roles of testicular peritubular cells.


2019 ◽  
Vol 25 (5) ◽  
pp. e3162 ◽  
Author(s):  
Federica Tonolo ◽  
Laura Moretto ◽  
Stefania Ferro ◽  
Alessandra Folda ◽  
Valeria Scalcon ◽  
...  

2013 ◽  
Vol 19 (15-16) ◽  
pp. 1665-1674 ◽  
Author(s):  
Mireia Alemany-Ribes ◽  
María García-Díaz ◽  
Marta Busom ◽  
Santi Nonell ◽  
Carlos E. Semino
Keyword(s):  

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Sylvie Rodrigues-Ferreira ◽  
Marina Morel ◽  
Rosana I. Reis ◽  
Françoise Cormier ◽  
Véronique Baud ◽  
...  

Recent studies have highlighted the AT1 receptor as a potential therapeutic target in breast cancer, while the role of the AT2 subtype in this disease has remained largely neglected. The present study describes the generation and characterization of a new cellular model of human invasive breast cancer cells (D3H2LN-AT2) stably expressing high levels of Flag-tagged human AT2 receptor (Flag-hAT2). These cells exhibit high-affinity binding sites for AngII, and total binding can be displaced by the AT2-selective antagonist PD123319 but not by the AT1-selective antagonist losartan. Of interest, high levels of expression of luciferase and green fluorescent protein make these cells suitable for bioluminescence and fluorescence studies in vitro and in vivo. We provide here a novel tool to investigate the AT2 receptor functions in breast cancer cells, independently of AT1 receptor activation.


Gut ◽  
2007 ◽  
Vol 56 (9) ◽  
pp. 1302-1308 ◽  
Author(s):  
C Hourioux ◽  
R Patient ◽  
A Morin ◽  
E Blanchard ◽  
A Moreau ◽  
...  

Materials ◽  
2019 ◽  
Vol 12 (21) ◽  
pp. 3633 ◽  
Author(s):  
Devis Bellucci ◽  
Elena Veronesi ◽  
Valentina Strusi ◽  
Tiziana Petrachi ◽  
Alba Murgia ◽  
...  

A 3D cellular model that mimics the potential clinical application of a biomaterial is here applied for the first time to a bioactive glass, in order to assess its biological potential. A recently developed bioactive glass (BGMS10), whose composition contained strontium and magnesium, was produced in the form of granules and fully investigated in terms of biocompatibility in vitro. Apart from standard biological characterization (Simulated Body Fluid (SBF) testing and biocompatibility as per ISO10993), human bone marrow mesenchymal stromal/stem cells (BM-MSCs) were used to investigate the performance of the bioactive glass granules in an innovative 3D cellular model. The results showed that BGMS10 supported human BM-MSCs adhesion, colonization, and bone differentiation. Thus, bioactive glass granules seem to drive osteogenic differentiation and thus look particularly promising for orthopedic applications, bone tissue engineering and regenerative medicine.


2020 ◽  
Vol 7 ◽  
pp. 1263-1271 ◽  
Author(s):  
Shawn Owiredu ◽  
Abhay Ranganathan ◽  
John C. Greenwood ◽  
Sarah Piel ◽  
Joanna I. Janowska ◽  
...  

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