scholarly journals NanoPASS: an easy-to-use user interface for nanoparticle dosimetry with the 3DSDD model

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Falko Frenzel ◽  
Laura König-Mattern ◽  
Valerie Stock ◽  
Linn Voss ◽  
Maxi B. Paul ◽  
...  
Keyword(s):  
Cell Culture ◽  
In Silico ◽  
Physicochemical Characteristics ◽  
Time Frame ◽  
Culture Conditions ◽  
Sedimentation Behavior ◽  
A Cell ◽  
Correct Application ◽  
Cell Culture Conditions

Abstract Nanoparticles exhibit a specific diffusion and sedimentation behavior under cell culture conditions as used in nantoxicological in vitro testing. How a particular particle suspension behaves depends on the particular physicochemical characteristics of the particles and the cell culture system. Only a fraction of the nanoparticles applied to a cell culture will thus reach the cells within a given time frame. Therefore, dosimetric calculations are essential not only to determine the exact fraction of nanoparticles that has come into contact with the cells, but also to ensure experimental comparability and correct interpretation of results, respectively. Yet, the use of published dosimetry models is limited. Not the least because the correct application of these in silico tools usually requires bioinformatics knowledge, which often is perceived a hurdle. Moreover, not all models are freely available and accessible. In order to overcome this obstacle, we have now developed an easy-to-use interface for our recently published 3DSDD dosimetry model, called NanoPASS (NanoParticle Administration Sedimentation Simulator). The interface is freely available to all researchers. It will facilitate the use of in silico dosimetry in nanotoxicology and thus improve interpretation and comparability of in vitro results in the field.

Laser microstructured scaffolds for tissue engineering under dynamic cell culture conditions

2020 ◽  
Author(s):  
Ελευθερία Μπαμπαλιάρη
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
Culture Conditions ◽  
Dynamic Cell ◽  
Cell Culture Conditions

Παρόλο που το περιφερικό νευρικό σύστημα εμφανίζει υψηλότερο ρυθμό αναγέννησης από εκείνο του κεντρικού νευρικού συστήματος μέσω αυθόρμητης αναγέννησης μετά από έναν τραυματισμό, η καθοδηγούμενη αξονική νευρική αναγέννηση και η λειτουργική αποκατάσταση είναι αρκετά σπάνια. Συνεπώς, η ανάπτυξη επιτυχημένων μεθόδων για την καθοδήγηση της νευρικής ανάπτυξης, «in vitro», είναι υψίστης σημασίας. Έχει αναφερθεί λεπτομερώς ότι η τοπογραφία του υποστρώματος επηρεάζει την ανάπτυξη, τον προσανατολισμό και τη διαφοροποίηση των νευρικών κυττάρων. Ωστόσο, η συνδυασμένη δράση της διατμητικής τάσης και της τοπογραφίας του υποστρώματος στην νευρική ανάπτυξη έχει ελάχιστα μελετηθεί, παρόλο που οι διατμητικές τάσεις είναι ευρέως γνωστό ότι διαδραματίζουν καθοριστικό ρόλο στην οργάνωση, ανάπτυξη και λειτουργία των ιστών. Σε αυτή τη μελέτη, ένα σύστημα μικροροών ακριβούς ελεγχόμενης ροής με συγκεκριμένους ειδικά σχεδιασμένους θαλάμους, που ενσωματώνουν μικροδομημένα υποστρώματα λέιζερ, αναπτύχθηκε για να μελετηθεί η συνδυασμένη δράση της διατμητικής τάσης και της τοπογραφίας υποστρώματος στην ανάπτυξη, στον προσανατολισμό, στην επιμήκυνση και στη διαφοροποίηση νευρικών κυττάρων. Πολυμερικά μικροδομημένα υποστρώματα, με ελεγχόμενη γεωμετρία και κανονικότητα μοτίβου, κατασκευάστηκαν με χρήση υπερβραχέων παλμών λέιζερ. Πραγματοποιήθηκε συγκριτική μελέτη μεταξύ στατικών και δυναμικών κυτταρικών καλλιεργειών για να αξιολογηθεί η συνεργατική ή ανταγωνιστική επίδραση της διατμητικής τάσης και της τοπογραφίας στη συμπεριφορά των νευρικών κυττάρων. Τα αποτελέσματα της κυτταρικής καλλιέργειας συμπληρώθηκαν με υπολογιστικές προσομοιώσεις ροής με σκοπό τον ακριβή υπολογισμό των αντίστοιχων τιμών διατμητικής τάσης.

scholarly journals In Vitro Matrix Assembly Induced by Critical Assembly Concentration (CAC)

2002 ◽  
Vol 50 (11) ◽  
pp. 1537-1541 ◽  
Author(s):  
Mark E. Lauer ◽  
Kevin J. McCarthy
Keyword(s):  
Cell Culture ◽  
Cell Types ◽  
Culture Conditions ◽  
Protein Diffusion ◽  
Matrix Assembly ◽  
Physical Limitations ◽  
Reichert's Membrane ◽  
Immortalized Cell ◽  
Cell Culture Conditions

L-2 cells are an immortalized cell line derived from yolk sac parietal endoderm cells, which are responsible for the production of Reichert's membrane, a thick basement membrane produced during rat gestation. Although the L-2 cells secrete all the major components of the basal lamina, they do not assemble a robust matrix in cell culture. We hypothesized that the reason L-2 cells fail to assemble a matrix in cell culture is because the concentrations of matrix components necessary for this matrix assembly do not reach a critical association concentration (CAC) under standard cell culture conditions. To limit the diffusion of secreted molecules while maintaining a nutrient-rich environment for the cells to thrive, we developed a technique that uses a dialysis membrane to limit protein diffusion in a 2-well plate format. This technique permits L-2 cells to assemble a robust matrix in as little as 24 hr that continues to be formed for at least 72 hr. This technique may address some of the physical limitations imposed by cell culture and could be readily applied to other cell types and medium conditions.

scholarly journals Mitochondrial and metabolic remodeling in human skin fibroblasts in response to glucose availability

2021 ◽  
Author(s):  
Cláudio F. Costa ◽  
Sónia A. Pinho ◽  
Sonia L.C. Pinho ◽  
Inês Miranda-Santos ◽  
Olivia Bagshaw ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Model ◽  
Complete Removal ◽  
Culture Conditions ◽  
Dermal Fibroblasts ◽  
Physiological Level ◽  
Atp Production ◽  
Metabolic Remodeling ◽  
Cell Culture Conditions

AbstractCell culture conditions highly influence cell metabolism in vitro. This is relevant for preclinical assays, for which fibroblasts are an interesting cell model, with applications in regenerative medicine, diagnostics and therapeutic development for personalized medicine as well as in the validation of ingredients for cosmetics. Given these cells’ short lifespan in culture, we aimed to identify the best cell culture conditions and promising markers to study mitochondrial health and stress in Normal Human Dermal Fibroblasts (NHDF). We tested the effect of reducing glucose concentration in the cell medium from high glucose (HGm) to a more physiological level (LGm), or its complete removal and replacement by galactose (OXPHOSm), always in the presence of glutamine and pyruvate. We have demonstrated that only with OXPHOSm it was possible to observe the selective inhibition of mitochondrial ATP production. This reliance on mitochondrial ATP was accompanied by changes in oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), oxidation of citric acid cycle substrates, fatty acids, lactate and other substrates, mitochondrial network extension and polarization and changes in several key transcripts related to energy metabolism. We also evaluated the relevance of galactose, glutamine and pyruvate for OXPHOS stimulation, by comparing OCR and ECAR in the presence or absence of these substrates. Galactose and pyruvate seem to be important, but redundant, to promote OXPHOS, whereas glutamine was essential. We concluded that LGm does not promote significant metabolic changes but the short-term adaptation to OXPHOSm is ideal for studying mitochondrial health and stress in NHDF.Author ContributionsCC, SAP, SLCP and IMS performed experiments. TCO and PJO designed research and acquired funding. JS, and OB analyzed data. CC and TCO analyzed data and wrote the paper. All authors contributed to the final version of the manuscript.

scholarly journals An Improved Method of Maintaining Primary Murine Cardiac Fibroblasts in Two-Dimensional Cell Culture

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Natalie M. Landry ◽  
Sunil G. Rattan ◽  
Ian M. C. Dixon
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Cardiac Fibroblasts ◽  
Smooth Muscle Actin ◽  
Growth Factor Receptor ◽  
Culture Conditions ◽  
Quiescent State ◽  
Alpha Smooth Muscle Actin ◽  
Cell Culture Conditions

Abstract Primary cardiac fibroblasts are notoriously difficult to maintain for extended periods of time in cell culture, due to the plasticity of their phenotype and sensitivity to mechanical input. In order to study cardiac fibroblast activation in vitro, we have developed cell culture conditions which promote the quiescent fibroblast phenotype in primary cells. Using elastic silicone substrata, both rat and mouse primary cardiac fibroblasts could be maintained in a quiescent state for more than 3 days after isolation and these cells showed low expression of myofibroblast markers, including fibronectin extracellular domain A, non-muscle myosin IIB, platelet-derived growth factor receptor-alpha and alpha-smooth muscle actin. Gene expression was also more fibroblast-like vs. that of myofibroblasts, as Tcf21 was significantly upregulated, while Fn1-EDA, Col1A1 and Col1A2 were markedly downregulated. Cell culture conditions (eg. serum, nutrient concentration) are critical for the control of temporal fibroblast proliferation. We propose that eliminating mechanical stimulus and limiting the nutrient content of cell culture media can extend the quiescent nature of primary cardiac fibroblasts for physiological analyses in vitro.

scholarly journals Recent advances in microfluidic technologies for cell-to-cell interaction studies

Lab on a Chip ◽  
2018 ◽  
Vol 18 (2) ◽  
pp. 249-270 ◽  
Author(s):  
Mario Rothbauer ◽  
Helene Zirath ◽  
Peter Ertl
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Cell Signalling ◽  
Cell Interaction ◽  
Culture Conditions ◽  
Diagnostic Tools ◽  
Biological Processes ◽  
Dynamic Cell ◽  
Cell Culture Conditions

Microfluidic cell cultures are ideally positioned to become the next generation ofin vitrodiagnostic tools for biomedical research, where key biological processes such as cell signalling and dynamic cell-to-cell interactions can be reliably analysed under reproducible physiological cell culture conditions.

scholarly journals The Roles of Insulin-Like Growth Factors in Mesenchymal Stem Cell Niche

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Amer Youssef ◽  
Doaa Aboalola ◽  
Victor K. M. Han
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Stem Cell ◽  
Growth Factors ◽  
Culture Conditions ◽  
Self Renewal ◽  
Adult Mesenchymal Stem Cells ◽  
Cell Culture Conditions ◽  
Insulin Like Growth Factors

Many tissues contain adult mesenchymal stem cells (MSCs), which may be used in tissue regeneration therapies. However, the MSC availability in most tissues is limited which demands expansion in vitro following isolation. Like many developing cells, the state of MSCs is affected by the surrounding microenvironment, and mimicking this natural microenvironment that supports multipotent or differentiated state in vivo is essential to understand for the successful use of MSC in regenerative therapies. Many researchers are, therefore, optimizing cell culture conditions in vitro by altering growth factors, extracellular matrices, chemicals, oxygen tension, and surrounding pH to enhance stem cells self-renewal or differentiation. Insulin-like growth factors (IGFs) system has been demonstrated to play an important role in stem cell biology to either promote proliferation and self-renewal or enhance differentiation onset and outcome, depending on the cell culture conditions. In this review, we will describe the importance of IGFs, IGF-1 and IGF-2, in development and in the MSC niche and how they affect the pluripotency or differentiation towards multiple lineages of the three germ layers.

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