scholarly journals Controlling fluid flow to improve cell seeding uniformity

2018 ◽  
Author(s):  
Paul M. Reynolds ◽  
Camilla Holzmann Rasmussen ◽  
Mathias Hansson ◽  
Martin Dufva ◽  
Mathis O. Riehle ◽  
...  

AbstractStandard methods for seeding monolayer cell cultures in a multiwell plate or dish do not uniformly distribute cells on the surface. With traditional methods, users find aggregation around the circumference, in the centre, or a combination of the two. This variation is introduced due to the macro scale flow of the cell seeding suspension, and movement of the dish before cells can settle and attach to the surface. Reproducibility between labs, users, and experiments is hampered by this variability in cell seeding. We present a simple method for uniform and user-independent cell seeding using an easily produced uniform cell seeder (UCS) device. This allows precise control of cell density in a reproducible manner. By containing the cell seeding suspension in a defined volume above the culture surface with the UCS, fluctuations in cell density are minimised. Seeding accuracy, as defined by the actual cell density versus the target seeding density is improved dramatically across users with various levels of expertise. We go on to demonstrate the impact of local variation in cell density on the lineage commitment of human embryonic stem cells (hESCs) towards pancreatic endoderm (PE). Variations in the differentiation profile of cells across a culture well closely mirror variations in cell density introduced by seeding method – with the UCS correcting variations in differentiation efficiency. The UCS device provides a simple and reproducible method for uniform seeding across multiple culture systems.

2014 ◽  
Vol 1 (1) ◽  
pp. 31-36 ◽  
Author(s):  
O. Zhukorskiy ◽  
O. Gulay ◽  
V. Gulay ◽  
N. Tkachuk

Aim. To determine the response of the populations of Erysipelothrix rhusiopathiae and Leptospira interrogans pathogenic microorganisms to the impact of broadleaf cattail (Thypha latifolia) root diffusates. Methods. Aqueous solutions of T. latifolia root diffusates were sterilized by vacuum fi ltration through the fi lters with 0.2-micron pore diameter. The experimental samples contained cattail secretions, sterile water, and cultures of E. rhusiopathiae or L. interrogans. The same amount of sterile water, as in the experimental samples, was used for the purpose of control, and the same quantity of microbial cultures was added in it. After exposure, the density of cells in the experimental and control samples was determined. Results. Root diffusates of T. latifolia caused an increase in cell density in the populations of E. rhusiopathiae throughout the whole range of the studied dilutions (1:10–1:10000). In the populations of the 6 studied serological variants of L. interrogans spirochetes (pomona, grippotyphosa, copenhageni, kabura, tarassovi, canicola), the action of broadleaf cattail root diffusates caused the decrease in cell density. A stimulatory effect was marked in the experimental samples of the pollonica serological variant of leptospira. Conclusions. The populations of E. rhusiopathiae and L. interrogans pathogenic microorganisms respond to the allelopathic effect of Thypha latifolia by changing the cell density. The obtained results provide the background to assume that broadleaf cattail thickets create favorable conditions for the existence of E. rhusiopathiae pathogen bacteria. The reduced cell density of L. interrogans in the experimental samples compared to the control samples observed under the infl uence of T. latifolia root diffusates suggests that reservoirs with broadleaf cattail thickets are marked by the unfavorable conditions for the existence of pathogenic leptospira (except L. pollonica).


2021 ◽  
pp. 0021955X2110137
Author(s):  
Yao Dou ◽  
Denis Rodrigue

In this study, foamed recycled high density polyethylene (rHDPE) parts were produced by rotational molding using different concentration (0 to 1% wt.) of a chemical blowing agent (CBA) based on azodicarbonamide. From the samples produced, a complete morphological, thermal and mechanical characterization was performed. The morphological analysis showed a gradual increase in the average cell size, while the cell density firstly increased and then decreased with increasing CBA content. As expected, increasing the CBA content decreased the foam density as well as the thermal conductivity. Although increasing the CBA content decreased both tensile and flexural properties, the impact strength showed a similar trend as the cell density with an optimum CBA content around 0.1% wt. Finally, neat rHDPE samples were also produced by compression molding. The results showed negligible differences between the rotomolded and compression molded properties indicating that optimal rotomolding conditions were selected. These results confirm the possibility of using 100% recycled polymers to produce rotomolded foam parts.


2021 ◽  
Vol 22 (2) ◽  
pp. 501
Author(s):  
Kateřina Skopalová ◽  
Katarzyna Anna Radaszkiewicz ◽  
Věra Kašpárková ◽  
Jaroslav Stejskal ◽  
Patrycja Bober ◽  
...  

The active role of biomaterials in the regeneration of tissues and their ability to modulate the behavior of stem cells in terms of their differentiation is highly advantageous. Here, polypyrrole, as a representantive of electro-conducting materials, is found to modulate the behavior of embryonic stem cells. Concretely, the aqueous extracts of polypyrrole induce neurogenesis within embryonic bodies formed from embryonic stem cells. This finding ledto an effort to determine the physiological cascade which is responsible for this effect. The polypyrrole modulates signaling pathways of Akt and ERK kinase through their phosphorylation. These effects are related to the presence of low-molecular-weight compounds present in aqueous polypyrrole extracts, determined by mass spectroscopy. The results show that consequences related to the modulation of stem cell differentiation must also be taken into account when polypyrrole is considered as a biomaterial.


2021 ◽  
Vol 13 (10) ◽  
pp. 1927
Author(s):  
Fuqin Li ◽  
David Jupp ◽  
Thomas Schroeder ◽  
Stephen Sagar ◽  
Joshua Sixsmith ◽  
...  

An atmospheric correction algorithm for medium-resolution satellite data over general water surfaces (open/coastal, estuarine and inland waters) has been assessed in Australian coastal waters. In situ measurements at four match-up sites were used with 21 Landsat 8 images acquired between 2014 and 2017. Three aerosol sources (AERONET, MODIS ocean aerosol and climatology) were used to test the impact of the selection of aerosol optical depth (AOD) and Ångström coefficient on the retrieved accuracy. The initial results showed that the satellite-derived water-leaving reflectance can have good agreement with the in situ measurements, provided that the sun glint is handled effectively. Although the AERONET aerosol data performed best, the contemporary satellite-derived aerosol information from MODIS or an aerosol climatology could also be as effective, and should be assessed with further in situ measurements. Two sun glint correction strategies were assessed for their ability to remove the glint bias. The most successful one used the average of two shortwave infrared (SWIR) bands to represent sun glint and subtracted it from each band. Using this sun glint correction method, the mean all-band error of the retrieved water-leaving reflectance at the Lucinda Jetty Coastal Observatory (LJCO) in north east Australia was close to 4% and unbiased over 14 acquisitions. A persistent bias in the other strategy was likely due to the sky radiance being non-uniform for the selected images. In regard to future options for an operational sun glint correction, the simple method may be sufficient for clear skies until a physically based method has been established.


2003 ◽  
Vol 47 (11) ◽  
pp. 85-92 ◽  
Author(s):  
E. Cotteux ◽  
P. Duchene

The bulking that occurs in biological wastewater treatment plants using activated sludge is very often controlled by the injection of sodium hypochlorite into the return activated sludge (RAS) stream. In the present study undertaken at two pilot plants fed with synthetic wastewater, the impact of the pass frequency of the sludge at the chlorine dosing point on the nitrifying flora is analysed. The pass frequency is one for the pilot plant 1 and two for the pilot plant 2. A dose of chlorine of 4.85 ± 0.05 g/kg/MLVSS per day was applied at both pilots. The preservative effect on nitrifying activity of the lowest concentration of chlorine at the dosing point and therefore of the highest pass frequency was evidenced. Among other tools, a simple method of measurement of the oxygen uptake rate enabled us to monitor the effect of chlorination on nitrification before recording an increase in the ammonia concentration in the bulking.


2018 ◽  
Author(s):  
Robert Reinecke ◽  
Laura Foglia ◽  
Steffen Mehl ◽  
Tim Trautmann ◽  
Denise Cáceres ◽  
...  

Abstract. To quantify water flows between groundwater (GW) and surface water (SW) as well as the impact of capillary rise on evapotranspiration by global hydrological models (GHMs), it is necessary to replace the bucket-like linear GW reservoir model typical for hydrological models with a fully integrated gradient-based GW flow model. Linear reservoir models can only simulate GW discharge to SW bodies, provide no information on the location of the GW table and assume that there is no GW flow among grid cells. A gradient-based GW model simulates not only GW storage but also hydraulic head, which together with information on SW table elevation enables the quantification of water flows from GW to SW and vice versa. In addition, hydraulic heads are the basis for calculating lateral GW flow among grid cells and capillary rise. G3M is a new global gradient-based GW model with a spatial resolution of 5' that will replace the current linear GW reservoir in the 0.5° WaterGAP Global Hydrology Model (WGHM). The newly developed model framework enables in-memory coupling to WGHM while keeping overall runtime relatively low, allowing sensitivity analyses and data assimilation. This paper presents the G3M concept and specific model design decisions together with results under steady-state naturalized conditions, i.e. neglecting GW abstractions. Cell-specific conductances of river beds, which govern GW-SW interaction, were determined based on the 30'' steady-state water table computed by Fan et al. (2013). Together with an appropriate choice for the effective elevation of the SW table within each grid cell, this enables a reasonable simulation of drainage from GW to SW such that, in contrast to the GW model of de Graaf et al. (2015, 2017), no additional drainage based on externally provided values for GW storage above the floodplain is required in G3M. Comparison of simulated hydraulic heads to observations around the world shows better agreement than de Graaf et al. (2015). In addition, G3M output is compared to the output of two established macro-scale models for the Central Valley, California, and the continental United States, respectively. As expected, depth to GW table is highest in mountainous and lowest in flat regions. A first analysis of losing and gaining rivers and lakes/wetlands indicates that GW discharge to rivers is by far the dominant flow, draining diffuse GW recharge, such that lateral flows only become a large fraction of total diffuse and focused recharge in case of losing rivers and some areas with very low GW recharge. G3M does not represent losing rivers in some dry regions. This study presents the first steps towards replacing the linear GW reservoir model in a GHM while improving on recent efforts, demonstrating the feasibility of the approach and the robustness of the newly developed framework.


2018 ◽  
Vol 33 (4) ◽  
pp. 576-589 ◽  
Author(s):  
Devika M Varma ◽  
Michelle S DiNicolas ◽  
Steven B Nicoll

Low back pain is a major cause for disability and is closely linked to intervertebral disc degeneration. Mechanical and biological dysfunction of the nucleus pulposus in the disc has been found to initiate intradiscal degenerative processes. Replacing or enriching the diseased nucleus pulposus with an injectable, stem cell-laden biomaterial that mimics its material properties can provide a minimally invasive strategy for biological and structural repair of the tissue. In this study, injectable, in situ-gelling carboxymethylcellulose hydrogels were developed for nucleus pulposus tissue engineering using encapsulated human marrow-derived mesenchymal stromal cells (hMSCs). With the goal of obtaining robust extracellular matrix deposition and faster construct maturation, two cell-seeding densities, 20 × 106 cells/ml and 40 × 106 cells/ml, were examined. The constructs were fabricated using a redox initiation system to yield covalently crosslinked, cell-seeded hydrogels via radical polymerization. Chondrogenic culture of the hydrogels over 35 days exhibited high cell viability along with deposition of proteoglycan and collagen-rich extracellular matrix, and mechanical and swelling properties similar to native human nucleus pulposus. Further, the matrix production and distribution in the carboxymethylcellulose hydrogels was found to be strongly influenced by hMSC-seeding density, with the lower cell-seeding density yielding a more favorable nucleus pulposus-specific matrix phenotype, while the rate of construct maturation was less dependent on the cell-seeding density. These findings are the first to demonstrate the utility of redox-polymerized carboxymethylcellulose hydrogels as hMSC carriers for potential minimally invasive treatment strategies for nucleus pulposus replacement.


2018 ◽  
Vol 2018 ◽  
pp. 1-14
Author(s):  
José Carlos Ortiz-Bayliss ◽  
Ivan Amaya ◽  
Santiago Enrique Conant-Pablos ◽  
Hugo Terashima-Marín

When solving constraint satisfaction problems (CSPs), it is a common practice to rely on heuristics to decide which variable should be instantiated at each stage of the search. But, this ordering influences the search cost. Even so, and to the best of our knowledge, no earlier work has dealt with how first variable orderings affect the overall cost. In this paper, we explore the cost of finding high-quality orderings of variables within constraint satisfaction problems. We also study differences among the orderings produced by some commonly used heuristics and the way bad first decisions affect the search cost. One of the most important findings of this work confirms the paramount importance of first decisions. Another one is the evidence that many of the existing variable ordering heuristics fail to appropriately select the first variable to instantiate. Another one is the evidence that many of the existing variable ordering heuristics fail to appropriately select the first variable to instantiate. We propose a simple method to improve early decisions of heuristics. By using it, performance of heuristics increases.


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