Media Recipes Research Articles

Highly effective osteogenic conversion of mesenchymal Stem Cells by Cell Rev™ Supreme Osteo

Background & Aim Differentiation potentials of multipotent mesenchymal stem cells (MSCs) are one of the two primary hallmarks of MSCs. Media recipes have been formulated to enhance growth properties but often at the expense of differentiation potentials. Two major factors that are responsible for blunting MSC differentiation potential include the expansion as well as the differentiation media. The former can significantly hurdle MSCs properties especially in long term expanded MSCs. We aim to solve this major setback of MSCs quality; here we focus on osteogenic differentiation as bone repair by MSCs cell therapy is at the forefront of clinic use. Methods, Results & Conclusion Osteogenic differentiation is accounted by two cellular milieu: 1) the media that MSCs have been cultivated with and 2) the differentiation media to convert the MSCs to osteoblasts. We examine these issues experimentally by: comparing conversion rate and quality of osteogenic differentiation of A) MSCs grown on FBS derived standard media (STD) and MSCs has grown on Cell Rev™ MSC ExHQ 2.0 media, and B) comparing effectiveness of osteogenic differentiation of the MSCs that were cultivated on either industry-standard osteogenic differentiation kit (STD) or Cell Rev™ Supreme Osteo. First, Cell Rev™ Supreme Osteo increased MSCs conversion rates from 10% to 70% even if the MSCs were cultured in STD media. More interestingly, Cell Rev™ Supreme Osteo was able to achieve a 100% differentiation when the MSCs were expanded from Cell Rev™ MSC ExHQ 2.0. The results show synergistic effects of retaining high-level differentiation potential from maintaining MSCs in Cell Rev™ MSC ExHQ 2.0, and that contribute to the most effective osteoblast conversion when cultivated with Cell Rev™ Supreme Osteo. These results illustrate that Cell Rev™ Supreme Osteo is highly effective in converting MSCs to osteoblasts regardless of poor expansion media. This solution of complete osteoblast conversion is important for the safety issue of bone repair in clinical practices because it eliminates the risks of MSCs turning into unwanted types of cells.

H-Aquil: a chemically defined cell culture medium for trace metal studies in Vibrios and other marine heterotrophic bacteria.

A variety of trace metals, including prominently iron (Fe) are necessary for marine microorganisms. Chemically defined medium recipes have been used for several decades to study phytoplankton, but similar methods have not been adopted as widely in studies of marine heterotrophic bacteria. Medium recipes for these organisms frequently include tryptone, casamino acids, as well as yeast and animal extracts. These components introduce unknown concentrations of trace elements and organic compounds, complicating metal speciation. Minimal medium recipes utilizing known carbon and nitrogen sources do exist but often have high background trace metal concentrations. Here we present H-Aquil, a version of the phytoplankton medium Aquil adapted for marine heterotrophic bacteria. This medium consists of artificial seawater supplemented with a carbon source, phosphate, amino acids, and vitamins. As in Aquil, trace metals are controlled using the synthetic chelator EDTA. We also address concerns of EDTA toxicity, showing that concentrations up to 100µM EDTA do not lead to growth defects in the copiotrophic bacterium Vibrio harveyi or the oligotrophic bacterium Candidatus Pelagibacter ubique HTCC1062, a member of the SAR11 clade. H-Aquil is used successfully to culture species of Vibrio, Phaeobacter, and Silicibacter, as well as several environmental isolates. We report a substantial decrease in growth rate between cultures grown with or without added Fe, making the medium suitable for conducting Fe-limitation studies in a variety of marine heterotrophic bacteria.

Exploring factors affecting anther culture in rice (Oryza sativa L.)

Anther culture is an important in vitro culture technique for production of double haploids. It is largely species and genotype-specific. Asian cultivated rice (O. sativa, ssp. indica) is recalcitrant to anther culture which limits its practical application in rice breeding. Several researchers tried to optimize the media recipes and culture techniques for callus induction and plantlet regeneration. Negligible response to callus induction and recovery of higher frequency of albino plants are the major hindrance for use of the technique in crop improvement. Shortening the culture period and sexual hybridization of rice sub-groups (japonica/indica) may be adopted to improve green plant regeneration from anther-derived callus in indica rice. However, genetic transformation technology using individual genes related to different aspects of anther culture could be a more direct approach. Besides, the role of genotypes, physiological status of donor plant, developmental stage of pollen, pre-treatment, culture media, phytohormones and culture conditions for successful anther culture have been discussed. Key words: Anther culture, factors for anther culture, double haploids, genetic improvement, rice.

Increasing the buffering capacity of minimal media leads to higher protein yield.

We describe a general and simple modification to the standard M9 minimal medium recipe that leads to an approximate twofold increase in the yield of heterologously expressed proteins in Escherichia coli BL21(DE3) bacteria. We monitored the growth of bacteria transformed with plasmids for three different test proteins in five minimal media with different concentrations of buffering salts and/or initial media pH. After purification of the over-expressed proteins, we found a clear correlation between the protein yield and change in media pH over time, where the minimal media that were the most buffered and therefore most resistant to change in pH produced the most protein. And in all three test protein cases, the difference in yield was nearly twofold between the best and worst buffering media. Thus, we propose that increasing the buffering capacity of M9 minimal media will generally lead to a similar increase for most of the proteins currently produced by this standard protein expression protocol. Moreover, we have qualitatively found that this effect also extends to deuterated M9 minimal media growths, which could lead to significant cost savings in these preparations.

Anther culture in rice: Progress and breeding perspective

Anther culture is an important biotechnological tool for the rapid recovery of fixed breeding lines in rice. The method is largely species- and genotype-specific. Indica rice is known to have a recalcitrant genetic background. Oryza glaberrima responds better to the anther culture than O. sativa while japonica sub-group is more responsive to microspore embryogenesis than Indica types. Media recipes and culture techniques have been standardized for callus induction and green plantlet regeneration using Indica and Japonica varieties as well as their inter se hybrids. Additive effects seem more important than dominant effects of the genes concerned for callus induction while preponderance of dominant effects is reported for regeneration response. Sexual hybridization of rice sub-groups (Japonica x Indica) followed by the selection of favourable QTL for green plant regeneration may be adopted to improve plant regeneration from anther-derived callus in Indica rice. However, genetic transformation technology using individual genes related to different aspects of anther culture could be a more direct approach. This paper reviews the progress and breeding perspective of anther culture in rice for genetic improvement of agronomic- and stress-related traits using double haploid breeding.

Changes in phenotype and differentiation potential of human mesenchymal stem cells aging in vitro

BackgroundAdult mesenchymal stem cells (MSCs) hold great promise for regenerative medicine because of their self-renewal, multipotency, and trophic and immunosuppressive effects. Due to the rareness and high heterogeneity of freshly isolated MSCs, extensive in-vitro passage is required to expand their populations prior to clinical use; however, senescence usually accompanies and can potentially affect MSC characteristics and functionality. Therefore, a thorough characterization of the variations in phenotype and differentiation potential of in-vitro aging MSCs must be sought.MethodsHuman bone marrow-derived MSCs were passaged in vitro and cultivated with either DMEM-based or αMEM-based expansion media. Cells were prepared for subculture every 10 days up to passage 8 and were analyzed for cell morphology, proliferative capacity, and surface marker expression at the end of each passage. The gene expression profile and adipogenic and osteogenic differentiation capability of MSCs at early (passage 4) and late (passage 8) passages were also evaluated.ResultsIn-vitro aging MSCs gradually lost the typical fibroblast-like spindle shape, leading to elevated morphological abnormality and inhomogeneity. While the DMEM-based expansion medium better facilitated MSC proliferation in the early passages, the cell population doubling rate reduced over time in both DMEM and αMEM groups. CD146 expression decreased with increasing passage number only when MSCs were cultured under the DMEM-based condition. Senescence also resulted in MSCs with genetic instability, which was further regulated by the medium recipe. Regardless of the expansion condition, MSCs at both passages 4 and 8 could differentiate into adipocyte-like cells whereas osteogenesis of aged MSCs was significantly compromised. For osteogenic induction, use of the αMEM-based expansion medium yielded longer osteogenesis and better quality.ConclusionsHuman MSCs subjected to extensive in-vitro passage can undergo morphological, phenotypic, and genetic changes. These properties are also modulated by the medium composition employed to expand the cell populations. In addition, adipogenic potential may be better preserved over osteogenesis in aged MSCs, suggesting that MSCs at early passages must be used for osteogenic differentiation. The current study presents valuable information for future basic science research and clinical applications leading to the development of novel MSC-based therapeutic strategies for different diseases.

Liquid fermentation of Colletotrichum truncatum UFU 280, a potential mycoherbicide for beggartick

One isolate of Colletotrichum truncatum was found recently causing severe anthracnose symptoms and leading to the death of beggartick (Bidens pilosa L. and Bidens subalternans DC.), one of the major weeds of Brazilian agriculture. This isolate, namely UFU 280, was selected for development of a mycoherbicide against this weed. Associated of beggartick, one of the, Here, results of a preliminary attempt to develop a protocol for mass production of inoculum (conidia) and fungal biomass of C. truncatum was performed aimed at paving the way for greenhouse and field evaluations of this biocontrol candidate. Isolates of Colletotrichum spp. have been successfully produced in the past to serve as the active ingredients of mycoherbicides. The method of choice has been the production of propagules through liquid fermentation. We assessed the effect of several options of liquid media recipes, type of seeding of medium, pH levels, incubation lengths, incubation temperatures and agitation speeds on the shaking speed on the concentration of conidia obtained per volume of medium. Additionally, a possible effect of the kind of medium utilized over the virulence of the inoculum was also evaluated through an inoculation study. We found that an adequate amount of conidia of C. truncatum (isolate - UFU 280) can be obtained in ME liquid culture medium, adjusted to a pH of 9.0, seeded with a conidial suspension and incubated for 6 days, under a regime of orbital shaking of 150 rpm, at temperatures ranging from 20 to 25 °C. Mortality of beggartick plants using conidia produced in different liquid culture media was of 100%.

Anther Culture for Double Haploid Breeding in Rice-a Way Forward

Anther culture is an important biotechnological tool for quick recovery of fixed breeding lines in rice. It is largely species and genotype-specific. O. glaberrima responds better to anther culture than O. sativa; and japonica sub-group is more responsive to microspore embryogenesis than indica types. Media recipes and culture techniques have been standardized for callus induction and green plantlet regeneration using indica and japonica varieties as well as their inter se hybrids. Besides, the genetic basis and scope of anther culture for double haploid breeding in rice have been discussed.

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology

A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of bioprocess conditions. One important aspect is the improvement of cultivation medium to provide an optimal environment for microbial formation of the product of interest. It is well accepted that the media composition can dramatically influence overall bioprocess performance. Nutrition medium optimization is known to improve recombinant protein production with microbial systems and thus, this is a rewarding step in bioprocess development. However, very often standard media recipes are taken from literature, since tailor-made design of the cultivation medium is a tedious task that demands microbioreactor technology for sufficient cultivation throughput, fast product analytics, as well as support by lab robotics to enable reliability in liquid handling steps. Furthermore, advanced mathematical methods are required for rationally analyzing measurement data and efficiently designing parallel experiments such as to achieve optimal information content. The generic nature of the presented protocol allows for easy adaption to different lab equipment, other expression hosts, and target proteins of interest, as well as further bioprocess parameters. Moreover, other optimization objectives like protein production rate, specific yield, or product quality can be chosen to fit the scope of other optimization studies. The applied Kriging Toolbox (KriKit) is a general tool for Design of Experiments (DOE) that contributes to improved holistic bioprocess optimization. It also supports multi-objective optimization which can be important in optimizing both upstream and downstream processes.

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology.

A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of bioprocess conditions. One important aspect is the improvement of cultivation medium to provide an optimal environment for microbial formation of the product of interest. It is well accepted that the media composition can dramatically influence overall bioprocess performance. Nutrition medium optimization is known to improve recombinant protein production with microbial systems and thus, this is a rewarding step in bioprocess development. However, very often standard media recipes are taken from literature, since tailor-made design of the cultivation medium is a tedious task that demands microbioreactor technology for sufficient cultivation throughput, fast product analytics, as well as support by lab robotics to enable reliability in liquid handling steps. Furthermore, advanced mathematical methods are required for rationally analyzing measurement data and efficiently designing parallel experiments such as to achieve optimal information content.The generic nature of the presented protocol allows for easy adaption to different lab equipment, other expression hosts, and target proteins of interest, as well as further bioprocess parameters. Moreover, other optimization objectives like protein production rate, specific yield, or product quality can be chosen to fit the scope of other optimization studies. The applied Kriging Toolbox (KriKit) is a general tool for Design of Experiments (DOE) that contributes to improved holistic bioprocess optimization. It also supports multi-objective optimization which can be important in optimizing both upstream and downstream processes.

Reuse of rejuvenated media during laboratory and pilot scale cultivation of Nannochloropsis sp.

Bulk cultivation of microalgae in large scale production systems is subject to high production costs. Recycling of growth medium contributes to reducing water, nutrient and salt expenditure, and hence to increase both the economic viability and sustainability of microalgae production systems. In this study it was shown that, upon algae harvesting by means of centrifugation, spent broth from a Nannochloropsis sp. production system can be treated by means of microfiltration and reapplied as rejuvenated growth medium without negatively influencing subsequent growth. During recirculation of growth medium, nutrients consumption rates were documented and used to adjust the medium recipe to enable long term medium recirculation. An 80% recycle rate could be maintained for 167days during a course of ten production runs, translating into a decrease in nutrient, salt and water expenditure of 44%, 74% and 80%, respectively. The potential for long term reuse of medium for monocultures of multiple economically interesting species should be assessed in future research to evaluate the broad applicability of medium re-use in microalgae cultivation.

Prevention of melanin formation during aryl alcohol oxidase production under growth-limited conditions using an Aspergillus nidulans cell factory

An Aspergillus nidulans cell factory was genetically engineered to produce an aryl alcohol oxidase (AAO). The cell factory initiated production of melanin when growth-limited conditions were established using stationary plates and shaken flasks. This phenomenon was more pronounced when the strain was cultured in a trickle bed reactor (TBR). This study investigated different approaches to reduce melanin formation in fungal mycelia and liquid medium in order to increase the enzyme production yield. Removal of copper from the medium recipe reduced melanin formation in agar cultures and increased enzyme activities by 48% in agitated liquid cultures. Copper has been reported as a key element for tyrosinase, an enzyme responsible for melanin production. Ascorbic acid (0.44g/L) stopped melanin accumulation, did not affect growth parameters and resulted in AAO activity that was more than two-fold greater than a control treatment with no ascorbic acid.

Determining Optimum Calcium and Magnesium Sources and Rates for Home Gardeners Growing Vegetables in Potting Media Using Alkaline Irrigation Water

Home gardeners living in areas with alkaline water sources do not have easy or economically affordable means of acidifying irrigation water for vegetable production. One solution for achieving optimal vegetable yields using alkaline irrigation water is to grow the vegetables in a modified medium. To date, no medium on the retail market suits such growing needs. Therefore, medium recipes with varied levels (0, 4, or 8 lb/yard3) and sources of calcium [dolomitic lime, calcium sulfate (CaSO4)] and magnesium [dolomitic lime, magnesium sulfate (MgSO4)] were tested using an alkaline irrigation on ‘Oakleaf’ lettuce (Lactuca sativa), ‘Earliana’ and ‘Salad Delight’ cabbage (Brassica oleracea var. capitata), and ‘Snow Crown’ cauliflower (Brassica oleracea var. botrytis) crops. Additionally, crops were grown in two environments, under a high tunnel and on a nursery yard. High tunnel and nursery yard sites were used to test media performances in the presence of, and eliminating, rainwater to simulate container-grown vegetables growing in both a home garden situation and a commercial greenhouse production situation. The base mix of all media treatments in the study was 80 bark : 20 peat and fertilized with 12 lb/yard3 slow-release fertilizer at a rate of 1.8 lb/yard3 nitrogen (N), 0.5 lb/yard3 phosphorus (P), and 1 lb/yard3 potassium (K). This initial fertilizer application was incorporated to each medium before filling containers. Four treatments were tested against a commercially available medium, industry standard (IS) treatment (a commercially available bagged medium), and a control medium [treatment C (no supplemental calcium or magnesium fertilizer)] by supplementing the base mix with the following fertilizer levels: 4 lb/yard3 each of CaSO4 and MgSO4 (treatment 1); 4 lb/yard3 dolomitic lime (treatment 2); 4 lb/yard3 each of dolomitic lime, CaSO4, and MgSO4 (treatment 3); 8 lb/yard3 dolomitic lime (treatment 4). Media treatments 1 through 4 outperformed the IS and C media treatments in nearly all crops. All crops grown on the nursery yard, and cabbage grown under the high tunnel, had greater yields when grown in medium treatment 3, compared with the IS and C media treatments (P ≤ 0.05). All crops grown in medium treatment 2 on the nursery yard produced greater yields than the IS and C media treatments (P ≤ 0.05).

ARS-Media for Excel: A Spreadsheet Tool for Calculating Media Recipes Based on Ion-Specific Constraints.

ARS-Media for Excel is an ion solution calculator that uses “Microsoft Excel” to generate recipes of salts for complex ion mixtures specified by the user. Generating salt combinations (recipes) that result in pre-specified target ion values is a linear programming problem. Excel’s Solver add-on solves the linear programming equation to generate a recipe. Calculating a mixture of salts to generate exact solutions of complex ionic mixtures is required for at least 2 types of problems– 1) formulating relevant ecological/biological ionic solutions such as those from a specific lake, soil, cell, tissue, or organ and, 2) designing ion confounding-free experiments to determine ion-specific effects where ions are treated as statistical factors. Using ARS-Media for Excel to solve these two problems is illustrated by 1) exactly reconstructing a soil solution representative of a loamy agricultural soil and, 2) constructing an ion-based experiment to determine the effects of substituting Na+ for K+ on the growth of a Valencia sweet orange nonembryogenic cell line.

Effects of macro and micronutrients on neutral lipid accumulation in oleaginous microalgae

ABSTRACTIn this study, effects of key macro and micronutrients on neutral lipid accumulation of six oleaginous microalgae species were investigated. For each nutrient, three different concentrations (0.5×, 1×, and 2×) were tested individually and compared to the most commonly utilized growth medium recipes. Neutral lipid accumulation was quantified using specific nonpolar dyes with long-wavelength absorption and fluorescence. Tested microalgae species showed different lipid accumulation responses to changes in nutrient concentrations. Optimum concentrations of nutrients were selected for each species based on either the most lipid productive case or no significant difference to the lipid contents. Selected optimal conditions were also validated where Chlorella sorokiniana, Chlorella vulgaris, Dunaliella tertiolecta, and Tetraselmis suecica showed 4, 10, 34, and 39% higher lipid productivities per unit volume, respectively. Botryococcus sudeticus and Ettlia oleoabundans did not show improved lipid productivities. Extensive optimization of macro and micronutrient concentrations offers several benefits of decreased material inputs and waste generation, improved biomass productivity, and overall cost savings.

Reviews and Resources:Lab Ref, Volume 1: a Handbook of Recipes, Reagents, and Other Reference Tools for Use at the Bench

As research scientists, some of our most prized possessions are the reference manuals that line our lab bookshelves. Volumes of general and specialized manuals, each with their contents of detailed background information, protocols, reagent and media recipes, reference tables, etc. They are the books we direct students to when they are designing experiments.

Score a tweet and post a goal: Social media recipes for sports stars

Sports performance is not the unique ingredient of a sports star's market value. Some gifted players are undermined by weak media exposure while some less talented players who actively engage in social media and attract fans in millions benefit from exorbitant contracts. This research conceptualizes the effect of social media in the sports business and analyzes the recipes that lead to high market value for sports stars. This study uses qualitative comparative analysis (QCA) on a sample of 95 top soccer players in Europe. The empirical results reveal that sports performance and social media activity are both necessary but insufficient conditions for a high market value condition. This research provides a roadmap for managers and sports stars to navigate in the social media arena.

A simple novel approach for real-time monitoring of sodium gluconate production by on-line physiological parameters in batch fermentation by Aspergillus niger

In this paper, approach for real-time monitoring of sodium gluconate (SG) fermentation was established for the first time by the equations which can calculate real-time key-parameters by on-line physiological data. Based on this approach, limiting factors were found out in initial fermentation F1 and then step-wise agitation increase and improved medium recipe were proposed in fermentation F2 and F3, respectively. The highest average SG production rate (16.58±0.91gL−1h−1) was achieved in fermentation F3, which was 104.2% and 48.0% higher than those in fermentation F1 and F2, respectively. Meanwhile, due to shorter fermentation period (decreased from 34h to 18.7h), lower biomass (about 1.5gL−1) and less by-product accumulation, the overall yield of 0.943±0.012 (molmol−1) in fermentation F3 increased more than 16.0% compared to fermentation F1. This approach had been successfully applied to industrial fermentation and greatly improved SG production.

A Reliable Homemade Tissue Culture Protocol for <i>Dendrobium</i> Orchid Cultivation

Murashige and Skoog (MS) medium, a common tissue culture media for orchids which is mostly supplemented with various natural organic substances with high nutritional values can accelerate the growth of plant tissues. Nevertheless, the knowledge of actual compositions of these natural added substances is limited, causing various culture results. In this study, we have investigated the effects of added natural organic nutrient ratios on 2 orchid tissue cultures, Dendrobium, Dendrobium farmeri Paxt. and Dendrobium griffithianum Lindl. Additionally, simple, affordable medium recipes, MS media supplemented with banana were thus economically sterilized by either a simple type steaming vapor boiler or an autoclave for reliability improvement. The results showed that the orchids grew better in medium supplemented with Namwa banana especially at 150 g/L for Dendrobiumfarmeri Paxt. and 75 g/L for Dendrobium griffithianum Lindl. The suitable steaming procedure at 100°C for 60 minutes was adequate for eliminating most pathogens from the media and helped both orchid seedlings cultivated very well. Moreover, the optimized homemade tissue culture protocol provides fast effectiveness, simplicity, consistently high nutritional values, and practically being suitable technique for ready transferring to the community.

Where to find recipes for media.

Current Protocols in MicrobiologyVolume 36, Issue 1 p. A.2C.1-A.2C.11 UNIT Where to Find Recipes for Media Commonly Used Reagents and Equipment First published: 01 February 2015 https://doi.org/10.1002/9780471729259.mca02cs36Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat View the latest in Microbiology This article also appears in:Equipment and Stock Solutions Appendix RelatedInformation