Effect Of Different Media Research Articles

In vitrogermination and viability of pollen grain of coconut accessions

Storage as a means of maintaining the pollen viability is important for the preservation of the genetic variability, facilitates the exchange of germplasm and greatly contributes to the generation of variability obtained from artificial crosses, increasing the efficiency of breeding programs. The objective of this study was to select different culture media for the in vitro germination of pollen grain of dwarf and tall coconut accessions, as well as to determine the viability of pollen grain at room temperature conditions. For this purpose, Brazil Green Dwarf (BGD) and Brazilian Tall (BRA) pollen grains derived from the Coconut Active Germplasm Bank of Embrapa Coastal Tablelands Sergipe were used. To evaluate the effect of different culture media on the in vitro germination of pollen grains of anão verde do Brasil de Jiqui (AVeBrJ) and gigante do Brasil Praia do Forte (GBrPF) accessions, they were inoculated on to Petri dishes containing 2 ml of culture media. The pollen viability was assessed by staining with 1% acetic carmine and in vitro germination at 0, 24, 48 and 72 hours. The culture medium of Lora is suitable to assess the in vitro germination of pollen grain of the AVeBrJ and GBrPF accessions. The pollen grain of the AVeBrJ accession showed intermediate viability (66.87%) at room temperature up to 23.14 hours by in vitro germination. The pollen grain of the GBrPF accession showed high viability, above 70%, at room temperature up to 120 hours by in vitro germination.

Comparison of the effect of different media on the clinical outcomes of the density-gradient centrifugation/swim-up and swim-up methods.

ObjectiveSperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF.MethodsPatients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123).ResultsThere were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates.ConclusionWe suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

Effect of different media and growth hormones on shoot multiplication of in vitro grown Centella asiatica accessions

Centella asiatica also known as Gotu Kola is an important medicinal plant which contains several bioactive compounds such as triterpenoid saponins including asiaticoside, madecassoside, centelloside, asiatic acid, etc. In addition, Centella sp. contains other components including flavonoids, phytosterols, tannins, amino acids, sugars, etc. Due to its medicinal importance, this plant is being overexploited and it is essential to conserve this plant. In the present investigation, comparative study of different accessions of Centella asiatica for shoot multiplication in different media and growth hormones was performed. Shoot multiplication and phytocompounds production from different accessions is required to be assessed to choose an accession giving optimum production of phytocompounds. For this, different media containing different combinations of macro and micro nutrients have to be tested that influence the growth of the plant. Assessment of best culture media and concentrations of plant hormone for shoot culture are some of the critical culture conditions to achieve. So there is a need to optimise such conditions that will enhance the growth of the plant. For shoot culture, explant of different accession of Centella asiatica were inoculated in different media i.e. Murashige and Skoog (MS), Gamborg’s B5 and Nitsch medium which were supplemented with standard concentrations of plant growth hormones. The cultures were incubated at 25 ± 2°C with photoperiod of 16 hours. After six week of incubation period, highest growth was found in MS media in all accessions. Further MS medium was supplemented with different combinations of growth hormones. After six week of incubation, MS medium supplemented with 1mg/l BAP showed the highest growth of the plant followed by 2mg/l BAP+0.5 NAA.

In Vitro Multiplication of Photinia (Photinia X Fraseri)Usingdifferent Culture Media

A very high rate of multiple shoots was obtained from nodal explants of Photinia x fraseri on MS medium supplemented with a various concentration of Kin alone (1, 2, and 3) mgl-1alone and combination of Kin at (1, 2, and 3) mgl-1 with IBA or IAA at (0.2 and 0.4) mgl-1. The results show that the number of shoots/ explant was significantly affected the levels of Kin alone and combination with IBA or IAA. The number of shoots (6.00 shoot/ explant) when Kin added to the medium at 3mgl-1 in which it was significantly different from other Kin concentrations. While the effect of different medium with the fixed and best concentration of Kin+ IBA and Kin+ IAA on the average shoot number and leaves number /explant of Photinia explant (node segment and shoot tips) The highest number of shoots (5.90) was recorded in node segment when cultured on WPM containing Kin+ IBA and it was increased significantly compared with all treatments from shoot tip. However, the maximum number of leaves (20.10) was recorded on WPM medium supplemented with Kin+ IBA and it was increased significantly compared with all treatments which containing different media plus Kin at 2mgl-1 + 0.4mgl-1 IBA and Kin at 1mgl-1+ 0.4mgl-1 IAA.Shootlel form the combination of from photinia cultured on ¼MS medium improved 0.6 mgl-1NAA concentration produced the highest number of roots/shootlel (6.5roots/shootlet) compared with all treatments contains ½MS medium added IAA, IBA and NAA on all concentration. While, theshootlets from photinia cultured on ¼MS medium and supplemented with 0.6mgl-1NAA formed an average (5.29 cm) compared to all treatments containing ½MS medium. Such plantlets were successfully transferred to soil after hardening with a high rate of survival.

234 NEW CULTURE MEDIA AFFECTS BLASTOCYST DEVELOPMENT AND GENE EXPRESSION LEVELS IN IN VITRO-PRODUCED BOVINE EMBRYOS

The purpose was to examine effects of different media for bovine in vitro oocyte maturation (IVM) and in vitro embryo culture (IVC) on blastocyst rates, morpho-kinetics, and relative abundance of mRNA of 8 genes associated with critical processes and developmental competence in the embryo. Abattoir-derived cumulus-oocyte complexes (COC) were in vitro matured (IVM) in either TCM199 [+0.5% BSA and gonadotropins (Suigonan Vet 150 I.E. mL–1)] or in a novel commercially available media (Bo-IVM), and a total of 1196 presumptive zygotes, from 4 replicates, were submitted to in vitro culture (IVC) and cultured in either SOF (+0.5% BSA) or in a novel commercially available media (Bo-IVC). Blastocyst rates and morpho-kinetics were assessed on Day 8 after fertilization. The high-quality blastocysts from each group were analysed by RT-qPCR, on single blastocysts using earlier verified primers, for BAX, BCL2L1, DNMT3A, FASN, G6PD, HSPA1A, SLC2A1, and SLC2A3. Data on blastocyst rates were analysed for statistical differences using a linear regression model, using a binary reproach and general estimating equations. One-way ANOVA was used to detect differences in the relative abundance of mRNA between groups, whereas differences between maturation and culture media were analysed by a 2-way ANOVA. Blastocyst rates in the Bo-IVM/Bo-IVC (37%), TCM199/Bo-IVC (33%), Bo-IVM/SOF (26%), and TCM199/SOF (28%) groups were significantly different from each other (P < 0.0001). Specifically, the Bo-IVM/Bo-IVC group differed significantly from both SOF-cultured groups (P < 0.01). Subjectively, this group also had embryos of the highest quality and most advanced development. Significantly increased levels of mRNA transcripts were found for embryos cultured in Bo-IVC for all genes (P < 0.05) except BCL2L1. In conclusion, the developmental rates and gene expression of in vitro-produced bovine blastocysts were affected by the use of different culture media. Increased blastocyst rates, apparently superior embryo quality, and more abundant gene expression were achieved when blastocysts were cultured in Bo-IVC culture media compared with SOF.The project was supported by The Danish Council for Strategic Research.

Effects of nutrient media, different cytokinin types and their concentrations on in vitro multiplication of G×N15 (hybrid of almond×peach) vegetative rootstock

Abstract The objective of this research was to assess the effects of different media i.e. Murashige and Skoog (MS) and Quoirin and Lepoivre (QL), cytokinin type i.e. 6-Benzyladenin (BA) and 6-Benzylaminopurine (BAP) and cytokinin concentration on in vitro proliferation of the G × N15 rootstock. To evaluate the effects of different media and cytokinin type, two separate experiments were conducted as factorial based on completely randomized design, and single nodes were used as explants. The results showed that MS nutrient medium was found to be superior to QL nutrient medium. Regarding the interaction between media and growth regulators, the best interaction was found in MS medium supplemented with 1 mg l −1 BAP resulting in 8.5 new micro shoots/explant while 7.75 shoots were observed in MS medium containing 1.25 mg l −1 BA. The longest length of new micro-shoots (2.10 cm) was obtained in hormone-free MS medium. Findings of this study showed that there is a significant correlation between the hormone level and plantlet height and formed callus weight so that an increase in BAP and BA levels in both of MS and QL media resulted significantly in height decrease and callus weight increase. The results also suggest that the best and the worst plantlets in terms of quality were observed in hormone-free QL medium and MS medium supplemented with 1.25 mg l −1 , respectively. These results reflect the fact that the presence of high amounts of NH 4 NO 3 and cytokinin especially BAP in culture medium triggered inhibitory effect on shoot growth.

In vitro development and acclimatization of dendezeiro (Elaeis guineensis)

Fruits and almond from the dendezeiro, oil palmbelonging to the Elaeis genus,are widely used for the production of cookingoils or for the pharmaceutical and cosmetic industries.In the last decade, this oil palm also emerged as a promising source for commercialbiofuel production. This study evaluated the effect of different culture media, MS (MURASHIGUE AND SKOOG) and Y3 (EEUWENS)and carbohydrates duringin vitro germination of zygotic embryos, the effect of growth regulators GA3, NAA and BA Ponin vitro seedling development, and the survival rate of acclimatized seedlingsof Manicoré hybrid (Elaeis oleifera x E. guineensis). Zygotic embryos were inoculated on MS and modified Y3 media, supplemented with different sucrose concentrations (30, 45, and 60 gL-1) or sorbitol (36 gL-1), and the germination rate was evaluated after 30 days. Subsequently, seedlings were transferred to modified Y3 culture medium supplemented with differentGA3 concentrations (3.5 and 7 mgL-1) or without it, combined or not with 1 mgL-1 of NAA, 5 mgL-1 of BAP.The highest germinationpercentage of germinated embryos (92%) was observed in MS medium supplemented with 36 gL-1 sorbitol. Culture media supplemented with growth regulatorsGA3, NAA and BAP promoted greater shoot lengththan control media. Rooted seedlings showed high survival percentage (85%) during acclimatization.

A Study on the Aromatization Reaction of Lanlian FCC Gasoline with Different Medium Gas

By using Lanlian FCC gasoline as a feedstock and LBO-A as a catalyst, the effects of different medium gases on the aromatization product yield, which includes hydrogen or methane and ethane or ethylene or dry gas with steam, were researched in a confined fluidized bed reactor. The experimental result showed that adding hydrogen or methane and ethane or ethylene or dry gas with steam has obvious effects on Lanlian FCC gasoline's yield. Further, the optimizing operation conditions are ratio of hydrogen and hydrogen with steam at about 20 or 40%. Lanlian FCC gasoline and light oil's yields have a maximum 89.24 and 92.36%, respectively.

Efeito de diferentes meios de cultura na germinação in vitro de sementes de mangabeira (Hancornia speciosa Gomes)

A mangabeira, pertencente à família Apocynaceae, é uma espécie nativa do Cerrado e dos tabuleiros costeiros, sendo bastante conhecida pela importância social, econômica e cultural. O objetivo deste trabalho foi avaliar o efeito de diferentes meios de cultura na germinação in vitro de sementes de mangabeira, visando contribuir cientificamente para o conhecimento da espécie. Os tratamentos utilizados foram: T1 - vermiculita (+40 mL de água); T2 - vermiculita + areia (1:1 + 40 mL de água); T3 - vermiculita + areia barrada (1:1 + 40 mL de água); T4 - vermiculita + MS básico (40 ml); T5 - vermiculita + ½ MS (40 ml); T6 - areia (+40 mL de água); T7 - areia barrada (+40 mL de água); e T8 - areia + areia barrada (1:1 + 40 mL de água). Para tanto, realizou-se um experimento com oito tratamentos, incluindo oito repetições com 80 sementes por tratamento. Foram analisadas as seguintes variáveis: porcentagem de germinação, tempo médio de germinação e índice de velocidade de germinação. O delineamento experimental foi inteiramente casualizado e as médias, comparadas pelo teste de Tukey a 5% de significância. Observaram-se diferenças significativas para a porcentagem de germinação, sendo T3 estatisticamente superior, e para o IVG, com o melhor resultado, os tratamentos T1, T2 e T3. Entretanto, os valores de tempo médio de germinação não apresentaram diferença significativa. Diante dos resultados, pôde-se concluir que os tratamentos dotados de vermiculita e combinações, T1, T2 e T3, exercem influência positiva na emergência de Hancornia speciosa Gomes.

Simulation analysis of transformer oil and glycerin as dielectric medium in inductive voltage adders

This paper gives the initial consideration of using glycerin medium to substitute the transformer oil widely used in present cells of magnetically insulated inductive voltage adders (MIVA). The effects of different mediums on MIVA performances such as electric field distribution, feed current uniformity, and cell output voltage, are simulated and compared. The simulation results indicate that, in case of four pulses driving simultaneously, the cells insulated by glycerin would output a pulse with the peak voltage increasing by about 8%. However, in case of four pulses driving separately, as replacing transformer oil with glycerin, the peak voltage of cell outputs decreases by 9%, and risetime increases dramatically from 32 ns to 59 ns. For the encountered issues caused by the glycerin medium, some solution methods are presented, and the feasibility of utilizing glycerin in MIVA cells is discussed.

Optimization of Culture Conditions for Stem Cells Derived from Human Anterior Cruciate Ligament and Bone Marrow

Tissue engineering with stem cells is a fascinating approach for treating anterior cruciate ligament (ACL) injuries. In our previous study, stem cells isolated from the human anterior cruciate ligament were shown to possess extensive proliferation and differentiation capabilities when treated with specific growth factors. However, optimal culture conditions and the usefulness of fetal bovine serum (FBS) as a growth factor in in vitro culture systems are yet to be determined. In this study, we compared the effects of different culture media containing combinations of various concentrations of FBS and the growth factors basic fibroblastic growth factor (bFGF) and transforming growth factor-β1 (TGF-β1) on the proliferation and differentiation of ligament-derived stem cells (LSCs) and bone marrow mesenchymal stem cells (BMSCs). We found that α-MEM plus 10% FBS and bFGF was able to maintain both LSCs and BMSCs in a relatively undifferentiated state but with lower major extracellular matrix (ECM) component gene expression and protein production, which is beneficial for stem cell expansion. However, the differentiation and proliferation potentials of LSCs and BMSCs were increased when cultured in MesenPRO, a commercially available stem cell medium containing 2% FBS. MesenPRO in conjunction with TGF-β1 had the greatest ability to induce the differentiation of BMSCs and LSCs to ligament fibroblasts, which was evidenced by the highest ligamentous ECM gene expression and protein production. These results indicate that culture media and growth factors play a very important role in the success of tissue engineering. With α-MEM plus 10% FBS and bFGF, rapid proliferation of stem cells can be achieved. In this study, MesenPRO was able to promote differentiation of both LSCs and BMSCs to ligament fibroblasts. Differentiation was further increased by TGF-β1. With increasing understanding of the effects of different culture media and growth factors, manipulation of stem cells in the desired direction for ligament tissue engineering can be achieved.

Prediction of the in vivo performance of enteric coated pellets in the fasted state under selected biorelevant dissolution conditions

The purpose of this research was to predict the in vivo dissolution of lansoprazole from enteric coated pellets in the fasted state using a biorelevant flow-through dissolution method with low flow rates and volumes close to those in vivo. Additionally, a novel rotating stirring element, composed from magnet inserted in a silicone tube, was used to produce the movement of the pellets and expose them to slightly increased physical stress. Obtained dissolution results were compared to the dissolution results of our previous work using the USP IV with higher flow rate (11ml/min). As drug release from enteric coated pellets usually starts in the small intestine, the influence of pellets’ residence time in the gastric medium and additionally the effect of different media on drug release was studied. Prolongation of residence time in an acidic medium had only minor effect on the release rate after initial lag time, but significantly reduced the total amount of the drug released from both tested formulations, which was attributed to the drug’s degradation in an acidic medium. The increased physical load on the pellets induced by the rotating stirring element compensated for the decrease of flow rate from 11ml/min using the USP IV to 3ml/min using the non-compendial system. Considering also gastric emptying kinetics good prediction of the in vivo release was achieved compared to in vivo absorption data obtained from a pharmacokinetic study under fasting conditions. Thus, using more physiologically relevant dissolution conditions, expressed through low volume and lower flow rate, and in combination with increased mechanical stress we obtained equally good in vitro/in vivo correlation as using USP IV and higher flow rates. Comparison of the dissolution results obtained with two different systems provided additional insight into product behaviour and improved prediction of in vivo performance.

The impact of media on a new product innovation diffusion: a mathematical model

In this paper, we proposed a three compartment model consisting of non-adopter, adopter and frustrated classes of population to discuss the influence of media coverage in spreading and controlling of adopter of a particular product in a region. The model exhibits two equilibria:(i) a adopter-free and (ii) unique interior equilibrium. Stability analysis of the model shows that the adopter-free equilibrium is always locally asymptotically stable if the influence number of adopter $(R_0)$, which depends on parameters of the system is less than unity. Otherwise if $R_0>1$, a unique interior equilibrium exists, it is locally asymptotically stable under some set of conditions. Further analytically and numerically it is observed that the region for backward bifurcation of adopter population increases with the decrease of the valid contact rate before media alert. Finally, numerically experimentations are presented to establish the effect of different media alert rate on adopter and non adopter population.

Increased Osmolarity and Cell Clustering Preserve Canine Notochordal Cell Phenotype in Culture

Introduction Degeneration of the intervertebral disc (IVD) is associated with a loss of notochordal cells (NCs) from the nucleus pulposus (NP) and their replacement by chondrocyte-like cells. NCs have a stimulatory effect upon other cells, such as native IVD chondrocyte-like cells and mesenchymal stem cells, and therefore they are an interesting focus for regenerative tissue engineering strategies. However, optimal conditions, such as osmolarity and other characteristics of the culture media, for long-term culture of NCs are not known. The purpose of this study was to investigate the effects of different culture media and osmolarity on the physiology of NCs in vitro. Materials and Methods NC clusters isolated from canine IVDs were suspended in alginate beads and cultured at 37°C under normoxic conditions for 28 days. Three different culture conditions were investigated; (1) DMEM/F12 (300 mOsm/L), (2) α-MEM (300 mOsm/L), and (3) α-MEM adjusted to 400 mOsm/L to mimic a hyperosmolar environment. NC morphology, expression of genes related to NC markers, matrix production and remodeling, and DNA- and GAG analyses were performed on 1, 7, 14, and 28 days in culture. Data analysis were performed with the aid of a mixed linear model which incorporated fixed effects (i.e., “time in culture” 1, 7, 14, 21, and 28 days, and NP tissue), “culture condition” (DMEM/F12, α-MEM, and α-MEM 400 mOsm/L), and the interaction between these factors; in the random part of the mixed models, the factor “dog” (dogs 1-6) was incorporated to take the correlation within each dog into account. Benjamini-Hochberg correction was applied to correct for multiple comparisons. Results Large, vesicle-containing cells organized in clusters, characterized as NCs, remained present during 28 days for all culture conditions. However, the proportion of the NC clusters decreased over time, whereas the proportion of spindle-shaped cells increased. When comparing the different culture conditions, on day 28 in culture, the cell population cultured in α-MEM 400 mOsm/L appeared to contain more vacuolated cells (NCs) compared with the DMEM/F12 and α-MEM and hence fewer spindle and polygonal cells (Fig.). Gene expression profiling at 7, 14, and 28 days in culture compared with day 1 indicated an initial loss of NC phenotype followed by recovery of brachyury and aggrecan expression after 28 days of culture supporting a potential recovery of NC phenotype. NCs cultured in α-MEM adjusted to 400 mOsm/L showed the highest expression of brachyury, cytokeratin 18, and aggrecan, the highest GAG production, and the lowest collagen 1α1 gene expression. Conclusion In conclusion, NCs cultured in alginate in native cell clusters under normoxic conditions, partially retained their characteristic morphology and recovered their phenotype in long-term culture. In comparison to using DMEM/F12 (300 mOsm/L) culture medium, the use of α-MEM (300 mOsm/L) culture medium for culturing NCs has a beneficial effect on preserving the NC phenotype in culture, which is further facilitated by increasing the culture medium osmolarity to physiological levels (400 mOsm/L). This may be explained by the difference in composition of DMEM/F12 and α-MEM culture media: α-MEM contains a lower concentration of D-glucose (dextrose) than DMEM/F12 (5.5 vs. 17.5 mM, respectively). Since IVD cells are subject to low glucose concentrations in vivo, the relatively low glucose content in α-MEM may facilitate NCs to retain their phenotype. Moreover, hyperglycemia has been positively correlated with IVD degeneration in diabetic rats and recently it has been shown that a high glucose environment stimulates apoptosis and inhibits proliferation of NCs in vitro. Furthermore, preservation of the NC clusters may facilitate the NC physiology and the preserved extracellular matrix may participate in the presentation of growth factors to the cultured NCs. These findings provide additional information concerning the maintenance of NCs in vitro that may set the stage for further mechanistic inquiry into the biology of NCs and their potential role in designing new regenerative strategies. Acknowledgments This work was supported by AO Spine International, through an AOSpine Reserach Network grant (# 104982). Disclosure of Interest None declared

Increased Osmolarity and Cell Clustering Preserve Canine Notochordal Cell Phenotype in Culture

Degeneration of the intervertebral disc (IVD) is associated with a loss of notochordal cells (NCs) from the nucleus pulposus (NP) and their replacement by chondrocyte-like cells. NCs are known to maintain extracellular matrix quality and stimulate the chondrocyte-like NP cells, making NCs attractive for designing new tissue engineering approaches for IVD regeneration. However, optimal conditions, such as osmolarity and other characteristics of the culture media, for long-term culture of NCs are not known. The purpose of this study was to investigate the effects of different culture media and osmolarity on the physiology of NCs in vitro. NC clusters isolated from canine IVDs were suspended in alginate beads and cultured at 37°C under normoxic conditions for 28 days. Three different culture conditions were investigated; (1) Dulbecco's modified Eagle's medium (DMEM)/F12 (300 mOsm/L), (2) α-MEM (300 mOsm/L), and (3) α-MEM adjusted to 400 mOsm/L to mimic a hyperosmolar environment. NC morphology, expression of genes related to NC markers, matrix production and remodeling, and DNA- and glycosaminoglycan (GAG) analyses were performed on 1, 7, 14, and 28 days in culture. Large, vesicle-containing cells organized in clusters, characterized as NCs, remained present during 28 days for all culture conditions. However, the proportion of the NC clusters decreased over time, whereas the proportion of spindle-shaped cells increased. Gene expression profiling at 7, 14, and 28 days in culture compared to day 1 indicated a initial loss of NC phenotype followed by some recovery of brachyury and aggrecan gene expression after 28 days of culture supporting a potential recovery of NC phenotype. NCs cultured in α-MEM adjusted to 400 mOsm/L showed the highest gene expression of brachyury, cytokeratin 18, and aggrecan, the highest GAG production, and the lowest collagen 1α1 gene expression. In conclusion, NCs cultured in alginate in native cell clusters, partially retained their characteristic morphology and recovered their phenotype in long-term culture. The type of culture medium and medium osmolarity appear to be important factors for culturing NC clusters. These findings provide additional information concerning the maintenance of NCs in vitro that may aid further mechanistic inquiry into the biology of NCs.

Effect of different media on exopolysaccharide and biomass production by the green microalga Botryococcus braunii

Botryococcus braunii is a colonial green microalga with recognized potential to synthesize lipids and hydrocarbons for biofuel production. Besides this ability, this microalga also produces exopolysaccharides (EPS). Nevertheless, there are few reports about their biotechnological aspects and industrial applications. In this study, the effect of the nutritional conditions was examined by using two different culture media (BG11 and D medium). To our knowledge, the latter has not been reported before for culturing B. braunii. After 49 days of incubation, the final production of EPS was found to be statistically higher (P < 0.05) in the D medium (0.549 ± 0.044 g L−1) than in BG11 (0.336 ± 0.009 g L−1). On the contrary, the biomass production was found to be higher in BG11 (1.019 ± 0.051 g L−1) than in the D medium (0.953 ± 0.056 g L−1). However, this difference was not statistically significant. The difference in salinity and nitrogen concentration between both media is suggested as the main factor involved in the EPS and biomass results. FTIR spectra of B. braunii EPS from both media revealed presence of uronic acids and absence of amino and sulfate groups. Despite the similarity between both spectra, there were some different signals (at 1,921.52 and 720.60 cm−1) which may mean a difference in glycosyl composition.

Optimization of a biolistic transformation system for transfer of antifreeze gene KN2 and the bar herbicide resistance gene in common wheat.

We studied the effects of different media for callus induction and differentiation, and pre-culture period of immature wheat embryo culture on biolistic transformation efficiency for including antifreeze gene KN2 and bar conferring resistance to the herbicide bialaphos. The percentage of plantlets generated from induction and differentiation media without Cu2+ was lower than those cultured on differentiation media with Cu2+ (71.15%) or induction media with Cu2+ (68.45%) and both induction and differentiation media with Cu2+ (52.17%). The combinations of Nor medium for callus induction and Cu2+ medium for regeneration, and Cu2+ medium for induction and R medium for regeneration were superior for biolistic transformation. The calli induced on Cu2+ medium and pre-cultured for 4 d before biolistic transformation, and cultured on R medium after biolistic transformation produced the highest percentage (65%) of transgenic plantlets with the KN2 gene. Overall, about 50% plantlets regenerated from calli pre-cultured 4d before bombardment carried the KN2 gene; 44.7% of the plantlets carried the bar gene, which was higher than for any other treatment, followed by pre-culture 1d with 31.43% transformation rate for the KN2 gene and 20% transformation rate for the bar gene.

배양조건에 따른 붉은자루동충하초 균주 간의 균사생장 특성 재평가

붉은자루동충하초의 이용성을 증대하기 위한 기초자료로서 확보하고자 배지종류, 배양법, 산도, 탄소원, 그리고 질소원이 균사생장에 미치는 영향을 4개 균주를 대상으로 재평가하였다. 시험한 8종의 고체배지에서의 균사생장은 배지 종류 및 배양일수에 따라 균주간에 차이가 있었다. 8종의 고체배지 중 PDA 및 MMMA(mushroom minimal medium agar)서 균사생장이 우수하였다. 반면에 8종의 액체배지 중에서는 4개 균주 모두 SDYM(Sabourand's dextrose yeast extract medium) 배지에서 모두 우수한 균사생장을 보였다. 액체배지에서의 균사생장은 pH 4.0에서 pH 9.5 사이의 범위에서 큰 차이 없이 양호한 생장을 보였다. 일부 균주는 pH 8.0에서 pH 9.5의 알카리 조건에서 생장이 양호하였다. 탄소원의 경우 xylose 이용 능력이 균주간에 차이가 크게 나타났다. 질소원 경우에 Urea 이용 능력이 있는 균주가 존재함이 새로이 확인되었다. 이런 결과는 기존에 보고되지 않은 붉은자루동충하초의 새로운 균사생육 특성을 보여준다. 결론으로서 향후 새로운 붉은자루동충하초 균주를 이용하고자 할 경우는 반듯이 사전에 적정 균사생육 조건을 검토하는 것이 필요하다고 사료된다. To obtain basic data for the better use of Cordyceps pruinosa we reassessed the effect of different medium, culture method, pH, and carbon and nitrogen sources on the mycelial growth properties of four C. pruinosa isolates. The growth of mycelia differed among the four isolates depending on medium type and cultivation days. Among the tested 8 kinds of solid media, the four isolates grew well on PDA and MMMA(mushroom minimal medium agar). While, among the tested 8 kinds of liquid media, all the isolates grew well in SDYM(Sabourand's dextrose yeast extract medium). The isolates also grew well in the SDYM with pH from 4.0 to 9.5 without any inhibition. One isolate could best grow at pH 8 to 9.5. Regarding the ability of utilizing carbon source, the difference of mycelia growth among the isolates was the most with xylose. Regarding nitrogen source, three isolates could utilize urea which is new fact in this species. These results provide new points on the growth properties of the fungal mycelium which has not been explored before. Overall, this reassessed study concluded that it is necessary to check in advance the growth properties of mycelium when a new isolate of C. pruinosa is expected to be used for application.

Study on the Effects of Different Media on the Seeding of F. cirrhosa at Plateau

Bulbus Fritillaria cirrhosa is a precious wild herb of Liliaceae Fritillaria which grows in Tibetan area of China. Ripe seeds and one- year bulbs of Fritillaria cirrhosa D. Don were planting materials. Sandy soils, humus soils and humus soils with the dung of Hepialidae Larvae (Lepidoptera, Hepialidae) were selected as nursery materials.The experiments were conducted at XinduqiaoTown of Kangding County (Latitude: 101°, 34'N, longitude: 30°, 03'E, altitude: 3500 m) from March to Augustin 2011. The research was aimed to select the optimum medium of productive seeding and to study on effects of different media on growth of F. cirrhosa seeding. The results were: (1) Sowing with seeds and one-year-old bulbs of F. cirrhosa, S1 (100% humus soils with dung of Hepialidae larvae) was the best nursery media for sprouting. (2) The integrated performance was considered, including transverse diameter, longitudinal diameter, the ratio of longitudinal diameter to transverse diameter, 30-grain fresh weight and 30-grain dry weight. Sowing with F. cirrhosa seeds, S1 (100% humus soils with dung of Hepialidae Laravae) was the best medium for promoting the seedlings and getting larger size of bulbs. Sowing with one-year-old F. cirrhosa bulbs, S1 (100% humus soils with dung of Hepialidae Laravae) and S2 (100% humus soils) were advantageous for the growth of plantlet bulbing and getting larger size of bulbs. Adding sandy soil to substrates or 100% sandy soil asnursery media could inhibit thegrowth of F. cirrhosa bulbs, getting better shape of F. cirrhosa bulbs.

Evaluation of the effects of different culture media on the myogenic differentiation potential of adipose tissue- or bone marrow-derived human mesenchymal stem cells

The creation of functional muscles/muscle tissue from human stem cells is a major goal of skeletal muscle tissue engineering. Mesenchymal stem cells (MSCs) from fat/adipose tissue (AT-MSCs), as well as bone marrow (BM-MSCs) have been shown to bear myogenic potential, which makes them candidate stem cells for skeletal muscle tissue engineering applications. The aim of this study was to analyse the myogenic differentiation potential of human AT-MSCs and BM-MSCs cultured in six different cell culture media containing different mixtures of growth factors. The following cell culture media were used in our experiments: mesenchymal stem cell growth medium (MSCGM)™ as growth medium, MSCGM + 5-azacytidine (5-Aza), skeletal muscle myoblast cell growth medium (SkGM)-2 BulletKit™, and 5, 30 and 50% conditioned cell culture media, i.e., supernatant of human satellite cell cultures after three days in cell culture mixed with MSCGM. Following the incubation of human AT-MSCs or BM-MSCs for 0, 4, 8, 11, 16 or 21 days with each of the cell culture media, cell proliferation was measured using the alamarBlue® assay. Myogenic differentiation was evaluated by quantitative gene expression analyses, using quantitative RT-PCR (qRT-PCR) and immunocytochemical staining (ICC), using well-defined skeletal markers, such as desmin (DES), myogenic factor 5 (MYF5), myosin, heavy chain 8, skeletal muscle, perinatal (MYH8), myosin, heavy chain 1, skeletal muscle, adult (MYH1) and skeletal muscle actin-α1 (ACTA1). The highest proliferation rates were observed in the AT-MSCs and BM-MSCs cultured with SkGM-2 BulletKit medium. The average proliferation rate was higher in the AT-MSCs than in the BM-MSCs, taking all six culture media into account. qRT-PCR revealed the expression levels of the myogenic markers, ACTA1, MYH1 and MYH8, in the AT-MSC cell cultures, but not in the BM-MSC cultures. The muscle-specific intermediate filament, DES, was only detected (by ICC) in the AT-MSCs, but not in the BM-MSCs. The strongest DES expression was observed using the 30% conditioned cell culture medium. The detection of myogenic markers using different cell culture media as stimuli was only achieved in the AT-MSCs, but not in the BM-MSCs. The strongest myogenic differentiation, in terms of the markers examined, was induced by the 30% conditioned cell culture medium.