similis-venom serum or pre-immune serum in a shaved region of the

similis-venom serum or pre-immune serum in a shaved region of the back. PBS was used as a negative control. Animals were then euthanized at 2, 4, and 8 h post-injection, and skin samples were taken for histological studies. Rabbit skin samples

were fixed in 10% buffered formalin solution, pH 7.4, and then embedded in paraffin. Sections of 4 μm thickness were stained with hematoxylin-eosin (H&E), Masson Trichrome (with aniline blue, Easypath Special Stain Kit, Brazil) and Reticulin (Easypath Special Stain Kit, Brazil) according to the manufacturer’s instructions. Fixed tissue samples were evaluated by light microscopy. Qualitative microscopic analysis evaluated the presence of necrosis, edema, hyperemia, hemorrhage, and thrombosis. Moreover, inflammatory cell infiltrate was quantitatively assessed using the following approach: a cell count was performed in 50 random fields and the standard deviation BIBW2992 cell line was calculated using 10 samples according to Moro et al. (2003). We determined that the coefficient of variation was stabilized after counting cells in 30 fields (data not shown). Therefore, the total number of inflammatory cells was determined in 30 fields

per slide using a digital image analyser. Morphometry was performed using the specific software Image-Pro Plus 5.0 (Media Cybernetics, MD, USA). The two-way analysis of variance (two-way ANOVA) with Bonferroni post hoc test was used to compare the titration curves of rabbit find more sera. For the in vitro neutralization assay, the one-way ANOVA with Dunnett’s Multiple Comparison Cediranib (AZD2171) post hoc test was used to compare the sphingomyelinase activity of the venom with antivenom group versus the venom only group. For morphometry, the two-way ANOVA with Bonferroni post hoc test was used to test how the inflammatory cell infiltrate count was affected by the groups (venom or serum) or time (2, 4, or 8 h). The level of significance was set at p < 0.05 and statistical analysis was performed using GraphPad

Prism version 5.0 for Windows (GraphPad Software, CA, USA). The immunization protocol was performed in rabbits by injecting several boosters of L. similis venom. We observed that after the third booster, no statistically significant difference was found between the boosters given to the same animal (data not shown). The titration of rabbit sera generated against L. similis venom is shown in Fig. 1. Antivenom antibodies strongly reacted against L. similis venom extracted from male and female spiders ( Fig. 1A and B, respectively). No significant differences were observed between reactions of anti-L. similis and anti-L. intermedia venom sera. The absorbance values of anti-L. similis-venom serum against L. similis male venom ( Fig. 1A) were not statistically different from those obtained for anti-L. similis-venom serum against L. similis female venom ( Fig. 1B). The neutralization assay was evaluated in rabbits immunized with L. similis venom extracted from female spiders ( Fig.

20 was fitted and the result was similar to the result in Table 3

20 was fitted and the result was similar to the result in Table 3 (not shown). A linear predictor based on LKB1 CN,

KRAS mutation, N stage and diagnosis age was constructed from the previously described multivariate model by using the model estimates. ROC curve was used to evaluate the prediction of this multivariate model ( Fig. 3). Area under the curve (AUC) was estimated to be 0.832 and significantly different from 0.5 (p < 0.001, CI: 76.6–93.5%). The ROC curve suggests the test performance over a range of potential cut points on this linear predictor. The optimal cut points cannot be clearly defined because this will depend on user preference for defining false positives and false negatives. For example, with a false positive rate of approximately 30%, the model successfully captured APO866 cell line 80% of the true brain metastasis patients. The poor outcomes of patients with lung

cancer have been widely reported, including the frequent occurrence of brain metastases in patients who have otherwise controlled their disease through primary therapy. In small cell lung cancer http://www.selleckchem.com/products/AZD0530.html progress has been made toward preventing brain metastases through prophylactic cranial radiation which has a proven survival benefit [21]. Attempts to extend this benefit to patients with NSCLC have similarly documented a small benefit in terms of prevention of brain metastasis but without impact on survival [22] and [23]. Such data suggest that biomarkers to enrich for patients at highest risk for brain metastasis may be necessary to make progress in NSCLC. LKB1 is one of the most important tumor suppressor genes and is observed to be inactivated in approximately 30% of all NSCLCs [8]. LKB1 encodes a widely expressed serine/threonine protein kinase whose primary action is through 5′-AMP-activated protein kinase (AMPK) to regulate metabolism and ensure efficient energy production in times of stress [24]. Decreased expression of AMPK

pathway genes have also been shown to be related to metastasis in NSCLC [25]. AMPK controls cell proliferation through the mammalian target of rapamycin (mTOR) kinase, which regulates numerous downstream targets [26]. LKB1 loss impairs downstream AMPK signaling, leading to unsuppressed cell proliferation. LKB1 deficiency can be associated Pyruvate dehydrogenase with increased expression of genes believed to control angiogenesis and metastatic potential [9]. LKB1 can be inactivated through a variety of mechanisms, including gene mutation, deletion and epigenetic events, like promoter methylation. Somatic mutations, mainly nonsense or frameshift mutation, can result in truncated and dysfunctional proteins [27]. As has been shown in this study as well as previous research [5] and [28], somatic mutation can account for only a small fraction of tumors and cannot be the sole reason of LKB1 inactivation.

The development of the resistance

can be clonal/thus not

The development of the resistance

can be clonal/thus not present at all the tumour sites, supporting a concept of continuing the targeted treatment even beyond tumour progression. Co-targeting molecular pathways such as P13K-AKT and/or RAS-ERK and/or T790M or c-Met along with ErbB receptors may result in more optimal anti-cancer effects. We need to better understand the interplay between various oncogenes and tumour suppressors and thus identify key molecular pathways for selleck screening library the treatments. Understanding the reasons for toxicities of targeted therapies will be important for our future rational approaches in combining or sequencing different targeted agents. Co-targeting receptors and their ligand synthesis might help eliminating more effectively receptor activation and downstream oncogenic signalling. New insights of autocrine activation of receptors might lead to new therapeutic approaches. The past successes and failures of therapies led to development of new generation irreversible ErbB family inhibitors and the discovery of new targets, i.e. EML4–ALK fusion gene, ROS, RET and others, which offer significant improvements in clinical outcome for a specific group of patients. The combined regimen strategies of first generation ErbB family inhibitors with anti c-MET inhibitors MK 1775 are being tested in ongoing clinical trials in hope to further improve therapeutic effect. We have to target

multiple pivotal players of malignant cells on individual basis and in each line of treatment, in order to replace “chemotherapy to fit all” by personalized medicine and thus conquer NSCLC. “
“Takashi Yoshimura received his BS and PhD from Nagoya University. Currently, he is a Professor of Animal Physiology and runs three laboratories:

two laboratories at Nagoya University, in the Graduate School of Bioagricultural Sciences and the Institute of Transformative Bio-Molecules (WPI-ITbM), and another at the National Institute for Basic Biology (NIBB) in Okazaki. In the laboratory at the Graduate triclocarban School of Bioagricultural Sciences, he studies the underlying mechanisms of vertebrate seasonal reproduction and circadian rhythms using organisms such as tunicates, fish, birds, and mammals. Based on the findings in this laboratory, he is collaborating with cutting-edge synthetic chemists and theoreticians at WPI-ITbM to develop ‘transformative bio-molecules’ that will improve animal production and human health. The NIBB is one of the host institutes for medaka bioresources of the National BioResource Project of Japan, and provides an excellent opportunity to study medaka fish as a model for seasonal biology. Dr Yoshimura is now studying the underlying mechanism of seasonal time measurement using medaka collected from a range of sites across Japan, because medaka from different latitudes exhibit different seasonal responses.

There is controversy in discussions about this procedure because

There is controversy in discussions about this procedure because many physicians report that the complication rate is almost the same as without a filter. Fig. S10 (online supplementary file) demonstrates that particles captured in the filter can escape if the closing of the filter occurs during the diastolic phase. It has been demonstrated that it is crucial that the

filter is closed during the systolic phase to prevent this escape. From these experiments the following can be clearly seen: All three velocity components have to be measured. The flow rate ratio between the internal and external carotid artery is the most important and significantly influences the flow separation region. The experiments show that particles in flow separation regions sometimes Dabrafenib rotate over several pulse cycles before they are washed away. They can, however, suddenly adhere to the wall and remain there. The Omipalisib solubility dmso pulse wave is not strong enough to wash these small deposits away. The procedure of plaque formation starts. More particles are attracted to and adhere to this area and the flow rate ratio is altered because of the higher resistance caused by the deposits. This effect continues and the stenosis enlarges. The geometry only plays a significant role in these regions with larger bifurcation angles, >40°, where a backward flow is created. In 3D measurements, the calculated shear stresses are

up to 20% higher than those found when measuring only the axial velocity component. With an increasing flow rate, the separation region is slightly reduced but the shear stresses increase. 10–16 Pa are the highest shear 3-oxoacyl-(acyl-carrier-protein) reductase stresses in a healthy carotid artery and are found just at the apex. Shear stresses higher than 180–250 Pa have been measured in models with 90% stenosis for 100–200 ms. Downstream of such stenoses, vortices are created where particles can remain over several pulse cycles. They can also adhere to the wall, creating a growing stenosis. Oscillation

causes shear stresses between 1–40 Pa in such recirculation zones. Biochemical reactions are released. It is very important that stents have to be placed precisely. End threads or wires should never reach into the vessel lumen. Filters have to be closed during the systolic phase, so that no particles escape during the diastolic phase, before they can be pulled out. Experimental studies including MRI, ultrasound measurements and new ultrasound imaging which can measure all three velocity components will be increasingly important in the future to aide in training and refinement of diagnostic and therapeutic procedures. “
“Wall shear stress (WSS), the friction force of flowing blood that acts on the endothelial wall, can vary considerably throughout the vascular beds and has shown to be altered at the outlet or at the inner curvature of arteries, respectively. In an animal model, Cheng et al.

2 have less basic amino acids residues in the C-terminal region w

2 have less basic amino acids residues in the C-terminal region when compared with Kv1.3 high affinity toxins. Such statements could be confirmed in the current work, since Ts15, which has 7 basic residues in its primary structure (Fig. 2) and only 1 in the C-terminal region, shows Selleckchem Olaparib a higher blocking effect to Kv1.2 isoform. Since the amino acid sequence of Ts15 shows a low similarity with that of other toxins, the presence of a functional dyad could not be determined by molecular modeling. To this end NMR or crystallographic studies will be essential. Extensive studies have shown an increasing interest for highly specific blockers of Kv1.3 channels. Since this isoform plays an important

role in the regulation of membrane potential and calcium signaling in lymphocytes cells, it can be used as a therapeutic target for immunosuppressants (Gutman et al., 2005 and Beeton et al., 2006). On the other hand, the

therapeutic application of Kv1.2 blockers is not well elucidated, in view of the fact that this subtype is widespread in the central nervous system and is also able to BAY 80-6946 price form heterotetramer channels (Coleman et al., 1999 and Corzo et al., 2008). It is assumed that this subtype is responsible for maintaining the membrane potential and modulation of electrical excitability in neurons and muscle, however the pharmacological properties can vary between heterotretameric and homotetrameric channels (Coleman et al., 1999 and Gutman et al., 2005). In the present study, we have reported Chlormezanone that Ts15 is capable of blocking both Kv1.2 and Kv1.3 channels with a higher efficiency for the Kv1.2 isoform (Fig. 3 and Fig. 4). Ts15 can be a potential model for the development of new therapeutic drugs. The significant differences in affinity and blocking efficiency observed,

not only between Kv1.2 and Kv1.3, but among all isoforms tested, can be useful to establish critical residues of channel/toxin interaction and therefore help to design a highly specific ligand for a particular channel subtype. Additionally, the low primary structure similarity found between Ts15 and the known KTxs, justifying its classification into a new subfamily, may unveil the existence of other unknown regions and/or important residues for the toxin/channel interaction. The poor specific ligand/channel binding can result in adverse side effects. For instance, Kaliotoxin 1 inhibits Kv1.3 in the process to suppress T cell activity, but is also capable to block Kv1.1 with a potency enough to produce undesirable side effects, such as diarrhea (Crest et al., 1992, Vianna-Jorge et al., 2003 and Beeton et al., 2006). Recently, Takacs et al. (2009), reported the design of a specific ligand able to inhibit Kv1.3 without increasing gastrointestinal mobility due to off–target interactions with Kv1.1. Those studies highlight the importance to define the critical residues for toxin/channel interaction and therefore provide information to design new therapeutic drugs.

Uiboupin & Laanemets (2009) showed that as much as 40% of the are

Uiboupin & Laanemets (2009) showed that as much as 40% of the area of the Gulf of Finland can be under the influence of upwelling during extreme conditions. For example, in 2006 the cross-shore extent of upwelling in the Gulf of Finland was 25 km (1/3 of the width of the Gulf of Finland) and the alongshore extension was 360 km (Suursaar & Aps 2007). On the Polish coast upwelling has Quizartinib mw most often been found to take place off the Hel Peninsula in the Gulf of Gdańsk (see e.g. Matciak et al., 2001 and Myrberg et al., 2010). The potential maximum area of all upwelling on the Polish coast is 10 000 km2, which is ca 30% of

the Polish economic zone (Krężel et al. 2005). Statistical studies of upwelling have been carried out before. Myrberg & Andrejev (2003) determined an upwelling index based on the numerical calculation of vertical velocity

for a 10-year period (1979–1988). A similar study was carried out by Kowalewski & Ostrowski (2005) based on a 7-year experiment of calculated vertical velocities in the southern Baltic. The present paper extends the statistical investigation of Baltic Sea upwelling events based on the integrated use of observations and modelling to cover – for the first time – the entire sea area. For the years 1990–2009, weekly sea surface temperature (SST) maps based on NOAA/AVHRR satellite data were used to evaluate the properties of upwelling during the thermally stratified period from May to Casein kinase 1 September, that buy Fulvestrant is to say, when upwelling is strong enough to raise the thermocline to the surface, thus producing an SST signal. To obtain an independent estimate, numerically simulated daily averaged SST maps were analysed for the same period. Furthermore, favourable and unfavourable wind conditions for upwelling were determined from the wind forcing used as model input. The structure of the paper is as follows: after this introduction, data and methods

are briefly described. Then the results of the statistical analysis are discussed for the period 1990–2009; they are also compared with previous studies. A trend analysis over the total period and for individual months is carried out for identified upwelling areas. Furthermore, for specific upwelling locations, 10-m winds are discriminated into upwelling-favourable and -unfavourable wind conditions, and the relation between upwelling and wind forcing is studied. The paper concludes with a discussion on potential changes in upwelling regions as a consequence of changing climate (wind) conditions. The analysis of upwelling regions and their occurrence is based on SST data with a horizontal resolution of about 1 km calculated from NOAA/AVHRR satellite data for the period 1990–2009. The accuracy of the satellite measurement (cloud detection has been carried out) in comparison with in situ data is about 0.5 °C (Siegel et al. 1994).

Under current technological limitations, the US Department of Agr

Under current technological limitations, the US Department of Agriculture (USDA) in collaboration with the University of

California at Berkeley are trying to develop a genetically engineered switchgrass variety that contains up to 250% more starch than conventional varieties [12] and [13]. This would allow for increasing the economic efficiency of sugar yields and minimizing the final switchgrass-based biofuels costs. If combined with the enzymatic modifications as described above, the production costs of cellulosic ethanol could be reduced substantially. Bortezomib supplier Another feedstock to be potentially used for cellulosic ethanol production in the future is elephant grass (napiergrass) (Pennisetum purpureum) that was introduced to the US from Africa in 1913. This tropical plant is fairly drought-tolerant, grows well on marginal lands and filters nutrients out of runoff in riparian areas. In addition, it does not require irrigation and is capable of producing biomass until the first frost. The main technological requirement and challenge to make napiergrass an efficient and competitive feedstock is to

improve its yields and increase disease resistance [14] and [15]. Poplar has been considered for a long time as a viable find more and prospective feedstock for cellulosic ethanol production in the US. Poplar is drought-tolerant and capable of growing on marginal lands. If indeed grown on abundant or marginal land, it does not compete with other crops for food and animal feed. If cultivated on a biofuel farm, poplar trees create favorable wildlife habitats and provide recreational services. By removing

contaminants from soil, poplar has a valuable potential of soil remediation (phytoremediation) [16], which clearly benefits other parts of the ecosystem chain. Growing poplar trees is said to be more fuel efficient and generates a lower carbon footprint nearly than other annual food crops. Its growth rate is considerably slower than that of biofuels oil crops (e.g., crambe and camelina) [17]. However, this problem could be mitigated by applying biochemical modifications, as discussed in the previous section, or nocturnal photosynthesis that allows poplar to absorb carbon dioxide also at night. This feature allows the plants to reach a higher growth rate with lower water requirements (8–16 inches = 203–406 mm) of precipitation annually) as compared with traditional biofuel crops that require 20–40 inches/year (508–1,016 mm/year) [17]. Another possibility to increase poplar growth rates, which also constitutes a major challenge nowadays, is growing genetically modified poplar species that would hold the nocturnal photosynthesis mechanism and thus constitute a feedstock even more tolerant to drought than the conventional poplar species [18]. One of the possible limitations could be harvesting, transport and storage costs. Another feedstock theoretically considered for ethanol production is orange (citrus fruit) peels. Global agriculture produces about 15.

The most important components of the sprat’s diet are micro- and

The most important components of the sprat’s diet are micro- and mesozooplankton – copepods, cladocerans and rotifers. The diet of the herring is dominated by micro- and mesozooplankton in the first period of life, but older fish consume mainly mysidaceans (macrozooplankton) (Załachowski et al., 1975 and Wiktor, Selisistat chemical structure 1990). The copepods in the sprat and herring diet are represented mostly by Pseudocalanus minutus elongatus, Acartia spp. and Temora longicornis ( Załachowski et al., 1975 and Wiktor, 1990). Copepods are the most abundant zooplankton species

in the Baltic Sea and adjacent waters. Numerous environmental factors – most importantly, temperature – govern essential physiological and metabolic processes in copepods. Together with food quality and concentration, this affects mortality CHIR99021 rates (Hirst & Kiørbe 2002), egg production (Halsband-Lenk et al. 2002) and the growth and development rates of these animals (Twombly and Burns, 1996, Campbell et al., 2001, Peterson, 2001, Hirst and Kiørbe, 2002, Leandro et al., 2006a and Leandro

et al., 2006b). In copepods, stage durations decrease and growth rates increase significantly with temperature, causing the animals to develop faster (Leandro et al., 2006a and Leandro et al., 2006b). Temperature also has a very important influence on moulting rates in juveniles (Hirst & Bunker 2003). Experiments on the growth rate of T. longicornis suggest that this parameter is directly proportional to food concentration ( Harris and Paffenhöfer, 1976a, Harris and Paffenhöfer, 1976b and Klein Breteler et al., 1982) and is strongly influenced

by food quality ( Klein Breteler et al. 1990). The development of T. longicornis has also been found to accelerate with temperature ( McLaren, 1978, Martens, 1980, Klein Breteler and Gonzalez, 1986, Hay et al., 1988 and Fransz et al., 1989). However, the combined effect of food concentration and temperature as a function of these parameters on the growth and development rates of T. longicornis at each of the model stages (naupliar, C1, C2, C3, C4, C5) is Phosphoglycerate kinase established in this paper. Recently, quantitative expressions describing the effects of temperature and food concentration on the growth and development of P. minutus elongatus and Acartia spp. were presented by Dzierzbicka-Głowacka, 2004, Dzierzbicka-Głowacka, 2005a and Dzierzbicka-Głowacka, 2005b) and Dzierzbicka-Głowacka et al., 2006 and Dzierzbicka-Głowacka et al., 2009a. The experimental data given by Klein Breteler and Gonzalez, 1986, Klein Breteler et al., 1982 and Klein Breteler et al., 1990were sufficient to do likewise for T. longicornis. The present work advances the idea of establishing the combined effect of temperature and food concentration on the development and growth of the naupliar stage and copepodid stages (C1, C2, C3, C4, C5) of T. longicornis.

These observations are in good agreement with data reported by De

These observations are in good agreement with data reported by Depasquale and Thompson [6] which demonstrated that PAR-1 expression is a negative prognostic factor in melanomas and strongly correlates with tumor stage. Patients with B-CLL, in most cases, have an indolent clinical course which is asymptomatic and requires no treatment [19]. On the other hand, B-ALL is believed to derive from blockade in maturation of bone marrow lymphoid progenitors leading to bone marrow infiltration, occurrence of various cytopenias in peripheral blood and appearance of blast cells with high proliferative ability. Therefore, patients with B-ALL show

a more aggressive clinical BTK inhibitor research buy behavior than patients with B-CLL [20]. In this study we observed that patients with B-CLL showed similar PAR-1 expression levels in comparison to lymphocytes from healthy individuals. On the other hand, patients with B-ALL exhibited, on average, a significant increase in the expression of this receptor when compared to normal lymphocytes. Interestingly, patients classified as at high risk showed the highest PAR-1 levels among B-ALL patients. CML is a myeloproliferative disease which is characterized by the presence of the fusion gene BCR-ABL, an oncoprotein generated by reciprocal translocation between chromosomes 9 and 22, t(9;22) [21]. In this study,

flow cytometric analyses demonstrated that CML patients in the chronic phase (CML-CP) exhibited a significant decrease in PAR-1 when compared to

granulocytes from healthy individuals. Since PAR-1 has been implicated in inflammatory GSK269962 clinical trial responses as well as in innate and adaptive immunity [22], it is possible PLEK2 that down-regulation of this receptor may contribute for reduced function of these cells and increased susceptibility to recurrent infections in CML. Progression of CML-CP to CML-BP is not fully understood. In fact it is believed that BCR-ABL in cooperation with other factors may account for accelerated leukemogenesis and drug resistance in the acute phase [21]. In this study, we identified an increased PAR-1 expression in blasts from CML-BP patients. However, it is clear that a more extensive analysis is needed to determine the biological significance of these results. Acute myelomonocytic leukemia, which comprises subtypes M4 and M5, are highly aggressive and have a median survival time of only 12 months [23]. Samples from AML-M4/M5 patients that were analyzed in this study exhibited high levels of PAR-1 receptor when compared to monocytes and granulocytes from healthy donors. In contrast, patients with AML-M3 showed PAR-1 expression levels that were similar to those found in granulocytes from healthy donors. However, 3 out of 10 patients exhibited high levels of this receptor.

1A and B) The recovery values for the short-term stored samples

1A and B). The recovery values for the short-term stored samples were lowest for the protein-free cryomedium with 5% DMSO (69.00 ± 6.66%) and highest using BSA (77.40 ± 4.97%). Directly after thawing of the PBMC after short-term storage, the cell viability was high in all samples with values over 96%, independent of the cryomedium. selleck inhibitor Viability decreased marginally in all samples after overnight rest (Fig. 1C) with results between 91.05 ± 1.90% (protein-free medium with 5% DMSO) and 94.75 ± 1.59% (FBS with 10% DMSO). After long-term storage, no significant differences in the recovery and viability could be detected (Fig. 1B and D), compared to the short-term results. The recovery results, normalized to

the short-term performance, show a mean value of 1.00 ± 0.05 directly after thawing and 0.97 ± 0.05 24 h later. Additionally, the viability of all samples, both directly after thawing (1.00 ± 0.00) and after overnight rest (1.01 ± 0.01), was identical to the short-term results. Cryopreservation in all serum-free cryomedia gave high viability and recovery values with maximum results for both the BSA-based medium and the protein-free medium with 10% DMSO. Cells were long-term stable in all of the cryomedia. The maintenance of T-cell responses during cryopreservation is most important. Therefore, PBMC cryopreserved in the five different media were tested in IFN-γ BMS-907351 research buy ELISpot using CMV

and CEF peptide pools as immunogenic antigens after up to 4 weeks and after, on the average, 6 months of storage (Table 1). To classify positive responses, the average number of spot forming cells (SFC) per 106 PBMC was determined; three replicates were used for this calculation. Following “Standardization and Validation Issues of ELISpot Assay” (Janetzki et al., 2005) we used the definition of responder R > 4D and R > 55 [R is the reagent SFC/106 PBMC, D corresponding Dichloromethane dehalogenase to SFC for diluent (background)]. Using the definition above, after short- and long-term storage 12/13 PBMC samples were responsive to the CMV and 9/13 to the CEF (Table 1) peptide pool,

independent of the cryomedium. Also, with 0 to 12 spot-forming cells per 106 PBMC, the background was very low, independent of cryomedium used and storage duration (data not shown). The results indicated, that after cryopreservation using the HSA-based medium, the specific reactivity mainly against CMV antigens was reduced in several samples (e.g. donors #1 and #5, Table 1), whereas using the protein-free medium with only 5% DMSO, the response against both stimulants was decreased, independent of storage duration (e.g. donors #6 and #12, Table 1). In contrast, the response to antigen stimulation after cryopreservation using BSA-based medium and protein-free medium supplemented with 10% DMSO was comparable to the FBS-based medium. In summary, these two media represent alternatives to serum-containing media.