Comparative Analysis of Mesenchymal Stem Cells Cultivated in Serum Free Media

Stem cells are attractive candidates for the regeneration of tissue and organ. Mesenchymal stem cells (MSCs) have been extensively investigated for their potential applications in regenerative medicine and cell therapy. For developing effective stem cell therapy, the mass production of consistent quality cells is required. The cell culture medium is the most critical aspect of the mass production of qualified stem cells. Classically, fetal bovine serum (FBS) has been used as a culture supplement for MSCs. Due to the undefined and heterologous composition of animal origin components in FBS, efforts to replace animal-derived components with non-animal-derived substances led to safe serum free media (SFM). Adipose derived mesenchymal stem cells (ADSCs) cultivated in SFM provided a more stable population doubling time (PDT) to later passage and more cells in a shorter time compared to FBS containing media. ADSCs cultivated in SFM had lower cellular senescence, lower immunogenicity, and higher genetic stability than ADSCs cultivated in FBS containing media. Differential expression analysis of mRNAs and proteins showed that the expression of genes related with apoptosis, immune response, and inflammatory response were significantly up-regulated in ADSCs cultivated in FBS containing media. ADSCs cultivated in SFM showed similar therapeutic efficacy in an acute pancreatitis mouse model to ADSCs cultivated in FBS containing media. Consideration of clinical trials, not only pre-clinical trial, suggests that cultivation of MSCs using SFM might offer more safe cell therapeutics as well as repeated administration due to low immunogenicity.

Colistin Dependency among Colistin-Heteroresistant Acinetobacter baumannii Isolates

Colistin dependent (CD) isolates are dependent on colistin for optimal growth. Here we aimed to systematically determine the emergence of CD among colistin-heteroresistant carbapenem-resistant Acinetobacter baumannii (CRAB) isolates. We also examined the phenotypic characteristics of CD and the evolution of CD strains to overt resistance. Additionally, we examined whether detection of growth in blood cultures was impaired by CD. Heteroresistant isolates, as determined by population analysis profiling, were exposed to colistin; when the colony count with colistin was significantly higher than without, isolates were suspected to be CD. CD was confirmed by Etest and growth curves. CD strains with colistin minimum inhibitory concentrations > 2 mg/L after growth in colistin-free media were considered colistin-resistant. Of the 65 heteroresistant strains tested, eight became CD after colistin exposure. These strains attained higher colony counts and growth rates with colistin vs. without, and grew adjacent to the colistin Etest strip. CD strains exhibited increased susceptibilities to multiple antibiotics compared to their parent heteroresistant strains. All CD strains tested became colistin-resistant following growth without colistin. CD strains were detected in blood culture bottles, but time to detection was significantly prolonged compared with parent strains, suggesting that CD may lead to delay in detection of CRAB bacteremia.

Verbesserte Tribokorrosionsbeständigkeit des martensitischen Stahls X54CrMoVN17-1 durch die Erzeugung von expanded martensite

Seal-free, media-lubricated rolling bearings have a high-energy efficiency as the absence of the seal minimizes frictional loss and increases the efficiency of the driven machine. In addition, the environment is protected by the absence of hazardous lubricants. However, media-lubrication increases tribocorrosive attack on the bearing surface. Therefore, the tribocorrosion resistance of the bearing surface can be increased by a thermal surface treatment called low-temperature plasma nitriding. The produced “expanded martensite” in martensitic steels features a high hardness with comparatively good corrosion resistance. Tribocorrosion tests in 0.9 % NaCl-solution show that the material loss could be reduced by 70 % due to expanded martensite compared to the initial state of the steel.

Influence of media and cellular origin on growth factors secreted by mesenchymal stem cells

Objective: Investigating influences of the culture media and cellular origin on the expression of several growth factors carried by exosomes secreted from umbilical cord-derived mesenchymal stem cells (UCMSC) and bone marrow-derived mesenchymal stem cells (BMMSC). Methods: 03 UCMSC samples was cultured in StemMACs and PowerStem, 03 BMMSC samples was cultured in StemMAC sand ACF Plus. At the passage 2, conditioned media were collected for exosome isolation. Exosomes were then investigated for the expression of growth factors, including: vascular endothelial growth factor A (VEGF-A), fibroblast growth factor 2(FGF-2), hepatocellular growth factor (HGF) and platelet-derived growth factor BB(PDGF-BB). Results: Both UCMSC and BMMSC at secreted exosomes carrying growth factors with different amount. UCMSC cultured in PowerStem could secret exosomes with the largest amount of growth factors. Besides, UCMSC and BMMSC cultured in StemMACs secreted exosomes carrying fewer amounts of growth factors. Secretion of growth factors through exosomes depends on the cell origin where both cell types were cultured in StemMACs, only UCMSC secretedFGF-2 and PDGF-BB but BMMSC did not. Conclusion: Both UCMSC and BMMSC cultured in commercial serum- and xeno-free media could secret exosomes into cell culture media. These exosomes carried growth factors, including VEGF-A, FGF-2, HGF and PDGF-BB, with different expression levels.

Tumor-Derived Cell Culture Model for the Investigation of Meningioma Biology

Meningioma is the most common primary central nervous system tumor. Although mostly nonmalignant, meningioma can cause serious complications by mass effect and vasogenic edema. While surgery and radiation improve outcomes, not all cases can be treated due to eloquent location. Presently no medical treatment is available to slow meningioma growth owing to incomplete understanding of the underlying pathology, which in turn is due to the lack of high-fidelity tissue culture and animal models. We propose a simple and rapid method for the establishment of meningioma tumor-derived primary cultures. These cells can be maintained in culture for a limited time in serum-free media as spheres and form adherent cultures in the presence of 4% fetal calf serum. Many of the tissue samples show expression of the lineage marker PDG2S, which is typically retained in matched cultured cells, suggesting the presence of cells of arachnoid origin. Furthermore, nonarachnoid cells including vascular endothelial cells are also present in the cultures in addition to arachnoid cells, potentially providing a more accurate tumor cell microenvironment, and thus making the model more relevant for meningioma research and high-throughput drug screening.

Development of Poly-NIsopropylacrylamide Surfaces for the Selection of Swine Sperm

The reproductive efficiency of pig farms is directly correlated with the fertility of the boars. The aim of this work was to develop polymeric materials that can be used as a platform to select a subpopulation of sperm with better cell physiological parameters. Polymeric hydrogels composed of Poly-N-isopropylacrylamide with different positive charges given by copolymerization with (3-acrylamidopropyl) trimethylammonium chloride (APTA, 5-10-15%), were synthesized. Subsequently, the interaction between the sperm cells and the polymeric surfaces was analyzed in TALP medium. Release of the spermatozoa from the polymeric surfaces was induced by changing to Ca2+ free media. Sperm motility, cell viability, plasma membrane and acrosome integrity were evaluated. The results indicated that a higher percentage of swine sperm attached to PNIPAM co-15% APTA hydrogels (62.86±3.33%). Ninety seven percent (97.19±1.45 %) of the sperm released from the PNIPAM co-15%APTA surfaces were viable (p<0.05 vs unbound population and raw semen), with acceptable motility (58.89±1.28%) and with intact plasma and acrosomal membranes (69±1.2% and 98.5±0.65% respectively). These results indicate that hydrogels can be used to select boar sperm with high viability and mobility for use in assisted reproductive techniques.

In Vitro Culture of Bovine Fibroblasts using Select Serum-Free Media Supplemented with Chlorella vulgaris Extract

Standard cell culture practices require addition of animal-derived serum to culture media to achieve adequate cell growth. Typically, 5-10% by volume of fetal bovine serum (FBS) is used, which accounts for a vast majority of the cost of media while also imposing environmental and ethical concerns associated with the use of animal serum. Here we tested the efficacy of culturing cells by replacing serum in the media with algae extract and select additives. Using LC-MS, we compared molecular signatures of FBS to Chlorella algae extracts and identified NAD(H)/NADP(H) as common and relatively abundant features in their characteristic profiles. Bovine fibroblasts, cultured in serum-free media supplemented with C. vulgaris extract and just two growth factors plus insulin, showed significant growth with enhanced viability compared to control cells cultured without serum, albeit still lower than that of controls cultured with 10% FBS. Moreover, C. vulgaris extract enhanced cell viability beyond that of cells cultured with the two growth factors and insulin alone. These results suggest that key components in serum which are essential for cell growth may also be present in C. vulgaris extract, demonstrating that it may be used at least as a partial alternative to serum for cell culture applications.