isolation and growth of stem cells

... mutant cells and extent of HIV Growth Growth rates of parental and mutant cells and extent of HIV integration (A) Growth rates Parental and mutant 30-2 and 42-7 cells were seeded and growth measured ... x 105 Figure cells with an HIV-EGFP vector at a moi = 0.5 Infection of parental and mutant HeLa cells (30-2 and 42-7) Infection of parental and mutant HeLa cells (30-2 and 42-7) cells with an ... fractionation of cytoplasmic and nuclear extracts and measurement of the amount of full product 36 hours after infection in parental (black bars) and 42-7 (grey bars) cells; (E) Parental and 42-7 cells...

Ngày tải lên: 13/08/2014, 05:22

... glioma cells and re-expression of miR-26b diminished proliferation, migration and invasion of glioma cells in vitro (25) Moreover, miR-26b targets ephrin A2 and diminish the capacity of glioma cells ... glioma cells enhances its viability, soft-agar colony forming ability and invasiveness (33) Silencing of miR-335 causes growth arrest and apoptosis as well as reduced invasion and reduced growth of ... -3- Table of Contents Table of Contents Summary List of Tables List of Figures List of Abbreviations 10 Chapter 1: Introduction 13 1.1 Discovery, biogenesis and mechanisms of action of microRNAs...

Ngày tải lên: 09/09/2015, 17:53

... potency of stem cells 1.1.2 Fundermental features of stem cells 1.1.3 Signalling pathways modulating pluripotency in stem cells 1.1.4 Transcription factors controlling pluripotency of stem cells ... Developmental potency of stem cells Stem cells are unspecialized cells that have the capability of self-renewal for producing identical unspecialized daughter cells and the potential of differentiation ... specialized cells Stemness refers to the undifferentiated status of self-renewing stem cells Stem cells show a hierarchy of developmental potency in mouse (Fig 1-1) Totipotent stem cells include...

Ngày tải lên: 14/09/2015, 13:38

... (Figs 4E and F) Discussion Isolation and characterization of stem cells derived from various tissues and sources have been one very critical issue for stem cell therapy [12,28,33] The purpose of this ... Cotransplant of neural stem cells and NT-3 gene modified Schwann cells promote the recovery of transected spinal cord injury Spinal Cord 2007, 45, 15-24 12 Hipp J, Atala A Sources of stem cells for ... measured and calculated and drawn as a graph A Fig Identification of the cumulative population doubling level (CPDL) and culture of canine umbilical cord blood (cUCB)-mesenchymal stem cells (MSCs) Cells...

Ngày tải lên: 07/08/2014, 23:22

... during injury, stem cells can adopt this form of cell division to expand and replenish the depleted stem cell pool (Morrison and Kimble, 2006) 1.4 1.4.1 Types of stem cells Embryonic stem cells During ... 1.3 Symmetrical and asymmetrical division 1.4 Types of stem cells 1.4.1 Embryonic stem cells 1.4.2 Somatic stem cells 1.4.3 Stem cells and cancer ... maintain these cells in an undifferentiated, quiescent state For stem cells to be activated, the niche environment must change (Fuchs, 2007) 1.4.3 Stem cells and cancer Stem cells and cancer cells share...

Ngày tải lên: 12/09/2015, 21:26

... acetylation of histone H4; H3K4me1/2/3, (mono-, di- and tri) methylation of lysine of histone H3; H3K27ac, acetylation of lysine 27 of histone H3; H3K27me3, trimethylation of lysine 27 of histone ... Figure The combination of stem cell models and epigenomics in studies of the role of non-coding mutations in human disease Epigenomic analyses of cells derived through in vitro stem cell differentiation ... embryonic stem cell; H3K4me1, methylation of lysine of histone H3; H3K4me3, trimethylation of lysine of histone H3; H3K27ac, acetylation of lysine 27 of histone H3; H3K27me3, trimethylation of lysine...

Ngày tải lên: 11/08/2014, 12:21

... my two supervisors Prof Teoh Swee-Hin and A/Prof Jerry Chan I would like to thank them for their constant sharing of knowledge and ideas, their patience and understanding, and the many hours they ... the main characteristics of mature endothelial cells and its progenitor cells Table 2-10 Comparison between UCB and adult PB-EPC in terms of its colony emergence, growth and vessel forming ability ... the upregulation of several angiogenic genes and secretion of growth factors Figure reproduced from Carmeliet and Jain (Carmeliet and Jain 2000) (B) Sprouting of the endothelial cells is induced,...

Ngày tải lên: 09/09/2015, 10:16

... polysaccharide One milligram of the partially purified polymer contained 309.7 µg of protein, 160.2 µg of neutral sugar, 58.5 µg of hexosamine, 30.8 µg of uronic acid and 3.8 µg of phosphorus Constituent ... concentration of ppm to estimate the effect of pH After min, A660 was measured and the extent of flocculation was calculated Pronase digestion of extracellular polymer Forty milligrams of the extracellular ... as follows: 500 g of soil was washed times with distilled water and suspended in 1,000 ml of water The mixture was allowed to stand for 10min and the supernatant was collected and used for the...

Ngày tải lên: 05/09/2013, 09:08

... III and P IV isoforms from the eluate P III ⁄ P IV (Fig 1C) Isoelectric focusing gave pI of 5.8, 5.7, 5.5 and 5.2 for the four isoforms respectively Secondary structure of the pulchellin isoforms ... mixture of other isoforms (B) Gel visualization of proteins eluted Lane 1, molecular weight markers Numbers on the left indicate the Mr values of the standards in thousands Lanes 2–9, lg of peak ... P III ⁄ P IV and a mixture of other isoforms (*), respectively, in the presence (lanes 2–5) or absence (lanes 6–9) of 2-mercaptoethanol (C) Elution profile of P III and P IV Chromatofocusing chromatography...

Ngày tải lên: 18/02/2014, 16:20

... of 50 nmÆmin)1, band-width of 1.0 nm, and a resolution of 0.1 nm in the temperature range between 10 °C and 70 °C, after preincubation for 10 at each temperature Three runs were accumulated and ... of 0.6 lM The data were fitted to Michaelis–Menten curves Squares and dotted line: data and fitting with PHI/PHO ratio of 0.5 Triangles and continuous line: data and fitting with PHI/PHO ratio of ... [19] Cells were harvested when growth reached the stationary phase and were stored frozen ()80 °C) Preparation of crude extract Cells were washed twice in 50 mM Hepes/NaOH buffer, pH 7.0, and...

Ngày tải lên: 21/02/2014, 00:20

... intense band of 61 kDa (NrfA) and a band of weak intensity of 19 kDa (NrfH), confirming its hetero-oligomeric nature (Fig 1, lane 1) However, in the absence of boiling (Fig 1A, lanes and 4) high ... and 4) high molecular mass bands of approximately 110 kDa and > 200 kDa were visible, as well as a faint band at 37 kDa, suggesting the presence of dimers All of the bands stained positively for ... between NrfH and NrfA is discussed The reassessment of previous spectroscopic studies was undertaken, particularly regarding the assignment of spectroscopic and redox potentials of the NrfA hemes...

Ngày tải lên: 21/02/2014, 00:20

... total of mg of curdione (1), mg of curcumol (2), 10 mg of germacrone (3), 18 mg of curzerene (4), mg of 1,8-cineole (5) and 17 mg of b-elemene (6) from the essential oil (300 mg) in 2.4 mL of the ... the differences between K2 and K3, and between K4 and K5 in solvent systems (petroleum ether/acetonitrile/acetone 2:1:1 and 4:3:1 v/v/v) are 0.34 and 0.24, and 0.20 and 0.20, respectively, which ... Both of the upper and the lower phases were degassed for 30 by ultrasonication before use Aliquots of 300 mg of the essential oil were diluted by 2.4 mL of the mixed solution of lower phase and...

Ngày tải lên: 07/03/2014, 21:20

... Chlamydomonas cells Chlamydomonas CW15 cells were grown to a cell density > ´ 106 cells per mL Cells were pelleted, resuspended in 30 mM Tris/HCl pH 7.9, and broken by two cycles of freeze-thawing ... BLAST of peptide sequences of 21 kDa proteins in databases Tryptic peptides were puri®ed by RP-HPLC and some of them were totally or partially analyzed by Edman sequencing (ááá) and/ or by MALDI-TOF ... BAS1 protein of Brassica, spinach, barley and A thaliana, PR1 of Phaseolus and MHF of A thaliana These proteins belong to the 2Cys-Prx subfamily All plant 2Cys-Prx proteins, except BAS1 of barley,...

Ngày tải lên: 08/03/2014, 16:20

... presequence of 19 amino acids The mature protein would have a molecular mass of 41.7 kDa and a pI of 6.34 The YedO protein from E coli and the d-CDes from Arabidopsis showed an overall identity of 36% and ... nutritional value of plant species short of S-containing amino acids By producing transgenic d-CDes plants this goal might be reached Experimental procedures Growth and harvest of plants Seeds of Arabidopsis ... purity of mitochondria prepared from green tissue of tobacco, pea and Arabidopsis thaliana Phyton 43, 215–229 53 Leegood RC & Malkin R (1986) Isolation of sub-cellular photosynthetic systems In...

Ngày tải lên: 16/03/2014, 18:20

... respectively, and the resultant VNARs evaluated for retention of binding affinity Material and methods Equipment and reagents Restriction enzymes and Vent DNA polymerase were purchased from New England ... ampicillin and/ or 2% (w/v) glucose Solid media contained 2% (w/v) Bacto-agar Transformation of E coli was by standard procedures [24] performed using electrocompetent cells Isolation of total RNA ... possibility of inter- and intra CDR disulphide cross-links These strategies are summarized in Fig 1B and Table 1, and details of the library construction are given in greater detail in the Materials and...

Ngày tải lên: 17/03/2014, 10:20

... thickness of *6.5 nm Combining the structural analysis of TEM and AFM, the as-prepared hematite particles are of nanodiscs in 123 J Nanopart Res Particle nucleation and growth Fig a TEM image of hematite ... absence of surfactants under mild conditions, and no need of toxic precursors and high-temperature decomposition in organic solvents The nanodiscs (diameters of *50 nm and thickness of *6.5 nm) ... addition of phosphate ions during the hydrolysis and growth processes Reeves and Mann (1991) suggested that the interaction of phosphate with hematite crystals was not specific to a single set of symmetryrelated...

Ngày tải lên: 20/03/2014, 13:08

... re-testing and validating the effects on cell growth by transforming W303.1C and plating the cells at 23 °C, 30 °C and 37 °C A cold-sensitive growth phenotype (where toxin is active and interferes ... cold-sensitive growth phenotype seen in this yeast strain genuinely reflects of the sensitivity of the mutant toxin to temperature To obtain a clearer picture of the growth profiles of yeast cells expressing ... exception of 37 °C In contrast, the Kar2SP-RTAD variant showed comparable growth at all temperatures The growth profiles of Kar2SPRTAE177D at 30 °C and 37 °C, and Kar2SPRTAF108L ⁄ L151P at 23 °C and...

Ngày tải lên: 23/03/2014, 07:20

... Switzerland), thermolysin, chymotrypsin or elastase (Sigma, St Louis, MO, USA)] and Cloning of the bovine MUC15 cDNA by PCR with degenerate primers Isolation of total RNA from the mammary gland of ... PCR screening of bovine and human cDNA libraries showed the presence of MUC15 mRNA in human breast, bovine mammary gland, bovine lymph nodes and lungs of both species (Table 1) Of the identified ... examined and specific PCR primers were designed enclosing the coding sequence of the bovine protein To investigate the presence of MUC15 expression in epithelial cells of the human mammary gland,...

Ngày tải lên: 24/03/2014, 04:21

... of degenerate and specific primers and by rescreening the cDNA libraries The partial clones of XHIVEP2 and -3 covered 5.9 and 3.2 kb of the respective 3¢ ends and included 3¢-UTRs and poly(A)-tails ... upper strands with [32P]ATP[cP] and T4 polynucleotide kinase The labeled oligomers were annealed by heating to 90 °C an equimolar mixture of the upper and lower strands in reaction buffer and cooling ... -2 and -3) The Xenopus XHIVEPs are characterized with respect to temporal and spatial expression, nuclear import/export activity and DNA binding specificity Materials and methods Isolation and...

Ngày tải lên: 30/03/2014, 13:20

... B-3 and CCL-75 cells ranged from 40% to 70% To determine the effect of wildtype and mutant forms of GST on injected cells, all surviving HLE B-3 and CCL-75 cells were scored at 24 and 48 h and ... designated C4, D7 and The anchorage-independent growth of phenotypically transformed cells was confirmed by assay of soft agar colonies [26] Clone C4 cells grew into colonies within weeks of plating, ... characterization of hGSA4-transfected cells (A) Phenotypic transformation of hGSA4-transfected cells (a) Control WT-HLE B-3 cells; (b) HLE B-3 cells weeks after hGSTA4 transfection; (c) growth arrest and...

Ngày tải lên: 30/03/2014, 13:20