LIFE09200604007 Tabish - Homi Bhabha National Institute

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Results Fig. 10: Cell population doubling time for 6 representative cell lines at 6 time points (0-96 h). The doubling time ranged from 12 h to 36 h with an average of approximately 24 h. Each point represents mean value of four different readings. Bars indicate standard error of mean. The average PD of few representative LCLs was found to be 24 h, ranging from 12 h to 36 h (Fig. 10). The LCLs grew as clusters exhibiting typical rosette morphology in suspension cultures, but single cells were also observed having big nucleus and numerous vacuoles (Fig.6). EBV is known to specifically infect B cells allowing their growth in culture hence the transformed phenotype is expected to have homogeneous cell population, however flow cytometric analysis revealed the presence of dual population (Fig. 11). On the basis of morphology and granularity it was revealed that lower (R1) population represents mono cell suspension of interest and the other (R2) population probably represents fraction of cells that have spontaneously differentiated into smaller lymphoid cells with shrunken nucleus that ultimately undergo apoptosis during conventional cell culture thus representing diverse size and granularity. Out of 34 cell lines 4 MPN patient cell lines did not grow efficiently in the continuous culture, hence were not undertaken for any phenotypic assays. 90
Results Fig. 11: Morphological analysis of established lymphoblastoid cell lines. Light microscopy image of LCL showing typical rosette morphology. Cells grow in clumps while single cells are also seen (a). Confocal microscopy image of cells in EBV transformed cell lines showing big nucleus and numerous vacuoles (b). Flow cytometry cluster plot showing two distinct populations in LCL where lower R1 population represents mono cell suspension and R2 population represents cell aggregates (c). 4.1.3 Cell surface marker EBV specifically infects B cells hence immunophenotyping for B cell specific marker was done to ensure pure population. Antibodies against B (CD19), T (CD3) and NK (CD56) cell surface markers were used. Data confirmed that cells from representative randomly selected LCLs showed expression of typical B cell surface marker (CD19) while markers for T cell (CD3) and NK cells (CD56) were absent (Fig. 12), thus ascertaining the purity of growing cultures. 4.1.4 DNA ploidy analysis DNA ploidy status of the LCLs was assessed immediately after cell line preparation at low population doubling (
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Genotype-Molecular Phenotype Correl
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CONTENTS Page No Synopsis 1 List of
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Synopsis
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Synopsis Genotype-Molecular Phenoty
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Synopsis Materials and Methods: 1.
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Synopsis loading control. PCR produ
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Synopsis 7.1 Percent cell death aft
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Synopsis slides were then dried and
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Synopsis range 41.77% - 90.95% at 5
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Synopsis 5. Genotyping of genes: Ge
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Synopsis G2/M arrest and delaying e
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Synopsis may have an important bear
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Synopsis References: 1. Bobba, R.;
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Abbreviations MOPS : 3-(N-morpholin
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List of Figures Fig. 28: Percent re
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Chapter 1 Introduction
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Introduction In previous studies fr
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Introduction important carcinogenes
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Review of Literature The yet undeci
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Review of Literature Fig. 2: Incide
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Review of Literature sustained angi
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Review of Literature 2.6 Genotype p
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Review of Literature 2.7.2 Biologic
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Review of Literature Table 1: Steps
Page 53 and 54: Review of Literature future analysi
Page 55 and 56: Review of Literature Inefficient ce
Page 57 and 58: Review of Literature XRCC1 (X-ray r
Page 59 and 60: Review of Literature 2.9.2 Gene inv
Page 61 and 62: Review of Literature MPO gene polym
Page 63 and 64: Review of Literature important role
Page 65 and 66: Materials and Methods Source of che
Page 67 and 68: Materials and Methods Method: EBV-t
Page 69 and 70: Materials and Methods Immunophenoty
Page 71 and 72: Materials and Methods with 500 μl
Page 73 and 74: Materials and Methods cDNA it can b
Page 75 and 76: Materials and Methods Cobalt-60 iso
Page 77 and 78: Materials and Methods specific bind
Page 79 and 80: Materials and Methods h half of the
Page 81 and 82: Materials and Methods phenol and th
Page 83 and 84: Materials and Methods 7. Filter ste
Page 85 and 86: Materials and Methods viz. EXO-SAP
Page 88 and 89: Materials and Methods Table 2: List
Page 90 and 91: Materials and Methods SMAD6 AREG HM
Page 92 and 93: Materials and Methods 3.9 Effect of
Page 94 and 95: Materials and Methods fresh eppendo
Page 96 and 97: Materials and Methods Power SYBR Gr
Page 98 and 99: Results Objective 1 To generate EBV
Page 100 and 101: Results Fig. 8: Frequency of second
Page 102 and 103: Results Table 6: Demographics of he
Page 106 and 107: Results Fig. 12: Cell surface marke
Page 108 and 109: Results 4.1.5 Expression of ATM gen
Page 110 and 111: Results Objective 2 To compare the
Page 112 and 113: Results Fig. 17: Standardization of
Page 114 and 115: Results Fig. 19: Standardisation of
Page 116 and 117: Results The cells were incubated fu
Page 118 and 119: Results 4.2.3.1 Percent G2 delay af
Page 120 and 121: Results 4.2.3.2 Effect of BPDE expo
Page 122 and 123: Results Fig. 29: Comparison of perc
Page 124 and 125: Results Fig. 31: Percent cell death
Page 126 and 127: Results normalization with baseline
Page 128 and 129: Results 4.2.5.2 Real time PCR valid
Page 130 and 131: Results homozygous wild-type, heter
Page 132 and 133: Results Fig. 36b: Electrophoresis o
Page 134 and 135: Results Fig. 37a: Representative el
Page 136 and 137: Results Table 13: Consolidated geno
Page 138 and 139: Results LCL Genes NAT2(1) NAT2(2) N
Page 140 and 141: Results Fig. 38: Distribution of Ge
Page 142 and 143: Results Fig. 39: Various combinatio
Page 144 and 145: Results Table 14: Genotype-phenotyp
Page 146 and 147: Discussion Introduction Squamous ce
Page 148 and 149: Discussion stimulated lymphocytes w
Page 150 and 151: Discussion H2AX is currently the mo
Page 152 and 153: Discussion or G2/M phase arrest. As
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Discussion differential expression
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Discussion DNA repair genes and MPN
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Chapter 6 Summary and Conclusions
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Summary and Conclusions Statistical
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Bibliography 1. Dikshit, R.; Gupta,
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Bibliography 43. Lei, D.; Sturgis,
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Bibliography 76. Fry, R. C.; Svenss
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Bibliography 115. Kote-Jarai, Z.; M
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Bibliography 149. Hashibe, M.; Bren
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Bibliography 183. Bergamaschi, D.;
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Bibliography 216. Bondy, M. L.; Wan
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Indian J Med Res 135, June 2012, pp
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822 INDIAN J MED RES, JUNE 2012 mar
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824 INDIAN J MED RES, JUNE 2012 (h)
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826 INDIAN J MED RES, JUNE 2012 Tab
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828 INDIAN J MED RES, JUNE 2012 lik
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CASE REPORT Eben L. Rosenthal, MD,
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FIGURE 2. Chromosomal breakage anal
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FA along with ataxia telangiectasia
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LIFE09200604007 Tabish - Homi Bhabha National Institute

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