Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells:
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1. Verfasser: | |
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Format: | Abschlussarbeit Buch |
Sprache: | English |
Veröffentlicht: |
2015
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Schlagworte: | |
Online-Zugang: | Inhaltsverzeichnis |
Beschreibung: | XII, 108 S. Ill., graph. Darst. |
Internformat
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Datensatz im Suchindex
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adam_text | TABLE OF CONTENTS
LIST OF FIGURES VIII
LIST OF TABLES X
LIST OF ABBREVIATIONS XII
1. INTRODUCTION 1
1.1 PRINCIPLES UNDERLYING MAMMALIAN NEURAL INDUCTION 1
1.2. NEUROEPITHELIAL CELLS IN THE DEVELOPING CENTRAL NERVOUS SYSTEM 2
1.2.1 CELLULAR JUNCTIONS OF NEUROEPITHELIAL CELLS 3
1.2.2 THE EPITHELIAL NATURE OF NEUROEPITHELIAL CELLS 4
1.3 ESTABLISHMENT OF CELL POLARITY IN MAMMALIAN CELLS 5
1.3.1 DISRUPTION OF CELLULAR SYMMETRY 5
1.3.2 ASYMMETRIC ASSEMBLY OF CELL-CELL CONTACTS AND POLARITY PROTEINS 6
1.3.3 REARRANGEMENT OF THE CYTOSKELETON 6
1.3.4 APICAL VESICULAR TRAFFICKING AND MAINTENANCE OF APICO-BASAL
POLARITY 7
1.4 PRINCIPLES OF TUBE FORMATION 8
1.5 NEURAL TUBE FORMATION 9
1.5.1 CELLULAR MECHANISMS UNDERLYING PRIMARY NEURULATION 10
1.5.2 MOLECULAR MECHANISMS UNDERLYING PRIMARY NEURULATION 11
1.5.3 CELLULAR MOVEMENTS UNDERLYING SECONDARY NEURULATION 12
1.6 PLURIPOTENT STEM CELLS 13
1.7 NEURAL DIFFERENTIATION OF HUMAN EMBRYONIC STEM CELLS 13
1.7.1 NEURAL ROSETTES 15
1.8 ROSETTE FORMATION DURING ORGANOGENESIS AND DEVELOPMENT 16
1.9 LYSOPHOSPHATIDIC ACID 18
1.9.1 LPA SIGNALING IN NPC S OF THE CENTRAL NERVOUS SYSTEM 18
1.9.2 LPA INDUCES ROSETTE FORMATION IN CULTURED AXOLOTL NPC S 19
1.10 AIMS OF THE STUDY 21
HTTP://D-NB.INFO/1073211576
2. RESULTS - CHAPTER I
ESTABLISHMENT OF A HUMAN ES CELL-BASED NEURAL DIFFERENTIATION
PROTOCOL TO INVESTIGATE THE ROSETTE FORMATION POTENTIAL OF NEURAL
PROGENITOR CELLS
23
2.1 DIFFERENTIATION OF HUMAN EMBRYONIC STEM CELLS INTO NEURAL PROGENITOR
CELLS 23
2.1.1 HUMAN EMBRYONIC STEM CELL CULTURE 24
2.2 DEVELOPMENT OF A THREE-DIMENSIONAL NEUROEPITHELIAL CYST-BASED NEURAL
DIFFERENTIATION PROTOCOL 25
2.2.1 PAX6
+
NEURAL PROGENITOR CELLS CAN BE PASSAGED AND REORGANIZE IN AN EPITHELIAL-
LIKE LAYER OF CELLS 27
2.2.2 SERUM AND LPA TREATMENT DO NOT INDUCE LARGE ROSETTE FORMATION IN
PAX6
+
HUMAN NEURAL PROGENITOR CELLS 28
2.2.3 DERIVATION OF SOXL
+
- AND ROSETTE-FORMING NEURAL PROGENITOR CELLS 29
2.2.4 SERUM AND LPA INDUCE LARGE ROSETTE FORMATION IN SOXL
+
NEURAL PROGENITOR CELLS
31
2.3 DEVELOPMENT OF A 3D DUAL SMAD SIGNALING INHIBITION PROTOCOL 32
2.3.1 HESC-DERIVED ROSETTE NEURAL PROGENITOR CELLS EXPRESS SOXL AND
MAINTAIN THEIR
ROSETTE IDENTITY AFTER PASSAGING 33
2.3.2 LPA INDUCES LARGE ROSETTE FORMATION IN SOXL
+
NPC S AT EARLY PASSAGING
NUMBERS 34
2.4 CHARACTERIZATION OF LPA-INDUCED ROSETTE-LIKE STRUCTURES 36
2.4.1 ROSETTE-FORMING NEURAL PROGENITOR CELLS EXHIBIT APICAL TO BASAL
POLARITY AND
EXPRESS EARLY NEUROEPITHELIAL MARKERS 36
2.4.2 LPA INCREASES ROSETTE LUMEN SIZES IN A CONCENTRATION DEPENDENT
MANNER 38
2.4.3 APPLICATION OF THE CELLPROFILER SOFTWARE AS A TOOL TO ANALYZE
DIFFERENT ROSETTE
PARAMETERS 39
2.4.4 CELL PROFILER BASED PARAMETRICAL IMAGE ANALYSIS OF DIFFERENT
ROSETTE SIZES 41
2.4.5 LPA DOES NOT INCREASE CELL PROLIFERATION IN HUMAN NPC S 43
2.4.6 CELL PROLIFERATION IS NOT REQUIRED FOR LARGE ROSETTE FORMATION 44
2. RESULTS - CHAPTER II
INVESTIGATION OF THE CELLULAR MECHANISMS UNDERLYING LPA-INDUCED
LUMEN FORMATION AND EPITHELIALIZATION 47
2.5 LPA DYNAMICALLY REGULATES THE EPITHELIAL ORGANIZATION OF NEURAL
PROGENITOR CELLS . 47
2.6 ELECTRON MICROSCOPY REVEALS CHANGES IN THE EPITHELIAL ORGANIZATION
OF NEURAL
PROGENITOR CELLS DURING LPA EXPOSURE 49
2.7 LPA IS ELIMINATED FROM THE CULTURE MEDIUM WITHIN 10 HOURS 50
2.8 LPA INCREASES THE AMOUNT OF EPITHELIALIZED AREA IN A
CONCENTRATION-DEPENDENT
MANNER 52
2. 9 LPA INCREASES APICAL DOMAIN SIZE OF NPC S 54
2.10 LARGE ROSETTE-FORMING NEURAL PROGENITOR CELLS EXHIBIT DIFFERENT
APICAL DOMAIN
SIZES 56
2.11 A TIME COURSE REVEALS CHANGES IN THE EPITHELIAL MORPHOLOGY OF NPC S
IN RESPONSE
TO DIFFERENT LPA CONCENTRATIONS 58
2. RESULTS - CHAPTER III
MOLECULAR MECHANISMS UNDERLYING LPA-INDUCED LUMEN
FORMATION AND EPITHELIALIZATION 60
2.12 LPA RECEPTOR EXPRESSION IN HUMAN-DERIVED NEURAL PROGENITOR CELLS 60
2.13 RHO-SIGNALING IS REQUIRED FOR EPITHELIALIZATION AND LARGE ROSETTE
FORMATION 61
2.14 BLOCKAGE OF LPA RECEPTOR SIGNALING DECREASES LARGE ROSETTE SIZE 62
3. DISCUSSION 65
3.1 DIFFERENT NEURAL PROGENITOR POPULATIONS EXPRESS DIFFERENT NEURAL
TRANSCRIPTION
FACTORS AND EXHIBIT DIFFERENT MORPHOLOGIES 65
3.2 SERUM AND LPA INDUCE EPITHELIAL CHANGES AND LARGE ROSETTE FORMATION
IN SOXL
+
NEURAL PROGENITOR CELLS 66
3.3 LPA AND ITS INFLUENCE ON THE EPITHELIAL MORPHOLOGY OF NEURAL
PROGENITOR CELLS 68
3.4 LARGE ROSETTE FORMATION AS A CELLULAR TOOL, TO INVESTIGATE
PARAMETERS UNDERLYING
LUMEN FORMATION 71
3.5 POTENTIAL BIOLOGICAL ROLES FOR LPA 72
3.6 SUMMARY 76
4. MATERIALS AND METHODS 78
4.1 HUMAN ES CELL- AND NEURAL PROGENITOR CELL MEDIA 80
4.2 HUMAN ES CELL CULTURE 81
4.2.1 DETAILED PROTOCOL FUR HUMAN ES CELL PASSAGING AND MAINTENANCE 82
4.3 NEURAL DIFFERENTIATION 83
4.3.1 DIFFERENTIATION OF HUMAN ES CELLS INTO NEUROEPITHELIAL CYSTS 84
4.3.2 DETAILED PROTOCOL FOR NEUROEPITHELIAL CYST FORMATION AND CYST
PLATING 84
4.3.3 PROTOCOL OF NEUROEPITHELIAL CYST-DERIVED ROSETTE FORMATION 85
4.3.4 DETAILED PROTOCOL FOR NEUROEPITHELIAL ROSETTE FORMATION 85
4.3.5 DUAL SMAD INHIBITION NEURAL DIFFERENTIATION PROTOCOL 86
4.3.6 DETAILED PROTOCOL FOR DUAL SMAD SIGNALING INHIBITION 86
4.4 PROTOCOL FOR THE PASSAGING OF NEURAL PROGENITOR CELLS 88
4.4.1 DETAILED PROTOCOL FOR THE PASSAGING OF NEURAL PROGENITOR CELLS 88
4.4.2 LPA EXPOSURE TO NEURAL PROGENITOR CELLS AND LARGE ROSETTE
FORMATION ASSAY... 89
4.5 IMMUNOCYTOCHEMISTRY 90
4.5.1 PRIMARY- AND SECONDARY ANTIBODIES 91
4.6 MICROSCOPY 92
4.7 ELECTRON MICROSCOPY 92
4.8 RT-PCR 92
4.9 QUANTITATIVE PCR 93
4.10 VIRAL PRODUCTION 94
4.11 VIRAL TRANSFECTION OF HUMAN ES CELLS 95
4.12 STATISTICAL ANALYSIS 95
ACKNOWLEDGEMENTS 96
REFERENCES 97
|
any_adam_object | 1 |
author | Medelnik, Jan-Philip 1983- |
author_GND | (DE-588)1073227804 |
author_facet | Medelnik, Jan-Philip 1983- |
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dewey-full | 573.8638619353 |
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dewey-ones | 573 - Specific physiological systems in animals |
dewey-raw | 573.8638619353 |
dewey-search | 573.8638619353 |
dewey-sort | 3573.8638619353 |
dewey-tens | 570 - Biology |
discipline | Biologie |
format | Thesis Book |
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spelling | Medelnik, Jan-Philip 1983- Verfasser (DE-588)1073227804 aut Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells von Medelnik, Jan-Philip 2015 XII, 108 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Dresden, Techn. Univ., Diss., 2015 Vorläuferzelle (DE-588)4418410-4 gnd rswk-swf Lysophosphatidsäuren (DE-588)4381995-3 gnd rswk-swf Nervenzelle (DE-588)4041649-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Nervenzelle (DE-588)4041649-5 s Vorläuferzelle (DE-588)4418410-4 s Lysophosphatidsäuren (DE-588)4381995-3 s DE-188 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028096939&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
spellingShingle | Medelnik, Jan-Philip 1983- Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells Vorläuferzelle (DE-588)4418410-4 gnd Lysophosphatidsäuren (DE-588)4381995-3 gnd Nervenzelle (DE-588)4041649-5 gnd |
subject_GND | (DE-588)4418410-4 (DE-588)4381995-3 (DE-588)4041649-5 (DE-588)4113937-9 |
title | Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells |
title_auth | Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells |
title_exact_search | Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells |
title_full | Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells von Medelnik, Jan-Philip |
title_fullStr | Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells von Medelnik, Jan-Philip |
title_full_unstemmed | Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells von Medelnik, Jan-Philip |
title_short | Cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells |
title_sort | cellular and molecular effects of lysophosphatidic acid on cultured human neural progenitor cells |
topic | Vorläuferzelle (DE-588)4418410-4 gnd Lysophosphatidsäuren (DE-588)4381995-3 gnd Nervenzelle (DE-588)4041649-5 gnd |
topic_facet | Vorläuferzelle Lysophosphatidsäuren Nervenzelle Hochschulschrift |
url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028096939&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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