Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson: 1614 - 1674
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| Format: | Buch |
| Sprache: | German |
| Veröffentlicht: |
Reykjavík
Verl. der Hallgrímskrirkja
1974
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XXXII, 231 S. Ill. |
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MARC
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| 245 | 1 | 0 | |a Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson |b 1614 - 1674 |c unter Beibehaltung der Dichtungsform des Orig. in dt. Sprache wiedergegeben von Wilhelm Klose |
| 264 | 1 | |a Reykjavík |b Verl. der Hallgrímskrirkja |c 1974 | |
| 300 | |a XXXII, 231 S. |b Ill. | ||
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Datensatz im Suchindex
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| adam_text | DISSERTATION
SUBMITTED TO THE
COMBINED FACULTIES FOR THE NATURAL SCIENCES AND FOR MATHEMATICS
OF THE RUPERTO-CAROLA UNIVERSITY OF HEIDELBERG, GERMANY
FOR THE DEGREE OF
DOCTOR OF NATURAL SCIENCES
PRESENTED BY
DIPLOM-BIOLOGIN ELISA KATHARINA SPECKER
BORN IN: LEIMEN
ORAL-EXAMINATION: JULY 27TH 2015
ELEKTRONISCHE VERSION UNTER _ ~
WWW.UB.UNI-HEIDEIBERE.DE/ARCHIV/ LI/
HTTP://D-NB.INFO/1075168457
IDENTIFICATION AND CHARACTERIZATION
OF POTENTIAL HUMAN EPIDERMAL STEM CELLS
REFEREES: PROF. DR. ANDREAS TRUMPP
PROF. DR. PETRA BOUKAMP
TABLE OF CONTENTS
ZUSAMMENFASSUNG VLTL
SUMMARY IX
ABBREVIATIONS
X
1 INTRODUCTION 1
1.1 THE HUMAN SKIN 1
1.1.1 DERMIS 1
1.1.2 EPIDERMIS
2
1.2 EPIDERMAL STEM CELLS 4
1.2.1 HAIR FOLLICLE
AND SEBACEOUS GLAND 4
1.2.2 THE INTERFOLLICULAR
EPIDERMIS 6
1.3 HOMEOSTASIS OF THE SKIN 9
1.4 STEM CELL NICHE 11
1.5 STUDYING EPIDERMAL STEM CELLS 12
1.6 AIM 14
2 MATERIAL AND METHODS 15
2.1 MATERIAL 15
2.1.1 CELLS 15
2.1.2 CELL CULTURE SUPPLEMENTS
AND MEDIA
15
2.1.3 ANTIBODIES, KITS
AND PRIMER 16
2.1.4 TECHNICAL EQUIPMENT
AND CONSUMABLES 18
2.1.5 BUFFER, SOLUTIONS,
CHEMICALS
AND ENZYMES 19
2.1.6 SOFTWARE 21
2.2 METHODS 22
2.2.1 CELLCULTURE
METHODS 22
2.2.1.1 CULTIVATION AND
MAINTENANCE OF HUMAN FIBROBLASTS 22
2.2.1.2 FEEDER CELLS FOR
THE COCULTIVATION WITH
KERATINOCYTES 22
2.2.1.3 ISOLATION, CULTIVATION
AND MAINTENANCE OF HUMAN
KERATINOCYTES 22
2.2.1.4 STIMULATION OF CELLS
WITH TGF|3-1 23
2.2.1.5 TREATMENT OF CELLS
WITH Y-27632 23
2.2.1.6 PROLIFERATION ASSAY 23
2.2.1.7 FREEZING AND THAWING OF
CELLS 24
2.2.1.8 MYCOPLASMTEST 24
2.2.1.9 CELL PELLETS 25
2.2.1.10 SCAFFOLD BASED
LONG TERM ORGANOTYPIC COCULTURES
(SEA
OTCS) 25
2.2.1.11 FIBROBLAST-DERIVED MATRIX
ORGANOTYPIC COCULTURES (FDM
OTCS) 26
2.2.1.12 SINGLE CELL
SUSPENSIONS OF OTCS 26
V
2.2.1.13 FACS ANALYSIS OR
SORTING 26
2.2.1.14 ISOLATION OF LRCS 27
2.2.2 PROCESSING OF TISSUE 27
2.2.2.1 PRESERVATION OF OTCS 27
2.2.2.2 CELLS ON CYTOSPINS 28
2.2.3 TET-OFF-H2B-GFP REPORTER 28
2.2.3.1 BACTERIAL TRANSFORMATION
AND PLASMID PURIFICATION 28
2.2.3.2 VIRUS PRODUCTION 28
2.2.3.3 CONCENTRATION BY ULTRACENTRIFUGATION 30
2.2.3.4 VIRUS TITRATION 30
2.2.3.5 TRANSDUCTION OF CELLS 30
2.2.4 STAINING METHODS 31
2.2.4.1 INDIRECT IMMUNOFLUORESCENCE 31
2.2.4.2 HEMATOXYLIN AND
EOSIN (H&E) - STAIN 31
2.2.5 MOLECULAR BIOLOGY
TECHNIQUES 32
2.2.5.1 RNA ISOLATION 32
2.2.5.2 DNA ISOLATION 32
2.2.5.3 REVERSE TRANSCRIPTASE 32
2.2.5.4 REAL-TIME PCR 32
2.2.6 MICROARRAY 33
2.2.7 ANALYSIS OF MICROARRAY
DATA 33
2.2.8 LINEAR AMPLIFICATION
MEDIATED (LAM)-PCR 34
RESULTS 35
3.1 OPTIMIZATION OF INFECTION METHODS FOR HUMAN ADULT PRIMARY
KERATINOCYTES WITH THE TET-OFF-H2B-GFP
LENTIVIRAL VECTOR 35
3.1.1 NHEKS CAN
BE EFFICIENTLY
INFECTED WITH
A TET-OFF-H2B-GFP LENTIVIRAL VECTOR 35
3.2 H2B-GFP EXPRESSION OF NHEKS CAN BE TIGHTLY REGULATED BY DOXYCYCLINE
37
3.3 CHARACTERIZATION OFTET-OFF-H2B-GFP POSITIVE NHEKS IN
3D OTCS 39
3.3.1 H2B-GFP TRANSDUCED NHEKS
ARE
ABLE FOR LONG-TERM
REGENERATION
IN OTCS 39
3.3.2 NO INFLUENCE
ON THE
PROLIFERATION
CAPACITY OF TRANSDUCED NHEKS IN
OTCS 40
3.3.3 REGULAR EPIDERMAL DIFFERENTIATION
OF TRANSDUCED KERATINOCYTES IN
OTCS 41
3.3.4 TIGHT
REGULATION
OF THE TET-OFF-H2B-GFP
LENTIVIRAL REPORTER WITH
DOX IN OTCS 44
3.3.5 GENE SILENCING OF H2B-GFP
POSITIVE CELLS
IN OTCS
45
3.4 LONG-TERM CULTIVATION OF NHEKS WITH
RHO KINASE INHIBITOR Y-27632 46
3.4.1 INCREASED PROLIFERATION OF NHEKS
IN THE
PRESENCE
OF Y-27632 48
3.4.2 RHO KINASE
INHIBITOR SUITABLE TO
EXPAND H2B-GFP POSITIVE NHEKS 49
3.4.3 RHO KINASE
INHIBITOR DOES
NOT SUPPORT
GROWTH
OF TRANSDUCED KERATINOCYTES IN
OTCS
50
3.5 DETECTION OF LRCS IN H2B-GFP OTCS 51
3.6 ISOLATION OF BASAL NHEKS FROM OTCS 53
3.7 ISOLATION OF VITAL LRCS 54
3.8 GLOBAL GENE EXPRESSION ANALYSIS OF ISOLATED LRCS COMPARED TO
ISOLATED BASAL CELLS OF OTCS
56
3.9 VALIDATION OF GENE ARRAY DATA BY
REAL-TIME-QUANTITATIVE-PCR 61
3.9.1 UPREGULATION OFECM GENES
IN LRCS 61
3.9.2 DEGRADATION OFECM POSSIBLY
INHIBITED IN LRCS 64
3.9.3 NOEMTINLRCS 64
3.9.4 UPREGULATION OF THE
CYTOKINE IL6 IN LRCS 64
3.9.5 DOWNREGULATION OF KERATIN
14 AND HISTONE
CLUSTERS INDICATE
REDUCED PROLIFERATION
OF LRCS 65
3.10 ROLE OF TGFB
IN LRCS 66
3.11 NO INCREASED ECM
PROTEIN EXPRESSION OF LRCS 67
3.12 INDUCTION OF A LRC PHENOTYPE
IN VITRO
BY
TGFB STIMULATION 68
3.13 TGFB PATHWAY IS ACTIVATED
UPON STIMULATION WITH
HIGH CONCENTRATIONS OF TGFB
IN NHEKS
CULTIVATED IN
LOW-CALCIUM MEDIUM 70
3.14 INFLUENCE OF TGFB ON THE
PROLIFERATION OF NHEKS IN LOW-CALCIUM MEDIUM 71
3.15 TGFB STIMULATION INCREASED ECM
PROTEIN EXPRESSION IN 2D CULTURES 73
3.16 CLONAL ANALYSIS IN OTCS 74
4 DISCUSSION 75
4.1 H2B-GFP LENTIVIRAL REPORTER FOR THE DETECTION OF LRCS 75
4.2 OTCS TO STUDY STEM CELLS IN THE HUMAN
EPIDERMIS 77
4.3 INFLUENCE OF THE RHO KINASE INHIBITOR Y-27632 ON NHEKS 78
4.4 DETECTION AND ISOLATION OF LRCS IN OTCS 79
4.5 MOLECULAR CHARACTERIZATION OF LRCS 80
4.6 A ROLE FOR TGFB SIGNALING IN LRC
REGULATION? 84
4.7 INDUCTION OF LRC-UKE PHENOTYPE BY TGFB-1 84
4.8 CLONAL KINETICS IN THE IFE 87
4.9 CONCLUSION 88
5 LITERATURE 90
6 APPENDIX 102
|
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| author | Hallgrímur Pétursson 1614-1674 |
| author2 | Klose, Wilhelm |
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| author_facet | Hallgrímur Pétursson 1614-1674 Klose, Wilhelm |
| author_role | aut |
| author_sort | Hallgrímur Pétursson 1614-1674 |
| author_variant | h p hp |
| building | Verbundindex |
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| indexdate | 2024-07-09T22:26:56Z |
| institution | BVB |
| language | German |
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| physical | XXXII, 231 S. Ill. |
| publishDate | 1974 |
| publishDateSearch | 1974 |
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| publisher | Verl. der Hallgrímskrirkja |
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| spelling | Hallgrímur Pétursson 1614-1674 Verfasser (DE-588)118761587 aut Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 unter Beibehaltung der Dichtungsform des Orig. in dt. Sprache wiedergegeben von Wilhelm Klose Reykjavík Verl. der Hallgrímskrirkja 1974 XXXII, 231 S. Ill. txt rdacontent n rdamedia nc rdacarrier (DE-588)1071854844 Fiktionale Darstellung gnd-content Klose, Wilhelm trl DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=019745976&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hallgrímur Pétursson 1614-1674 Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 |
| subject_GND | (DE-588)1071854844 |
| title | Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 |
| title_auth | Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 |
| title_exact_search | Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 |
| title_full | Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 unter Beibehaltung der Dichtungsform des Orig. in dt. Sprache wiedergegeben von Wilhelm Klose |
| title_fullStr | Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 unter Beibehaltung der Dichtungsform des Orig. in dt. Sprache wiedergegeben von Wilhelm Klose |
| title_full_unstemmed | Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson 1614 - 1674 unter Beibehaltung der Dichtungsform des Orig. in dt. Sprache wiedergegeben von Wilhelm Klose |
| title_short | Die Passionspsalmen des isländischen Dichters Hallgrímur Pétursson |
| title_sort | die passionspsalmen des islandischen dichters hallgrimur petursson 1614 1674 |
| title_sub | 1614 - 1674 |
| topic_facet | Fiktionale Darstellung |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=019745976&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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