Vascular growth: a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2012
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | Getr. Zählung Ill., graph. Darst. |
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Record in the Search Index
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| adam_text | IMAGE 1
TABLE OF CONTENTS
I
I. 7A6CE O F CONTENTS
1. T A B L E O F C O N T E N T S 1
I I . T A B L E O F F I G U R E S V
I I I . T A B L E O F T A B L E S V I
I V . T A B L E O F A B B R E V I A T I O N S V I I
L I T E R A T U R E L-L
A P P E N D I X A - X V I I I
1 . I N T R O D U C T I O N 1
1 . 1 . THE CARDIOVASCULAR SYSTEM 2
1 . 1 . 1 . ATHEROSCLEROSIS - CAUSE FOR CLINICAL CARDIOVASCULAR
COMPLICATIONS 3
1 . 1 . 2 . A U T O N O M O U S COMPENSATORY VASCULAR GROWTH PROCESSES 6
1 . 2 . THERAPIES FOR ISCHEMIA-RELATED DISEASES 9
1 . 2 . 1 . CARDIOVASCULAR COMPLICATIONS AND THEIR CURRENT TREATMENT 9
1 . 2 . 2 . CARDIOVASCULAR COMPLICATIONS AND CYTOKINE-BASED THERAPIES 1
1
1 . 2 . 2 . 1 . PROMINENT MOLECULES F O R CYTOKINE-BASED THERAPIES 1 3
1 . 2 . 2 . 2 . THE CYTOKINE G-CSF 1 4
1 . 2 . 3 . CARDIOVASCULAR COMPLICATIONS AND CELL-BASED THERAPIES 1 6
1 . 2 . 3 . 1 . M A J O R CELLS F O R CELL-BASED THERAPIES - ENDOTHELIAL
CELLS AND THEIR PROGENITORS 1 8
1 . 2 . 3 . 1 . 1 . CHARACTERISATION O F ENDOTHELIAL CELLS AND THEIR
PROGENITORS 1 8
1 . 2 . 3 . 1 . 2 . SOURCES F O R ENDOTHELIAL PROGENITOR CELLS 1 9
1 . 2 . 3 . 1 . 3 . ENDOTHELIAL PROGENITOR CELLS AND THEIR APPLICATION 2
0
2 . P U R P O S E O F T H E S T U D Y 2 2
HTTP://D-NB.INFO/1031456279
IMAGE 2
TABLE OF CONTENTS
II
3 . M A T E R I A L S & M E T H O D S 2 5
3 . 1 . MATERIALS 2 5
3 . 1 . 1 . CHEMICALS 2 5
3.1.2. MATERIALS AND EQUIPMENT 28
3 . 1 . 3 . BUFFERS 3 2
3 . 1 . 4 . ENZYMES 3 3
3.1.5. ANTIBODIES 33
3 . 1 . 6 . CHEMOKINES 3 4
3 . 1 . 7 . READY TO USE KITS 3 5
3.1.8. PCR-MARKER 35
3.1.9. PCR-PRIMERS 36
3 . 1 . 1 0 . M E D I A 3 6
3 . 1 . 1 1 . CELL LINES 3 9
3.1.11.1. BEND3 - ENDOTHELIAL CELLS 39
3.1.11.2. C6 - GLIOMA CELLS 39
3.1.11.3. GPSCS - PLURIPOTENT CELLS 39
3.1.11.4. HUVECS - HUMAN ENDOTHELIAL CELLS 39
3.1.11.5. H T B 5 6 - T U M O U R CELLS 39
3.1.11.6. O P 9 - S T R O M A L CELLS 40
3.1.12. ANIMALS 4 0
3.1.12.1. C57BL6 MICE 40
3.1.12.2. NMRI/NUDE MICE 40
3.1.12.3. ANIMAL HUSBANDRY 40
3 . 2 . M E T H O D S 4 1
3 . 2 . 1 . CELL CULTURE-BASICS 4 1
3.2.1.1. CULTIVATION 4 1
3.2.1.2. PASSAGING 4 1
3.2.1.3. FREEZING AND UNFREEZING 4 1
3.2.1.4. PRECOATING O F DISHES - MATRIX PROTEINS 4 2
3.2.1.5. PRECOATING O F DISHES - FEEDER CELLS 42
3.2.1.5.1. PREPARATION O F MEF CELLS 42
3.2.2. CELL C U L T U R E - M E T H O D S 43
3.2.2.1. HANGING DROP METHOD 43
IMAGE 3
TABLE OF CONTENTS
I I I
3.2.2.2. PRIMARY CELLS 43
3 . 2 . 3 . A N I M A L MODELS OF ISCHEMIC DISEASES 4 4
3.2.3.1. ANIMAL PREPARATION F O R THE OPERATION 44
3.2.3.2. MURINE MODEL O F HINDLIMB ISCHEMIA 44
3.2.3.3. MURINE MODEL O F MYOCARDIAL INFARCTION 45
3.2.3.3.1. BLOOD PERFUSION MEASUREMENT 45
3.2.3.3.2. ISOLATION O F SAMPLES FROM MICE 45
3 . 2 . 4 . I M M U N O C H E M I S T R Y 4 6
3.2.4.1. IMMUNOSTAINING 46
3.2.4.1.1. FACS 47
3.2.4.1.2. MACS 48
3.2.4.2. ELISA 48
3 . 2 . 5 . GENETICS 4 8
3.2.5.1. RNA ISOLATION 48
3.2.5.1.1. RNA QUALITY CONTROL 49
3.2.5.1.2. RNA QUANTIFICATION - NANODROP 49
3.2.5.2. REVERSE TRANSCRIPTION 49
3.2.5.3. PCR - POLYMERASE CHAIN REACTION 50
3.2.5.3.1. QRT-PCR - QUANTITATIVE REAL-TIME-PCR 50
3.2.5.3.2. AGAROSE GEL 5 1
3 . 2 . 6 . IN VITRO ASSAYS 5 1
3.2.6.1. UPTAKE O F LDL 52
3.2.6.2. MATRIGEL ASSAY 52
3.2.6.3. SPHEROID ASSAY 52
3.2.7. IN VIVO ASSAYS 52
3.2.7.1. TUMOUR MODEL 53
3.2.7.2. MATRIGEL PLUG ASSAY 53
4 . R E S U L T S 5 4
4 . 1 . ROLE OF ENDOGENOUS G - C S F IN ISCHEMIA-ASSOCIATED VESSEL
GROWTH 5 4
4.1.1. RECOVERY OF BLOOD PERFUSION IN ISCHEMIC HINDLIMBS 55
4.1.2. ISCHEMIA-ASSOCIATED VESSEL GROWTH 56
4.1.3. INVESTIGATIONS RELATED TO CELLULAR AND MOLECULAR MECHANISMS 58
4.1.3.1. ROLE O F BONE-MARROW-DERIVED CELLS 58
4.1.3.2. CHANGES RELATED T O POTENTIAL MOLECULAR MEDIATORS 60
IMAGE 4
TABLE OF CONTENTS
IV
4.2. GPSCS - POTENTIAL SOURCE OF THERAPEUTICALLY SUITABLE VASCULAR CELLS
6 4
4 . 2 . 1 . DERIVATION OF ENDOTHELIAL CELL-LIKE CELLS F R O M GPSCS 6 4
4 . 2 . 2 . IN VITRO CHARACTERISATION OF GPS-ECS 6 5
4.2.2.1. EXPRESSION O F ENDOTHELIAL SPECIFIC MARKERS ON MRNA-LEVEL 65
4.2.2.2. EXPRESSION O F ENDOTHELIAL SPECIFIC MARKERS ON PROTEIN-LEVEL 66
4.2.2.3. FUNCTIONALITY ASSAYS 67
4.2.3. IN VIVO CHARACTERISATION 68
4.2.3.1. TUMOUR MODEL 69
4.2.3.2. HINDLIMB ISCHEMIA MODEL 70
5 . D I S C U S S I O N 7 1
5 . 1 . ROLE OF ENDOGENOUS G - C S F IN ISCHEMIA-ASSOCIATED VESSEL
GROWTH 7 1
5 . 1 . 1 . RECOVERY OF BLOOD PERFUSION IN ISCHEMIC HINDLIMBS 7 1
5 . 1 . 2 . ISCHEMIA-ASSOCIATED VESSEL GROWTH 7 2
5 . 1 . 3 . INVESTIGATIONS RELATED TO CELLULAR AND MOLECULAR MECHANISMS
7 2
5 . 1 . 3 . 1 . ROLE O F BONE-MARROW-DERIVED CELLS 7 3
5 . 1 . 3 . 2 . CHANGES RELATED T O POTENTIAL MOLECULAR MEDIATORS 7 5
5 . 2 . GPSCS - POTENTIAL SOURCE FOR THERAPEUTICALLY SUITABLE VASCULAR
CELLS 7 6
5 . 2 . 1 . DERIVATION OF ENDOTHELIAL CELL-LIKE CELLS FROM GPSCS 7 6
5 . 2 . 2 . IN VITRO CHARACTERISATION OF GPS-ECS 7 6
5 . 2 . 2 . 1 . EXPRESSION O F ENDOTHELIAL SPECIFIC MARKERS ON
MRNA-LEVEL 7 6
5 . 2 . 2 . 2 . EXPRESSION O F ENDOTHELIAL SPECIFIC MARKERS ON
PROTEIN-LEVEL 7 7
5 . 2 . 2 . 3 . FUNCTIONALITY ASSAYS 7 8
5 . 2 . 3 . IN VIVO CHARACTERISATION 7 8
5 . 2 . 3 . 1 . T U M O U R MODEL 7 8
5 . 2 . 3 . 2 . HINDLIMB ISCHEMIA MODEL 7 9
6 . S U M M A R Y 80
|
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| author | Eligehausen, Sarah Yoon Hee 1983- |
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| dewey-ones | 610 - Medicine and health |
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| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| format | Thesis Book |
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| language | English |
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| spelling | Eligehausen, Sarah Yoon Hee 1983- Verfasser (DE-588)173875580 aut Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies vorgelegt von Sarah Yoon Hee Eligehausen cell-based 2012 Getr. Zählung Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Münster (Westfalen), Univ., Diss., 2012 G-CSF (DE-588)4391233-3 gnd rswk-swf Angiogenese (DE-588)4142449-9 gnd rswk-swf Kardiovaskuläre Krankheit (DE-588)4024666-8 gnd rswk-swf Vascular endothelial Growth Factor (DE-588)4319404-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Kardiovaskuläre Krankheit (DE-588)4024666-8 s Angiogenese (DE-588)4142449-9 s G-CSF (DE-588)4391233-3 s Vascular endothelial Growth Factor (DE-588)4319404-7 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025497233&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Eligehausen, Sarah Yoon Hee 1983- Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies G-CSF (DE-588)4391233-3 gnd Angiogenese (DE-588)4142449-9 gnd Kardiovaskuläre Krankheit (DE-588)4024666-8 gnd Vascular endothelial Growth Factor (DE-588)4319404-7 gnd |
| subject_GND | (DE-588)4391233-3 (DE-588)4142449-9 (DE-588)4024666-8 (DE-588)4319404-7 (DE-588)4113937-9 |
| title | Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies |
| title_alt | cell-based |
| title_auth | Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies |
| title_exact_search | Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies |
| title_full | Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies vorgelegt von Sarah Yoon Hee Eligehausen |
| title_fullStr | Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies vorgelegt von Sarah Yoon Hee Eligehausen |
| title_full_unstemmed | Vascular growth a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies vorgelegt von Sarah Yoon Hee Eligehausen |
| title_short | Vascular growth |
| title_sort | vascular growth a basic study of the role of endogenous g csf and a therapeutic study of its enhancement by cell based therapies |
| title_sub | a basic study of the role of endogenous G-CSF and a therapeutic study of its enhancement by cell based therapies |
| topic | G-CSF (DE-588)4391233-3 gnd Angiogenese (DE-588)4142449-9 gnd Kardiovaskuläre Krankheit (DE-588)4024666-8 gnd Vascular endothelial Growth Factor (DE-588)4319404-7 gnd |
| topic_facet | G-CSF Angiogenese Kardiovaskuläre Krankheit Vascular endothelial Growth Factor Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025497233&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT eligehausensarahyoonhee vasculargrowthabasicstudyoftheroleofendogenousgcsfandatherapeuticstudyofitsenhancementbycellbasedtherapies AT eligehausensarahyoonhee cellbased |