Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model: = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Würzburg
2017
|
| Subjects: | |
| Online Access: | Volltext Inhaltsverzeichnis Inhaltsverzeichnis |
| Physical Description: | XI, 103 Blätter Illustrationen, Diagramme 30 cm |
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| 245 | 1 | 0 | |a Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model |b = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell |c submitted by Solange Le Blanc Soto from Santiago, Chile |
| 246 | 1 | 1 | |a Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell |
| 264 | 1 | |a Würzburg |c 2017 | |
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Record in the Search Index
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| adam_text | ROLE OF FGF SIGNALING IN THE ADIPOGENIC AND OSTEOGENIC DIFFERENTIATION
OF
HUM AN BONE M ARROW STROM AL CELLS IN A THREE-DIM ENSIONAL IN VITRO
MODEL.
ROLLE DER FGF-SIGNALGEBUNG BEI DER ADIPOGENEN UND OSTEOGENEN
DIFFERENZIERUNG VON HUM ANEN K NOCHENM ARKSTROM AZELLEN IN EINEM
DREIDIM ENSIONALEN IN VITRO MODELL.
DOCTORAL THESIS FOR A DOCTORAL DEGREE
AT THE GRADUATE SCHOOL OF LIFE SCIENCES,
JULIUS-MAXIMILIANS-UNIVERSITAET WUERZBURG,
SECTION BIOMEDICINE
SUBMITTED BY
SOLANGE LE BLANC SOTO
FROM
SANTIAGO, CHILE
WUERZBURG, 2017
TABLE OF CONTENTS
ACKNOWLEDGMENTS__________________________________________________________
VII
ABSTRACT__________________________________________________________________VIII
ZUSAMMENFASSUNG
__________________________________________________________
X
1. INTRODUCTION
______________________________________________________________
1
1.1. HUMAN BONE MARROW STROMAL CELLS
_____________________________________________
1
1.1.1. BONE AND BONE MARROW ADIPOSE TISSUE
_______________________________________
1
1.1.2. IN VITRO CULTURE AND DIFFERENTIATION OF HBMSCS
_________________________________
3
1.1.2.1. OSTEOGENIC
DIFFERENTIATION_____________________________________________ 4
1.1.2.2. ADIPOGENIC
DIFFERENTIATION_____________________________________________ 5
1.1.2.3. HBMSC DIFFERENTIATION BALANCE
_________________________________________
5
1.2. FIBROBLAST GROWTH FACTOR
SIGNALING______________________________________________6
1.2.1. FGFS AND FGF RECEPTORS
___________________________________________________
6
1.2.2. FGF1
SUBFAMILY__________________________________________________________ 8
1.2.3. PARACRINE FGF SIGNALING AND B O N
E___________________________________________ 8
1.3. THREE-DIMENSIONAL CULTURE MODELS FOR HBMSC
___________________________________
9
1.3.1. COLLAGEN HYDROGELS
______________________________________________________
10
1.3.1.1. HBMSCS AND COLLAGEN MATRIX INTERACTION
________________________________
11
1.4. HYPOTHESIS AND AIMS
_______________________________________________________
12
2. MATERIALS AND
METHODS____________________________________________________ 14
2.1.
MATERIALS_________________________________________________________________
14
2.1.1. REAGENTS AND STOCK SOLUTIONS
______________________________________________
14
2.1.2.
CONSUMABLES___________________________________________________________
16
2.1.3.
EQUIPMENT_____________________________________________________________17
2.1.4.
SOFTWARE_______________________________________________________________
18
2.2.
METHODS__________________________________________________________________18
2.2.1. CELL CULTURE
___________________________________________
*
________________
18
2.2.1.1. ISOLATION OF HBMSCS FROM TRABECULAR
BONE_______________________________ 18
2.2.1.2. SUBCULTURING OF HBMSCS
_____________________________________________
18
2.2.1.3. HBMSC-LADEN 3D COLLAGEN G ELS
________________________________________
19
2.2.1.4. INDUCTION OF ADIPOGENIC
DIFFERENTIATION__________________________________19
2.2.1.5. INDUCTION OF OSTEOGENIC
DIFFERENTIATION__________________________________20
2.2.1.6. TREATMENT WITH RHFGFS AND FGFR INHIBITORS
____________________________
20
2.2.1.7. DEAD CELL STAINING IN COLLAGEN GELS
______________________________________
20
2.2.1.8. DNA
QUANTIFICATION__________________________________________________21
2.2.2. HISTO- AND
CYTOCHEMISTRY_________________________________________________ 21
2.2.2.1. HEMATOXYLIN AND EOSIN STAINING
_______________________________
21
2.2.2.2. OIL RED 0
STAINING_______________________________________________
___
22
2.2.2.3. ALKALINE PHOSPHATASE ACTIVITY
_______
22
2.2.2.4. ALIZARIN RED S STAINING
______________________________________________
22
2.2.2.5. TIMP1 IMMUNOCYTOCHEMICAL STAINING
__________________________________
22
2.2.2.6. PHALLOIDIN STAINING OF ACTIN
CYTOSKELETON________________________________ 23
2.22.1. IMAGE ACQUISITION
_____________________________________
23
2.2.3. MOLECULAR
BIOLOGY_______________________________________________________ 23
2.2.3.1. RNA
ISOLATION______________________________________________________ 23
2.2.3.2. CDNA
SYNTHESIS_____________________________________________________24
2.2.3.3. QUANTITATIVE PGR
_______________________________________
24
2.2.4. STATISTICAL
ANALYSES______________________________________________________ 25
3. RESULTS
_______________________
26
3.1. ESTABLISHMENT AND CHARACTERIZATION OF A 3D CULTURE MODEL FOR
ADIPOGENIC AND OSTEOGENIC
DIFFERENTIATION OF HBMSCS BASED ON COLLAGEN GELS
__
_______________________________
_
26
3.1.1. OPTIMIZATION OF COLLAGEN GEL SETTING CONDITIONS FOR HBMSCS
DIFFERENTIATION________26
3.1.2. HBMSCS DIFFERENTIATION IN 3D COLLAGEN GELS
____
______________________________29
3.1.2.1. ADIPOGENIC AND OSTEOGENIC
DIFFERENTIATION______________________________ 29
3.1.2.2. EXPRESSION OF MATRIX REMODELING M
ARKERS_______________________________ 32
3.1.2.3. 3D COLLAGEN GELS VS MONOLAYER CULTURE
_
________________________________
_
33
3.2. INHIBITORY EFFECT OF FGF1 SIGNALING ON HBMSC DIFFERENTIATION IN 3D
COLLAGEN GELS
______
35
3.2.1. RHFGFL TREATMENT INHIBITS HBMSC ADIPOGENESIS AND OSTEOGENESIS IN
3D COLLAGEN GELS36
3.2.1.1. INHIBITION OF ADIPOGENIC DIFFERENTIATION BY RHFGFL IN 3D
COLLAGEN GELS________36
3.2.1.2. INHIBITION OF OSTEOGENIC DIFFERENTIATION BY RHFGFL IN 3D
COLLAGEN GELS________39
3.2.1.3. DNA CONTENT AFTER HBMSC DIFFERENTIATION INHIBITION BY RHFGFL IN
3D COLLAGEN GELS
________________________________________________
40
3.2.2. IMPACT OF FGFR SIGNALING INHIBITION DURING HBMSC DIFFERENTIATION
INDUCTION IN 3D
COLLAGEN
GELS________________________________________________________________ 41
3.2.2.1. HBMSC ADIPOGENIC DIFFERENTIATION IN 3D COLLAGEN GELS UNDER FGFR
SIGNALING
INHIBITION
____________________
41
3.2.2.2. HBMSC OSTEOGENIC DIFFERENTIATION IN 3D COLLAGEN GELS UNDER FGFR
SIGNALING
INHIBITION^_________________________ 43
3.2.3. MATRIX REMODELING IS AFFECTED BY RHFGFL TREATMENT DURING HBMSC
DIFFERENTIATION
INDUCTION IN 3D COLLAGEN GELS
______________
44
3.3. EXPANSION OF HBMSCS IN THE PRESENCE OF
RHFGFL_________________________________ 46
3.3.1. EFFECT OF RHFGFL ON EXPANDING HBMSCS
_____________________________________
47
3.3.1.1. HBMSC PROLIFERATION
_______
47
3.3.1.2. COLLAGEN MATRIX CONTRACTION
_
_________________________________________ 48
3.3.2. DIFFERENTIATION POTENTIAL OF HBMSCS IN 3D COLLAGEN GELS AFTER
EXPANSION WITH RHFGFL 49
3.3.2.1. ADIPOGENIC POTENTIAL
_____
49
3.3.2.2. OSTEOGENIC
POTENTIAL_______________________________________________ 50
4. DISCUSSION
______________________________________________________________
53
4.1. DIFFERENTIATION OF HBMSCS IN 3D COLLAGEN
GELS___________________________________53
4.1.1. 3D COLLAGEN GELS AND HBMSC DIFFERENTIATION
_________________________________
53
4.1.2. HBMSC DIFFERENTIATION AND COLLAGEN MATRIX REMODELING
________________________
55
4.1.2.1. COLLAGEN MATRIX CONTRACTION BY DIFFERENTIATING HBMSCS
___________________
55
4.1.2.2. ROLE OF MATRIX METALLOPROTEINASES AND THEIR INHIBITORS IN
COLLAGEN MATRIX
REMODELING_______________________________________________________________
57
4.2. FGF SIGNALING IN HBMSC DIFFERENTIATION
________________________________________
58
4.2.1. ROLE OF FGF1 AND FGF2 IN ADIPOGENIC
DIFFERENTIATION__________________________ 60
4.2.2. ROLE OF FGF1 AND FGF2 IN OSTEOGENIC DIFFERENTIATION
__________________________
61
4.2.3. INHIBITION OF DIFFERENTIATION VERSUS ENHANCED PROLIFERATION IN
COLLAGEN GELS________64
4.2.4. INHIBITION OF FGFR SIGNALING AND HBMSC DIFFERENTIATION
________________________
64
4.3. FGF SIGNALING DURING HBMSC EXPANSION AND DIFFERENTIATION POTENTIAL
________________
66
4.3.1. FGF-INDUCED PROLIFERATION AND HBMSC
POTENCY_______________________________ 66
4.3.2. FGF SIGNALING AND ADIPOGENIC POTENTIAL
_____________________________________
67
4.3.3. FGF SIGNALING AND OSTEOGENIC POTENTIAL
______________________________________
68
4.4. FGF SIGNALING AND HBMSC MATRIX REMODELING ABILITY
______________________________
70
5. CONCLUSION
______________________________________________________________
74
6. REFERENCES
______________________________________________________________
76
LIST OF
FIGURES______________________________________________________________94
LIST OF TABLES
______________________________________________________________
96
LIST OF
ABBREVIATIONS________________________________________________________ 97
LIST OF PUBLICATIONS
________________________________________________________
100
CURRICULUM
VITAE__________________________________________________________ 101
|
| any_adam_object | 1 |
| author | Le Blanc, Solange 1982- |
| author_GND | (DE-588)1133221238 |
| author_facet | Le Blanc, Solange 1982- |
| author_role | aut |
| author_sort | Le Blanc, Solange 1982- |
| author_variant | b s l bs bsl |
| building | Verbundindex |
| bvnumber | BV044339874 |
| collection | ebook |
| ctrlnum | (OCoLC)1004519901 (DE-599)DNB1132730260 |
| dewey-full | 610 |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 610 - Medicine and health |
| dewey-raw | 610 |
| dewey-search | 610 |
| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| format | Thesis Book |
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| publishDateSort | 2017 |
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| spelling | Le Blanc, Solange 1982- Verfasser (DE-588)1133221238 aut Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell submitted by Solange Le Blanc Soto from Santiago, Chile Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell Würzburg 2017 XI, 103 Blätter Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Julius-Maximilians-Universität Würzburg 2017 Zusammenfassung in deutscher und englischer Sprache Knochenmarkzelle (DE-588)4123534-4 gnd rswk-swf Knochenbildung (DE-588)4031317-7 gnd rswk-swf Fibroblastenwachstumsfaktor (DE-588)4310092-2 gnd rswk-swf Fettzelle (DE-588)4154243-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Fettzelle (DE-588)4154243-5 s Knochenbildung (DE-588)4031317-7 s Knochenmarkzelle (DE-588)4123534-4 s Fibroblastenwachstumsfaktor (DE-588)4310092-2 s DE-604 Erscheint auch als Online-Ausgabe, PDF urn:nbn:de:bvb:20-opus-147659 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-147659 Resolving-System kostenfrei Volltext B:DE-101 application/pdf http://d-nb.info/1132730260/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029742932&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Le Blanc, Solange 1982- Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell Knochenmarkzelle (DE-588)4123534-4 gnd Knochenbildung (DE-588)4031317-7 gnd Fibroblastenwachstumsfaktor (DE-588)4310092-2 gnd Fettzelle (DE-588)4154243-5 gnd |
| subject_GND | (DE-588)4123534-4 (DE-588)4031317-7 (DE-588)4310092-2 (DE-588)4154243-5 (DE-588)4113937-9 |
| title | Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell |
| title_alt | Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell |
| title_auth | Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell |
| title_exact_search | Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell |
| title_full | Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell submitted by Solange Le Blanc Soto from Santiago, Chile |
| title_fullStr | Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell submitted by Solange Le Blanc Soto from Santiago, Chile |
| title_full_unstemmed | Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell submitted by Solange Le Blanc Soto from Santiago, Chile |
| title_short | Role of FGF signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three-dimensional in vitro model |
| title_sort | role of fgf signaling in the adipogenic and osteogenic differentiation of human bone marrow stromal cells in a three dimensional in vitro model rolle der fgf signalgebung bei der adipogenen und osteogenen differenzierung von humanen knochenmarkstromazellen in einem dreidimensionalen in vitro modell |
| title_sub | = Rolle der FGF-Signalgebung bei der adipogenen und osteogenen Differenzierung von humanen Knochenmarkstromazellen in einem dreidimensionalen in vitro Modell |
| topic | Knochenmarkzelle (DE-588)4123534-4 gnd Knochenbildung (DE-588)4031317-7 gnd Fibroblastenwachstumsfaktor (DE-588)4310092-2 gnd Fettzelle (DE-588)4154243-5 gnd |
| topic_facet | Knochenmarkzelle Knochenbildung Fibroblastenwachstumsfaktor Fettzelle Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-147659 http://d-nb.info/1132730260/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029742932&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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