An investigation into the characteristics and potential therapeutic application of human bone marrow-derived mesenchymal stromal cells in experimental spinal cord injury [Elektronische Ressource] / Katrin Montzka

Sujets

• Gesundheit

Informations

• Publié par: rheinisch-westfalischen_technischen_hochschule_-rwth-_aachen
• Publié le: 01 janvier 2011
• Nombre de lectures: 4
• Langue: English
• Poids de l'ouvrage: 12 Mo

Extrait

"An investigation into
the characteristics and potential therapeutic application of
human bone marrow-derived mesenchymal stromal cells
in experimental spinal cord injury"


Von der Fakultät für Mathematik, Informatik und Naturwissenschaften der RWTH
Aachen University zur Erlangung des akademischen Grades einer Doktorin der
Naturwissenschaften genehmigte Dissertation

vorgelegt von


Diplom)Bioingenieurin (FH)
Katrin Montzka

aus Düren


Berichter: Universitätsprofessor Dr. Hermann Wagner
PD Dr. Gary Brook


Tag der mündlichen Prüfung: 09.08.2011


Diese Dissertation ist auf den Internetseiten der Hochschulbibliothek online verfügbar.











































“Every great scientific truth goes through three stages.
First, people say it conflicts with the Bible.
Next they say it has been discovered before.
Lastly they say they always believed it.”

Louis Agassiz (1807-1873)



TABLE OF CONTENTS I
Table of contents
PAGE
1. SUMMARY.................................................................................................................................................. 1
2. ZUSAMMENFASSUNG............................................................................................................................. 3
3. INTRODUCTION....................................................................................................................................... 5
3.1 DEFINITION AND TYPES OF STEM CELLS................................................................................................. 5
3.2 MESENCHYMAL STROMAL CELLS .......................................................................................................... 7
3.3 THERAPEUTIC POTENTIAL OF MESENCHYMAL STROMAL CELLS IN SPINAL CORD INJURY ....................... 8
3.4 HYPOTHESIS AND GOALS OF THE THESIS ............................................................................................. 12
4. NEURAL DIFFERENTIATION POTENTIAL OF HUMAN BONE MARROW-DERIVED
MESENCHYMAL STROMAL CELLS: MISLEADING MARKER GENE EXPRESSION .................... 13
4.1 ABSTRACT........................................................................................................................................... 13
4.2 INTRODUCTION.................................................................................................................................... 13
4.3 MATERIALS AND METHODS ................................................................................................................. 14
4.3.1 Sampling of human MSC................................................................................................................ 14
4.3.2 Isolation and cultivation of human MSC........................................................................................ 14
4.3.3 Characterization of isolated human MSC ...................................................................................... 15
4.3.4 Gene expression analysis ............................................................................................................... 16
4.3.5 Immunocytochemistry..................................................................................................................... 17
4.4 RESULTS.............................................................................................................................................. 17
4.4.1 Characterization of human MSC.................................................................................................... 17
4.4.2 Neural marker expression by undifferentiated human MSC .......................................................... 19
4.4.3 Immunocytochemical detection of neural marker proteins by undifferentiated human MSC ........ 22
4.5 DISCUSSION......................................................................................................................................... 24
4.6 CONCLUSION....................................................................................................................................... 26
5. EXPANSION OF HUMAN BONE MARROW-DERIVED MESENCHYMAL STROMAL CELLS:
SERUM-REDUCED MEDIUM IS BETTER THAN CONVENTIONAL MEDIUM ................................. 27
5.1 ABSTRACT........................................................................................................................................... 27
5.2 INTRODUCTION.................................................................................................................................... 27
5.3 MATERIALS AND METHODS ................................................................................................................. 28
5.3.1 Cell culture..................................................................................................................................... 28
5.3.2 Proliferation and cytotoxicity......................................................................................................... 30
5.3.3 Statistical analysis.......................................................................................................................... 30
5.4 RESULTS.............................................................................................................................................. 30
5.5 DISCUSSION......................................................................................................................................... 33
5.6 CONCLUSION....................................................................................................................................... 34
TABLE OF CONTENTS II
6. GROWTH FACTOR AND CYTOKINE EXPRESSION OF HUMAN MESENCHYMAL
STROMAL CELLS IS NOT ALTERED IN AN IN VITRO MODEL OF TISSUE DAMAGE ................. 35
6.1 ABSTRACT........................................................................................................................................... 35
6.2 INTRODUCTION.................................................................................................................................... 35
6.3 MATERIAL AND METHODS ................................................................................................................... 37
6.3.1 Cell culture..................................................................................................................................... 37
6.3.2 TLR stimulation.............................................................................................................................. 37
6.3.3 Immunocytochemistry..................................................................................................................... 37
6.3.4 Surgical procedure and tissue extraction....................................................................................... 38
6.3.5 Proliferation assay ......................................................................................................................... 39
6.3.6 Cell migration assay ...................................................................................................................... 39
6.3.7 RNA isolation and gene expression analysis.................................................................................. 39
6.3.8 Statistical analysis.......................................................................................................................... 40
6.4 RESULTS.............................................................................................................................................. 40
6.4.1 Characterization of hMSC ............................................................................................................. 40
6.4.2 LPS treatment of hMSC leads to an altered expression of cytokines and growth factors .............. 41
6.4.3 hMSC exposure to tissue homogenates does not alter cytokine or growth factor expression
patterns 42
6.4.4 Stimulation with tissue homogenate triggers hMSC proliferation but not migration .................... 43
6.5 DISCUSSION......................................................................................................................................... 43
6.6 CONCLUSION....................................................................................................................................... 46
7. MESENCHYMAL STROMAL CELLS IMMOBILIZED IN AN ORIENTED 3D SCAFFOLD
TRANSIENTLY ACCELERATE FUNCTIONAL RECOVERY AFTER ACUTE SPINAL CORD
INJURY .............................................................................................................................................................. 48
7.1 ABSTRACT........................................................................................................................................... 48
7.2 INTRODUCTION.................................................................................................................................... 48
7.3 MATERIALS AND METHODS .................................................................................................................