Development of advanced human intestinal in vitro models: = Entwicklung von erweiterten humanen intestinalen in vitro Modellen
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|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Würzburg
2016
|
| Subjects: | |
| Online Access: | Volltext Inhaltsverzeichnis |
| Physical Description: | XV, 121 Blätter Illustrationen, Diagramme |
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| 100 | 1 | |a Schweinlin, Matthias Oliver |d 1985- |e Verfasser |0 (DE-588)1124010114 |4 aut | |
| 245 | 1 | 0 | |a Development of advanced human intestinal in vitro models |b = Entwicklung von erweiterten humanen intestinalen in vitro Modellen |c submitted by Matthias Oliver Schweinlin from Lörrach |
| 246 | 1 | 1 | |a Entwicklung von erweiterten humanen intestinalen in vitro Modellen |
| 264 | 1 | |a Würzburg |c 2016 | |
| 300 | |a XV, 121 Blätter |b Illustrationen, Diagramme | ||
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Record in the Search Index
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| adam_text |
DEVELOPMENT OF ADVANCED HUMAN INTESTINAL
IN VITRO
MODELS
* * *
ENTWICKLUNG VON ERWEITERTEN HUMANEN INTESTINALEN
IN VITRO
MODELLEN
DOCTORAL THESIS FOR A DOCTORAL DEGREE
AT THE GRADUATE SCHOOL OF LIFE SCIENCES,
JULIUS-MAXIMILIANS-UNIVERSITAET WUERZBURG,
SECTION CLINICAL SCIENCES
SUBMITTED BY
MATTHIAS OLIVER SCHWEINLIN
FROM LOERRACH
WUERZBURG 2016
TABLE O F CON TEN TS
LIST OF
FIGURES.
IV
LIST OF
TABLES.
VI
ABBREVIATIONS.
VII
SUMMARY.
XI
ZUSAMMENFASSUNG.XIII
1
INTRODUCTION.
1
1.1 THE SMALL
INTESTINE.1
1.1.1 ANATOMY AND PHYSIOLOGY OF THE SMALL
INTESTINE.1
1.1.2 THE INTESTINAL MUCOSA - CELL TYPES AND MOLECULAR REGULATION
.
2
1.1.3 GUT HOMEOSTASIS -
REGULATION/DYSREGULATION.7
1.2 THE GUT BARRIER - INTESTINAL TRANSPORT MECHANISMS
.
8
1.2.1 PARACELLULAR
TRANSPORT.9
1.2.2 TRANSCELLULAR PASSIVE/ACTIVE
TRANSPORT. 9
1.2.3 EFFLUX
TRANSPORT.
10
1.2.4
ENDOCYTOSIS.
10
1.3 PRECLINICAL MODEL
SYSTEMS.11
1.3.1 IN
VIVO.
11
1.3.2 EX
VIVO.
11
1.3.3 IN
VITRO.
12
1.3.4 IN SILICO MODELLING
.
16
1.4 TISSUE ENGINEERING
.
17
1.5 APPLICATIONS OF PERORAL TEST SYSTEMS IN BIOMEDICAL
RESEARCH.18
2 SCOPE OF
THESIS.20
3
MATERIALS.
21
3.1 BIOLOGICAL
MATERIALS.21
3.2
EQUIPMENT.21
3.3 DISPOSABLE
MATERIALS.
23
3.4 LABORATORY
MATERIALS.24
3.5 CHEMICALS AND
SOLUTIONS.25
3.6 GENERAL BUFFERS AND SOLUTIONS FOR CELL
CULTURE.30
3.7 CELL CULTURE
MEDIUM.
30
3.8 MATERIALS AND SOLUTIONS FOR HISTOLOGY
.
32
3.9
ENZYMES.
33
3.10
ANTIBODIES.
33
3.11
QPCR.
34
3.12
SOFTWARE.
34
4
METHODS.
36
4.1 CELL CULTURE
.
36
4.1.1 PREPARATION OF PORCINE DECELLULARIZED
SCAFFOLDS.36
4.1.2 CACO-2 CELL
LINE.37
4.1.3 DENDRITIC CELLS
.
38
4.1.4 PRIMARY CELLS FROM HUMAN INTESTINAL TISSUE - ISOLATION AND
EXPANSION. 39
4.1.5 WNT3A-CONDITIONED
MEDIUM.40
4.1.6 PRIMARY INTESTINAL TISSUE MODEL - DIFFERENT CULTURE CONDITIONS
.
40
4.2
HISTOLOGY.
41
4.2.1 FIXATION AND EMBEDDING OF TISSUES
.
41
4.2.2 DEPARAFFINIZATION AND REHYDRATION OF TISSUE SECTIONS
.
42
4.2.3 HAEMATOXYLIN & EOSIN (H&E)
STAINING.43
4.2.4 FEULGEN
STAINING.
44
4.2.5 MASSON'S TRICHROME
STAINING.44
4.2.6
IMMUNOFLUORESCENCE.
45
4.2.7 SEM AND TEM
IMAGING.
46
4.3 FLOW
CYTOMETRY.
46
4.4 GENE EXPRESSION ANALYSIS BY QPCR
.
47
4.5 FITC-DEXTRAN
ASSAY.47
4.6
TEER-MEASUREMENT.
48
4.7 TRANSPORT
STUDIES.49
4.8
NANOPARTICLES.
49
5
RESULTS.51
5.1 FUNCTIONAL TESTS ON AN IMPROVED CACO-2
MODEL.51
5.1.1 ADAPTION OF THE CACO-2
MODEL. 51
5.1.2 IMMUNE CELL
DIFFERENTIATION.
52
5.1.3 CACO-2 TRANSPORT ASSAY PERFORMANCE
.
54
5.2 ESTABLISHMENT OF A PRIMARY SMALL INTESTINAL TISSUE MODEL
.
61
5.2.1 PILOT STUDIES FOR MODEL
ESTABLISHMENT.61
5.2.2 ADAPTION AND DETAILED CHARACTERIZATION OF THE PRIMARY MODEL
.
68
5.2.3 INFLUENCE OF CONTROLLED DYNAMIC CULTURE
CONDITIONS.70
5.2.4 MOLECULAR
CHARACTERIZATION.
74
5.2.5 PARTICLE STUDIES ON THE PRIMARY
MODEL.76
6
DISCUSSION.
80
6.1 IMPROVEMENT OF THE CACO-2 MODEL
.
81
6.1.1 DEVELOPMENT OF A CO-CULTURE
MODEL.82
6.1.2 TESTED
FORMULATIONS.
84
6.2 PRIMARY SMALL INTESTINAL TISSUE
MODEL.87
6.2.1 ESTABLISHMENT OF A PRIMARY INTESTINAL MONOLAYER
MODEL.87
6.2.2 CHARACTERISATION OF THE BARRIER
PROPERTIES.90
6.2.3 INFLUENCE OF CONTROLLED DYNAMIC CULTURE CONDITIONS
.
92
6.2.4 MOLECULAR
CHARACTERIZATION.93
6.2.5 PARTICLE STUDIES ON THE PRIMARY
MODEL.94
7 CONCLUSIONS AND FUTURE PERSPECTIVES
.
96
8
REFERENCES.
98
9
APPENDIX.112
9.1 SUPPLEMENTARY
DATA.
112
9.2 CURRICULUM
VITAE.114
9.3 PUBLICATIONS AND CONFERENCE CONTRIBUTIONS
.
116
9.4
DANKSAGUNG.118
9.5 AFFIDAVIT
.
120
9.6 EIDESSTAATLICHE
ERKLAERUNG.121 |
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| spelling | Schweinlin, Matthias Oliver 1985- Verfasser (DE-588)1124010114 aut Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen submitted by Matthias Oliver Schweinlin from Lörrach Entwicklung von erweiterten humanen intestinalen in vitro Modellen Würzburg 2016 XV, 121 Blätter Illustrationen, Diagramme txt rdacontent n rdamedia nc rdacarrier Dissertation Julius-Maximilians-Universität Würzburg 2016 Zusammenfassung in deutscher und englischer Sprache In vitro (DE-588)4250701-7 gnd rswk-swf Tissue Engineering (DE-588)4646061-5 gnd rswk-swf Dünndarm (DE-588)4013218-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Dünndarm (DE-588)4013218-3 s In vitro (DE-588)4250701-7 s Tissue Engineering (DE-588)4646061-5 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-142571 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-142571 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029417926&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schweinlin, Matthias Oliver 1985- Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen In vitro (DE-588)4250701-7 gnd Tissue Engineering (DE-588)4646061-5 gnd Dünndarm (DE-588)4013218-3 gnd |
| subject_GND | (DE-588)4250701-7 (DE-588)4646061-5 (DE-588)4013218-3 (DE-588)4113937-9 |
| title | Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen |
| title_alt | Entwicklung von erweiterten humanen intestinalen in vitro Modellen |
| title_auth | Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen |
| title_exact_search | Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen |
| title_full | Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen submitted by Matthias Oliver Schweinlin from Lörrach |
| title_fullStr | Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen submitted by Matthias Oliver Schweinlin from Lörrach |
| title_full_unstemmed | Development of advanced human intestinal in vitro models = Entwicklung von erweiterten humanen intestinalen in vitro Modellen submitted by Matthias Oliver Schweinlin from Lörrach |
| title_short | Development of advanced human intestinal in vitro models |
| title_sort | development of advanced human intestinal in vitro models entwicklung von erweiterten humanen intestinalen in vitro modellen |
| title_sub | = Entwicklung von erweiterten humanen intestinalen in vitro Modellen |
| topic | In vitro (DE-588)4250701-7 gnd Tissue Engineering (DE-588)4646061-5 gnd Dünndarm (DE-588)4013218-3 gnd |
| topic_facet | In vitro Tissue Engineering Dünndarm Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-142571 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029417926&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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