Verfügbarkeit: Influence of young and aged primary stromal cells on leukemic/stromal interactions

Influence of young and aged primary stromal cells on leukemic/stromal interactions:

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Bibliographische Detailangaben
1. Verfasser:	Kirsten, Nicole (VerfasserIn)
Format:	Abschlussarbeit Buch
Sprache:	English
Veröffentlicht:	Ulm [2018]
Schlagworte:	Akute myeloische Leukämie Leukemia, myeloid, acute. Stromal cells. Hochschulschrift
Online-Zugang:	Inhaltsverzeichnis Inhaltsverzeichnis
Beschreibung:	V, 92 Blätter Illustrationen, Diagramme 30 cm

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adam_text	TABLE OF CONTENTS TABLE OF CONTENTS........................................................................................................I ABBREVIATIONS............................................................................................................. ILL 1 INTRODUCTION............................................................................................................. 1 1.1 HAEMATOPOIESIS.............................................................................................. 1 1.2 INFLUENCE OF MICROENVIRONMENT TO HAEMATOPOIESIS AND HSC FATE ....... 2 1.3 AGING IN HAEMATOPOIESIS..............................................................................5 1.4 MICROENVIRONMENTAL CHANGES UPON AG ING ................................................. 6 1.5 ACUTE MYELOID LEUKAEMIA............................................................................... 7 1.6 IMPACT OF AN AGED MICROENVIRONMENT ON LEUKAEMIA ............................... 9 1.7 AIM OF THIS STUDY........................................................................................... 11 2 MATERIALS................................................................................................................ 12 3 METHODS................................................................................................................. 18 3.1 AMPLIFICATION OF PLASMIDS............................................................................ 18 3.2 PRODUCTION OF RETROVIRAL VECTORS ................................................................. 18 3.3 TRANSDUCTION OF MURINE BONE MARROW CELLS ............................................. 19 3.4 FREEZING OF SUSPENSION AND ADHERENT CELLS.............................................20 3.5 THAWING OF SUSPENSION AND ADHERENT CELLS.............................................20 3.6 TRYPAN BLUE EXCLUSION.................................................................................. 20 3.7 ISOLATION AND PURIFICATION OF PRIMARY MURINE STROMAL CELLS .................. 21 3.8 CO-CULTURING OF PRIMARY STROMAL CELLS WITH LONG-TERM CULTURED LEUKEMIC AND PRE-LEUKEMIC CELLS................................................................. 24 3.9 TERMINATION OF CO-CULTURE FOR DIFFERENT ASSAYS........................................25 3.9.1 COLLECTION OF CELL FREE MEDIA FROM CO-CULTURES FOR LATER CYTOKINE ARRAY..................................................................................25 3.9.2 RETRIEVAL OF PRE-LEUKEMIC/LEUKEMIC LTC CELLS AFTER CO-CULTURING FOR C F C S ......................................................................................................... 25 3.9.3 COLLECTION OF STROMAL CELLS AFTER CO-CULTURING.............................................25 3.9.4 FLOW CYTOMETRIC STAINING FOR STROMAL CELLS ................................................. 26 3.9.5 RECOVERY OF PURE STROMAL AND PRE-LEUKEMIC/LEUKEMIC CELLS VIA CELL SORTING..............................................................................................................26 3.10 RNA SEQUENCING.........................................................................................27 3.11 CYTOKINE ARRAY.............................................................................................28 3.12 COLONY FORMING CELL (CFC) ASSAY.............................................................29 3.13 APOPTOSIS STAINING.................................................................................... 29 3.14 OXIDATIVE STRESS DETECTION WITH CELLROX DEEP RED REAGENT ................ 29 3.15 INTERFERENCE ANALYSIS................................................................................... 30 3.16 SS-GALACTOSIDASE STAINING AND ADIPOCYTE COUNTING OF STROMAL CELLS. 30 3.17 FLOW CYTOMETRIC ANALYSIS OF STROMAL COMPOSITION ............................... 31 3.18 TRANSPLANTATION AND MONITORING OF C57BI/6 X C3H/HEJ MICE .............. 31 3.19 ANALYSIS OF DISEASED MICE.........................................................................32 3.20 MAY-GRUENWALD GIEMSA STAINING ............................................................... 33 3.21 IMMUNOPHENOTYPING OF CELLS EX VIVO ....................................................33 3.22 STATISTICAL ANALYSIS......................................................................................34 4 RESULTS................................................................................................................... 35 4.1 VALIDATION OF THE LEUKEMIC POTENTIAL OF PRE-LEUKEMIC AND LEUKEMIC CELLS................................................................................................ 35 4.2 PURITY AND CELLULAR COMPOSITION OF YOUNG AND AGED PRIMARY STROMAL CELLS..........................................................................40 4.3 TRANSPLANTATION OF LEUKEMIC CELLS INTO A YOUNG OR AGED MICROENVIRONMENT SHOWED LONGER DISEASE LATENCY IN YOUNG M ICE ..... 44 4.4 INVESTIGATION OF COMMUNICATION BETWEEN BONE MARROW STROMAL AND PRE-LEUKEMIC/LEUKEMIC CELLS UPON AGING ........................................... 47 4.5 CHANGES IN CLONOGENIC POTENTIAL BY CO-CULTURING....................................48 4.6 DIFFERENCES IN PROLIFERATION UPON CO-CULTURING OF LTC CELLS ON YOUNG AND AGED STROMAL CELLS............................................................... 49 4.7 LTC CELL VIABILITY............................................................................................51 4.8 OXIDATIVE STRESS LEVELS IN LTC CELLS UPON CO-CULTURING WITH YOUNG OR AGED STROMAL CELLS .................................................................................... 53 4.9 DIVERSE CYTOKINE PROFILES OF LEUKEMIC CO-CULTURES UPON AGING .......... 55 4.10 TRANSCRIPTOME ANALYSIS AFTER CROSSTALK BETWEEN BONE MARROW STROMAL AND LEUKEMIC CELLS UPON AG ING .................................................. 60 5 DISCUSSION............................................................................................................74 6 SUM MARY............................................................................................................... 82 7 REFERENCE LIST...................................................................................................... 84 8 ACKNOWLEDGMENT................................................................................................. 91 9 STATUTORY DECLARATION...........................................................................................92
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author	Kirsten, Nicole
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spelling	Kirsten, Nicole Verfasser (DE-588)1160685916 aut Influence of young and aged primary stromal cells on leukemic/stromal interactions Nicole Kirsten Ulm [2018] V, 92 Blätter Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation International Graduate School in Molecular Medicine Ulm 2018 Akute myeloische Leukämie (DE-588)4245523-6 gnd rswk-swf Leukemia, myeloid, acute. Stromal cells. (DE-588)4113937-9 Hochschulschrift gnd-content Akute myeloische Leukämie (DE-588)4245523-6 s DE-604 Universität Ulm (DE-588)30607-1 dgg B:DE-101 application/pdf http://d-nb.info/1161870709/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030764073&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Kirsten, Nicole Influence of young and aged primary stromal cells on leukemic/stromal interactions Akute myeloische Leukämie (DE-588)4245523-6 gnd
subject_GND	(DE-588)4245523-6 (DE-588)4113937-9
title	Influence of young and aged primary stromal cells on leukemic/stromal interactions
title_auth	Influence of young and aged primary stromal cells on leukemic/stromal interactions
title_exact_search	Influence of young and aged primary stromal cells on leukemic/stromal interactions
title_full	Influence of young and aged primary stromal cells on leukemic/stromal interactions Nicole Kirsten
title_fullStr	Influence of young and aged primary stromal cells on leukemic/stromal interactions Nicole Kirsten
title_full_unstemmed	Influence of young and aged primary stromal cells on leukemic/stromal interactions Nicole Kirsten
title_short	Influence of young and aged primary stromal cells on leukemic/stromal interactions
title_sort	influence of young and aged primary stromal cells on leukemic stromal interactions
topic	Akute myeloische Leukämie (DE-588)4245523-6 gnd
topic_facet	Akute myeloische Leukämie Hochschulschrift
url	http://d-nb.info/1161870709/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030764073&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
work_keys_str_mv	AT kirstennicole influenceofyoungandagedprimarystromalcellsonleukemicstromalinteractions AT universitatulm influenceofyoungandagedprimarystromalcellsonleukemicstromalinteractions

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