Labeling of hematopoietic stem cells in situ:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English German |
| Veröffentlicht: |
2013
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in engl. und dt. Sprache |
| Beschreibung: | X, 161 S. Ill., graph. Darst. |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV041083364 | ||
| 003 | DE-604 | ||
| 005 | 20130620 | ||
| 007 | t | ||
| 008 | 130612s2013 ad|| m||| 00||| eng d | ||
| 035 | |a (OCoLC)854726287 | ||
| 035 | |a (DE-599)BSZ383027160 | ||
| 040 | |a DE-604 |b ger | ||
| 041 | 0 | |a eng |a ger | |
| 049 | |a DE-188 |a DE-355 |a DE-29T |a DE-83 | ||
| 082 | 0 | |a 573.155 |2 22//ger | |
| 100 | 1 | |a Busch, Katrin |e Verfasser |0 (DE-588)1035439603 |4 aut | |
| 245 | 1 | 0 | |a Labeling of hematopoietic stem cells in situ |c vorgelegt von Katrin Busch |
| 264 | 1 | |c 2013 | |
| 300 | |a X, 161 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Zsfassung in engl. und dt. Sprache | ||
| 502 | |a Heidelberg, Univ., Diss., 2013 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026060103&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-026060103 | ||
Datensatz im Suchindex
| _version_ | 1804150455864918016 |
|---|---|
| adam_text | IMAGE 1
CONTENT
SUMMARY V
Z U S A M M E N F A S S U N G V I
A B B R E V I A T I O N S V I I I
1 I N T R O D U C T I O N 1
1 . 1 HEMATOPOIESIS 1
1 . 1 . 1 HEMATOPOIETIC STEM CELLS 1
1 . 1 . 2 MODELS OF HEMATOPOIESIS 4
1 . 1 . 3 FETAL HEMATOPOIESIS 6
1 . 2 HEMATOPOIETIC STEM CELL ASSAYS 8
1 . 2 . 1 I N VITRO EXPERIMENTS 9
1 . 2 . 2 I N VIVO EXPERIMENTS 9
1 . 2 . 3 LINEAGE TRACING 1 1
1 . 2 . 3 . 1 INDUCIBLE LINEAGE TRACING OF HEMATOPOIETIC LINEAGES 1 2
1 . 2 . 3 . 2 CLONAL TRACKING OF HEMATOPOIETIC STEM CELLS 1 5
2 M A T E R I A L 1 7
2 . 1 CHEMICALS 1 7
2 . 2 ENZYMES 1 8
2 . 3 KITS 1 9
2 . 4 CIRCULAR D N A 1 9
2 . 5 OLIGONUCLEOTIDES 2 0
2 . 6 ANTIBODIES AND ANTISERA 2 3
2 . 7 PLASTICS AND TOOLS 2 6
2 . 8 LABORATORY EQUIPMENT 2 7
2 . 9 SOFTWARE 2 8
2 . 1 0 CELL LINES, BACTERIAL STRAINS A N D MICE 2 8
2 . 1 0 . 1 EUKARYOTIC CELL LINES AND MEDIA 2 8
2 . 1 0 . 2 BACTERIAL STRAINS 2 9
2 . 1 0 . 3 MOUSE STRAINS 3 0
3 M E T H O D S 3 1
3 . 1 MOLECULAR BIOLOGY 3 1
3 . 1 . 1 CLONING - 3 1
I
HTTP://D-NB.INFO/1037137434
IMAGE 2
3.1.1.1 GENERATION OF ELECTROCOMPETENT BACTERIA 32
3.1.1.2 TRANSFORMATION OF PLASMID DNA INTO BACTERIA BY ELECTROPORATION
32
3.1.1.3 GLYCEROL STOCKS O F BACTERIA 3 3
3.1.1.4 ALKALINE LYSIS PLASMID PREPARATION 3 3
3.1.1.5 RESTRICTION ENDONUCLEASE DIGESTION 3 3
3.1.1.6 AGAROSE GEL ELECTROPHORESIS 3 3
3.1.1.7 GEL EXTRACTION O F DNA FRAGMENTS 3 4
3.1.1.8 LIGATION O F DNA FRAGMENTS 3 4
3.1.1.9 PHOSPHORYLATION AND HYBRIDIZATION OF COMPLEMENTARY
OLIGONUCLEOTIDES 3 5
3.1.1.10 SEQUENCING OF DNA 3 5
3.1.2 RNAAND DNA 3 6
3.1.2.1 ISOLATION OF GENOMIC DNA FROM MOUSE TAIL BIOPSIES 3 6
3.1.2.2 ISOLATION OF GENOMIC DNA BY PHENOL/CHLOROFORM EXTRACTION 3 6
3.1.2.3 ISOLATION OF RNA FROM MURINE CELLS O R TISSUES 3 7
3.1.2.4 SYNTHESIS O F CDNA FROM RNA [REVERSE TRANSCRIPTASE-PCR, RT-PCR)
3 7
3.1.2.5 POLYMERASE CHAIN REACTION (PCR) 3 8
3.1.2.5.1 STANDARD PCR 3 9
3.1.2.5.2 PROOF READING PCR 39
3.1.2.5.3 LONG RANGE PCR 4 0
3.1.2.6 SOUTHERN BLOT 4 0
3 . 2 SDS-PAGE AND WESTERN BLOT 4 2
3 . 3 IMMUNOFLUORESCENCE ANALYSIS OF TISSUE SECTIONS 4 3
3 . 4 CELLULAR BIOLOGY 4 4
3 . 4 . 1 MURINE EMBRYONIC FIBROBLASTS - GENERATION AND CULTURE 4 4
3 . 4 . 2 M U R I N E EMBRYONIC STEM CELLS-CULTURE AND GENE TARGETING 4
5
3.4.2.1 MAINTANANCE 4 5
3.4.2.2 TRANSFECTION 4 6
3.4.2.3 SELECTION AND SCREENING 4 6
3.4.2.4 IN VITRO DELETION OF THE NEOMYCIN RESISTANCE GENE 4 7
3.4.2.5 MEF FREE CULTIVATION 4 8
3.4.2.6 EMBROID BODY FORMATION 4 9
3 . 4 . 3 GENERATION OF TRANSGENIC MICE 4 9
3 . 5 FLOWCYTOMETRY 5 0
3 . 5 . 1 ISOLATION O F MURINE CELLS 5 0
3 . 5 . 2 IMMUNOSTAINING 5 0
3 . 5 . 3 LINEAGE DEPLETION 5 1
3 . 5 . 4 CELL SORTING 5 1
II
IMAGE 3
3 . 6 WORKING WITH MICE 5 1
3 . 6 . 1 PREPARATION AND ADMINISTRATION OF TAMOXIFEN 5 2
3 . 6 . 2 B O N E MARROW TRANSPLANTATION 5 2
3.6.3 ISOLATION OF MONONUCLEAR CELLS FROM MURINE PERIPHERAL BLOOD 5 3
4 R E S U L T S 5 4
4 . 1 P A R T I: GENERATION OF MUTANT T I E I AND T I E 2 ALLELES A N D
REPORTER GENE
INDUCTION 5 4
4.1.1 T I E I AND T I E 2 EXPRESSION IN HEMATOPOIETIC CELLS 5 4
4 . 1 . 2 GENERATION OF MUTANT TIE1 MERCREMER/+, TIE2MERCREMER/+ AND
TIE2BCLMER,CREMER/+ E S CELLS
AND MICE 5 7
4.1.2.1 VECTOR DESIGN AND CONSTRUCTION 57
4.1.2.2 GENE TARGETING 59
4.1.2.3 GENERATION OF MUTANT MICE 62
4.1.3 I N VITRO REPORTER GENE INDUCTION IN P A N - M E R C R E M E R R O
S A 2 6 R F P ES CELLS 62
4.1.3.1 GENERATION OF PAN-MERCREMER ROSA26 RFP ES CELLS 62
4.1.3.2 REPORTER GENE INDUCTION IN PAN-MERCREMER ROSA26 RFP ES CELLS 6 3
4 . 1 . 4 I N VIVO REPORTER GENE INDUCTION IN TIE1 MERCREME*/+ AND
TIE2MERCREMER/+ MICE 6 5
4.1.4.1 MERCREMER MRNA EXPRESSION 6 5
4.1.4.2 REPORTER GENE INDUCTION 6 6
4 . 2 P A R T II: CHARACTERIZATION OF T I E 2 8 A M A A C K E M E R / +
MICE 6 9
4 . 2 . 1 EMBRYONIC LETHALITY IN T H E HOMOZYGOUS STATE 6 9
4 . 2 . 2 IMMUNOPHENOTYPICAL CHARACTERIZATION 7 0
4 . 2 . 3 I N VIVO REPORTER GENE INDUCTION 7 1
4.2.4 LIFE SPAN OF MYELOID PRECURSOR CELLS 72
4.2.5 ANALYSIS OF T H E PROPERTIES O F INDIVIDUALLY LABELED HSCS 7 4
4.2.5.1 TRANSPLANTATION OF R/E2^ ,MERICREMER / + LT-HSCS 7 4
4.2.5.2 SINGLE CELL TRANSPLANTATION OF 7 , /E2 MER CREMER A LT-HSCS
77
4 . 2 . 5 . 3 ANALYSIS OF THE COMPETITIVE FITNESS OF TIE2P9 MERTCREMER/+
L T - H S C S 7 8
4 . 2 . 5 . 3 . 1 COMPETITIVE TRANSPLANTATION OF TIE2PA LME CREMER/+ A N
( J ^ L T - H S C S 7 8
4.2.5.3.2 COMPETITIVE TRANSPLANTATION OF YFP + A N D YFP- LT-HSCS FROM
TIE2P9 IMERICREMER/+ROSA26YFP MICE 8 0
4 . 3 P A R T III: LINEAGE TRACING IN TIE2 B :IM CKEME*/+ R OSA26VFP
MICE 8 2
4 . 3 . 1 STABLE LABELING OF LT-HSCS 8 2
4 . 3 . 2 HEMATOPOIETIC OUTPUT O F LABELED LT-HSCS 8 3
4 . 3 . 3 PRECURSOR AND PRODUCT LABELING FREQUENCIES 8 5
4 . 3 . 4 CORRELATION OF LABELED HEMATOPOIETIC CELL SUBSETS 8 8
III
IMAGE 4
4.3.4.1 MEASUREMENT OF LYMPHOID AND MYELOID STEM CELL CONTRIBUTION 8 8
4.3.4.2 CORRELATIONS OF HEMATOPOIETIC PRECURSORS AND PRODUCTS 9 0
4 . 3 . 5 SIMULATION OF STEADY-STATE HEMATOPOIESIS BY A TRANSFORMATION
MATRIX 9 3
4 . 3 . 6 LIMITING DILUTION ASSAYS 9 6
4 . 3 . 7 SPORADIC STEM CELL CONTRIBUTION TO THYMOCYTE DEVELOPMENT 1 0 0
4 . 3 . 8 I N UTERO TAMOXIFEN TREATMENT 1 0 2
4 . 4 PART IV: LINEAGE TRACING IN TIE2 BCLMER,CREME /+ R OSA26^P/RFP M
I C E 1 0 4
4 . 4 . 1 CONFIRMATION OF T H E LABELING PHENOTYPE 1 0 5
4 . 4 . 2 LINEAGE RELATIONSHIPS IN GENETIC MOSAICS 1 0 8
4.4.2.1 MAIN HEMATOPOIETIC PROGENITOR AND MATURE CELL SUBSETS 108
4.4.2.2 T CELL SUBSETS 109
4.4.2.3 B CELL SUBSETS 1 1 1
5 D I S C U S S I O N 1 1 3
5.1 R/2 BGLMR CFIEMFI / + - A NOVEL MOUSE STRAIN FOR INDIVIDUAL
LABELING OF HSCS I N VIVO
1 1 3
5 . 2 NORMAL ADULT STEADY-STATE HEMATOPOIESIS IS A HIGHLY POLYCLONAL
PROCESS 1 1 4
5 . 3 ONE THIRD OF THE L T - H S C POOL IS ACTIVE 1 1 7
5 . 4 L T - H S C S CONSTANTLY DRIVE MYELOPOIESIS AS OPPOSED TO SPORADIC
LYMPHOID
CONTRIBUTION 1 1 8
5 . 5 CLASSICAL OR NON-CLASSICAL MODELS OF HEMATOPOIESIS? 1 2 0
5 . 6 HIGHLIGHTING LINEAGE RELATIONSHIPS WITHIN BRANCHES 1 2 3
5 . 7 CRE INDUCERS AND REPORTERS - FINDING THE RIGHT PARTNERS 1 2 6
5 . 8 LIMITATIONS 1 2 7
5 . 9 PERSPECTIVES 1 2 8
6 R E F E R E N C E S 1 3 0
7 S U P P L E M E N T 1 4 7
7 . 1 GENERATION OF GENE TARGETING VECTORS 1 4 7
7 . 2 IMMUNOPHENOTYPES OF HEMATOPOIETIC CELLS 1 5 0
7 . 3 DATA OF ANALYZED MICE 1 5 1
7 . 4 STATISTICAL METHODOLOGY TO DEVELOP THE TRANSFORMATION MATRIX 1 5 4
7 . 5 LIMITING DILUTION ASSAY 1 5 7
A C K N O W L E D G E M E N T S 1 6 1
I V
|
| any_adam_object | 1 |
| author | Busch, Katrin |
| author_GND | (DE-588)1035439603 |
| author_facet | Busch, Katrin |
| author_role | aut |
| author_sort | Busch, Katrin |
| author_variant | k b kb |
| building | Verbundindex |
| bvnumber | BV041083364 |
| ctrlnum | (OCoLC)854726287 (DE-599)BSZ383027160 |
| dewey-full | 573.155 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 573 - Specific physiological systems in animals |
| dewey-raw | 573.155 |
| dewey-search | 573.155 |
| dewey-sort | 3573.155 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01142nam a2200301 c 4500</leader><controlfield tag="001">BV041083364</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">20130620 </controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">130612s2013 ad|| m||| 00||| eng d</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)854726287</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BSZ383027160</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield><subfield code="a">ger</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-188</subfield><subfield code="a">DE-355</subfield><subfield code="a">DE-29T</subfield><subfield code="a">DE-83</subfield></datafield><datafield tag="082" ind1="0" ind2=" "><subfield code="a">573.155</subfield><subfield code="2">22//ger</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Busch, Katrin</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)1035439603</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Labeling of hematopoietic stem cells in situ</subfield><subfield code="c">vorgelegt von Katrin Busch</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2013</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">X, 161 S.</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="500" ind1=" " ind2=" "><subfield code="a">Zsfassung in engl. und dt. Sprache</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="a">Heidelberg, Univ., Diss., 2013</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026060103&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-026060103</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV041083364 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T00:39:12Z |
| institution | BVB |
| language | English German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-026060103 |
| oclc_num | 854726287 |
| open_access_boolean | |
| owner | DE-188 DE-355 DE-BY-UBR DE-29T DE-83 |
| owner_facet | DE-188 DE-355 DE-BY-UBR DE-29T DE-83 |
| physical | X, 161 S. Ill., graph. Darst. |
| publishDate | 2013 |
| publishDateSearch | 2013 |
| publishDateSort | 2013 |
| record_format | marc |
| spelling | Busch, Katrin Verfasser (DE-588)1035439603 aut Labeling of hematopoietic stem cells in situ vorgelegt von Katrin Busch 2013 X, 161 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in engl. und dt. Sprache Heidelberg, Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026060103&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Busch, Katrin Labeling of hematopoietic stem cells in situ |
| subject_GND | (DE-588)4113937-9 |
| title | Labeling of hematopoietic stem cells in situ |
| title_auth | Labeling of hematopoietic stem cells in situ |
| title_exact_search | Labeling of hematopoietic stem cells in situ |
| title_full | Labeling of hematopoietic stem cells in situ vorgelegt von Katrin Busch |
| title_fullStr | Labeling of hematopoietic stem cells in situ vorgelegt von Katrin Busch |
| title_full_unstemmed | Labeling of hematopoietic stem cells in situ vorgelegt von Katrin Busch |
| title_short | Labeling of hematopoietic stem cells in situ |
| title_sort | labeling of hematopoietic stem cells in situ |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026060103&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT buschkatrin labelingofhematopoieticstemcellsinsitu |