Atlas of human pluripotent stem cells: derivation and culturing
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
New York, NY [u.a.]
Humana Press, Springer
2012
|
| Schriftenreihe: | Stem cell biology and regenerative medicine
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Klappentext |
| Beschreibung: | XIV, 142 S. zahlr. Ill. |
| ISBN: | 9781617795473 |
Internformat
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| 245 | 1 | 0 | |a Atlas of human pluripotent stem cells |b derivation and culturing |c Michal Amit ; Joseph Itskovitz-Eldor [ed.] |
| 264 | 1 | |a New York, NY [u.a.] |b Humana Press, Springer |c 2012 | |
| 300 | |a XIV, 142 S. |b zahlr. Ill. | ||
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Datensatz im Suchindex
| _version_ | 1804149082520813568 |
|---|---|
| adam_text | Contents
1
Methods for the Derivation of Human Embryonic
Stem Cell Lines
.......................................................................................... 1
1.1
Introduction
........................................................................................ 1
1.2
Materials for
ESC
Line Derivation
.................................................... 9
1.3
Methods for hESC Isolation
.............................................................. 9
1.3.1
hESC Isolation by Immunosurgery
........................................ 12
1.3.2
Mechanical Removal of Trophectoderm
................................ 12
1.3.3
Whole Embryo Approach for ESC Line Derivation
.............. 13
References
................................................................................................... 13
2
Morphology of Human Embryonic and Induced Pluripotent
Stem Cell Colonies Cultured with Feeders
............................................. 15
2.1
Introduction
........................................................................................ 15
2.2
Materials
............................................................................................ 16
2.2.1
For Mouse Embryonic
Fibroblasts
(MEFs)
and Foreskin
Fibroblasts
(HFFs)
........................................... 16
2.2.2
ForhPSC Maintenance
.......................................................... 17
2.3
Methods
............................................................................................. 18
2.3.1
Feeder Culture Methods
........................................................ 18
2.3.2
hPSC Culture
......................................................................... 22
References
................................................................................................... 38
3
Morphology of Human Embryonic Stem Cells and Induced
Pluripotent Stem Cells Cultured in Feeder
Layer-Free Conditions
.............................................................................. 41
3.1
Introduction
........................................................................................ 41
3.2
Materials for Feeder Layer-Free Culture of hPSCs
........................... 43
3.2.1
Matrix Preparation
................................................................. 43
3.2.2
Culture Medium
..................................................................... 44
Contents
3.3
Methods for hPSC Feeder Layer-Free Culture
.................................. 44
3.3.1
Preparation of Matrix-Covered Plates
................................... 44
3.3.2
Splitting, Freezing, and Thawing hPSCs
............................... 45
3.3.3
Adaptation of PSCs to Feeder-Free Culture
.......................... 45
3.3.4
Routine Culture of hPSCs
...................................................... 46
References
................................................................................................... 54
Morphology of Undifferentiated Human Embryonic
and Induced Stem Cells Grown in Suspension
and in Dynamic Cultures
.......................................................................... 57
4.1
Introduction
........................................................................................ 57
4.2
Materials for Suspension Culture of hPSCs
...................................... 58
4.2.1
Culture Medium
..................................................................... 58
4.2.2
Splitting Medium
................................................................... 59
4.2.3
Freezing Medium
................................................................... 59
4.3
Methods for Suspension Culture of hPSCs
....................................... 60
4.3.1
Creating a hPSC Suspension Culture
.................................... 60
4.3.2
Splitting hPSCs in Suspension
............................................... 60
4.3.3
Freezing hPSCs in Suspension
.............................................. 62
4.3.4
Thawing hPSCs in Suspension
.............................................. 64
4.3.5
Culturing hPSCs in a Dynamic System
................................. 64
4.3.6
Routine Culture of hPSCs in Suspension
.............................. 65
References
................................................................................................... 71
Differentiation of Pluripotent Stem Cells In Vitro:
Embryoid Bodies
....................................................................................... 73
5.1
Introduction
........................................................................................ 73
5.2
Materials for EB Formation
............................................................... 75
5.2.1
Culture Medium Supplemented with Serum
......................... 75
5.2.2
Splitting Medium Based on
Collagenase............................... 76
5.3
Methods for EB Formation and Culture
............................................ 76
5.3.1
EB Formation
......................................................................... 76
5.3.2
Routine Culture of EBs
.......................................................... 76
5.3.3
Culturing EBs in Spinner Flasks
............................................ 78
References
................................................................................................... 88
Differentiation of Pluripotent Stem Cells In Vivo:
Teratoma Formation
................................................................................. 91
6.1
Introduction
........................................................................................ 91
6.2
Materials for Teratoma Formation
..................................................... 93
6.2.1
Culture Medium
..................................................................... 93
6.2.2
Syringe for Injecting Cells
..................................................... 93
6.3
Formation of Teratomas
..................................................................... 93
6.3.
1 Protocol for Teratoma Formation
.......................................... 93
6.3.2
Routine Treatment of Mice and Teratoma
............................. 93
References
................................................................................................... 103
Contents
x¡
7 Immunostaining........................................................................................ 105
7.1
Introduction
........................................................................................ 105
7.2 Materials and Solutions
for
Immunostaining.................................... 11
¡
7.2.1 Materials and Solutions
for
Immunohistochemistry
of Paraffin-Embedded Tissues
...............................................
1
11
7.2.2
Materials and Solutions for Immunofluoresccnce
................. 111
7.3
Immunostaining Procedures
..............................................................
і
12
7.3.1
Immunohistochemistry of Paraffin-Embedded Tissues
......... 112
7.3.2
Immunofluorescence of Cultured Cells
................................. 113
References
...................................................................................................
1
13
8
Karyotype and Fluorescent In Situ Hybridization
Analysis of Human Embryonic Stem Cell and Induced
Pluripotent Stem Cell Lines
..................................................................... 115
8.1
Introduction
........................................................................................ 115
8.1.1
Karyotype Analysis
............................................................... 115
8.1.2
FISH Analysis
........................................................................ 121
8.2
Materials for Harvesting Cells for Karyotyping
and FISH Analysis
.............................................................................
1
24
8.2.1
Reagents
................................................................................. 124
8.2.2
Solutions
................................................................................ 124
8.3
Procedure of Harvesting Cells for Karyotyping
and FISH Analysis
......
ľ.
...................... ..............................................
1
24
References
...................................................................................................
12ft
9
Method for the Derivation of Induced Pluripotent
Stem Cells from Human Hair Follicle Keratinocytes
............................ ) 27
9.1
Introduction
........................................................................................ 127
9.2
Materials
............................................................................................ 129
9.2.1
NIH-3T3/293T Cells
.............................................................. 129
9.2.2
Keratinocyte Derivation from Plucked Hair Follicles
........... 129
9.3
Methods
............................................................................................. 130
9.3.1
NIH-3T3 and 293T Culture Methods
.................................... 130
9.3.2
Keratinocyte Culture Methods
............................................... 132
9.3.3
Preparation of the STEMCCA Virus for Infection
................ 133
9.3.4
Derivation of iPSCs from Hair Keratinocytes
....................... 134
References
................................................................................................... 137
About the Authors
........................................................................................... 139
Index
................................................................................................................. 141
Stem Cell Biology and Regenerative Medicine
Kursad Turksen, Series Editor
Michal
Amit
■
Joseph Itskovitz-Eldor Editors
Atlas of Human Pluripotent Stem Cells
Derivation and Culturing
Human pluripotent stem cells, including human embryonic stem cells and induced
pluripotent stem cells, are a key focus of current
biomedical
research. The emergence
of state of the art culturing techniques is promoting the realization of the full potential
oí
pluripotent stem cells in basic and translational research and in cell-based therapies.
This comprehensive and authoritative atlas summarizes more than a decade of experience
accumulated by a leading research team in this field. Hands-on step-by-step guidance
for the derivation and culturing of human pluripotent stem ceils in defined conditions
(animal product-free, serum-free, feeder-free) and in non-adhesion suspension
culture are provided, as well as methods for examining pluripotency (embryoid body
and teratoma formation) and karyotype stability.
The Atlas of Human Pluripotent Stem Cells
·
Derivation and Culturing will serve as a
reference and guide to established researchers and those wishing to enter the promising
field of pluripotent stem cell research.
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| spelling | Atlas of human pluripotent stem cells derivation and culturing Michal Amit ; Joseph Itskovitz-Eldor [ed.] New York, NY [u.a.] Humana Press, Springer 2012 XIV, 142 S. zahlr. Ill. txt rdacontent n rdamedia nc rdacarrier Stem cell biology and regenerative medicine Labortechnik (DE-588)4123602-6 gnd rswk-swf Stammzelle (DE-588)4665329-6 gnd rswk-swf Stammzelle (DE-588)4665329-6 s Labortechnik (DE-588)4123602-6 s b DE-604 Amit, Michal Sonstige (DE-588)139597514 oth Itskovitz-Eldor, Joseph Sonstige (DE-588)1018432035 oth Erscheint auch als Online-Ausgabe 978-1-61779-548-0 Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024978372&sequence=000003&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024978372&sequence=000004&line_number=0002&func_code=DB_RECORDS&service_type=MEDIA Klappentext |
| spellingShingle | Atlas of human pluripotent stem cells derivation and culturing Labortechnik (DE-588)4123602-6 gnd Stammzelle (DE-588)4665329-6 gnd |
| subject_GND | (DE-588)4123602-6 (DE-588)4665329-6 |
| title | Atlas of human pluripotent stem cells derivation and culturing |
| title_auth | Atlas of human pluripotent stem cells derivation and culturing |
| title_exact_search | Atlas of human pluripotent stem cells derivation and culturing |
| title_full | Atlas of human pluripotent stem cells derivation and culturing Michal Amit ; Joseph Itskovitz-Eldor [ed.] |
| title_fullStr | Atlas of human pluripotent stem cells derivation and culturing Michal Amit ; Joseph Itskovitz-Eldor [ed.] |
| title_full_unstemmed | Atlas of human pluripotent stem cells derivation and culturing Michal Amit ; Joseph Itskovitz-Eldor [ed.] |
| title_short | Atlas of human pluripotent stem cells |
| title_sort | atlas of human pluripotent stem cells derivation and culturing |
| title_sub | derivation and culturing |
| topic | Labortechnik (DE-588)4123602-6 gnd Stammzelle (DE-588)4665329-6 gnd |
| topic_facet | Labortechnik Stammzelle |
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