Chapter title |
Human Embryonic and Hepatic Stem Cell Differentiation Visualized in Two and Three Dimensions Based on Serial Sections
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Chapter number | 128 |
Book title |
Human Embryonic Stem Cell Protocols
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Published in |
Methods in molecular biology, October 2014
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DOI | 10.1007/7651_2014_128 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2667-1, 978-1-4939-2668-8
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Authors |
Peter S. Vestentoft, Christian B. Brøchner, Niels Lynnerup, Claus Yding Andersen, Kjeld Møllgård, Vestentoft, Peter S., Brøchner, Christian B., Lynnerup, Niels, Andersen, Claus Yding, Møllgård, Kjeld |
Abstract |
Pluripotent human embryonic stem cells (hESCs) are characterized by two defining properties, self-renewal and differentiation. Self-renewing hESCs express transcription factors OCT4, SOX2, and NANOG, and surface markers SSEA-4 and TRA-1-60 and TRA-1-81 and their ability to differentiate into derivatives of the three germ layers show the differentiating potential. Studies suggest a certain microheterogeneity of the hESC colonies, in which not all cells in one colony of apparently undifferentiated cells express all the expected markers. We describe a technique to paraffin embed an entire hESC colony, and prepare 3-5 μm thick serial sections. Immunohistochemistry applied to individual sections produces a 2-dimensional survey of the developing hESC colony. Based on serial paraffin sections of the 2D-expression pattern, a new and useful 3D-visualization can be modeled. The actual 3D rendering of an entire colony is accomplished using 3D image processing software such as Mimics(®) or Amira(®). An extended version of this technique even allows for a high-magnification 3D-reconstruction of, e.g., hepatic stem cells in developing liver. These techniques combined allow for both a 2- and a 3-dimensional visualization of hESC colonies and stem cells in organs, which leads to new insights into and information about the interaction of stem cells with their surroundings. |
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Mendeley readers
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Student > Ph. D. Student | 2 | 25% |
Researcher | 2 | 25% |
Student > Postgraduate | 1 | 13% |
Student > Master | 1 | 13% |
Unknown | 2 | 25% |
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Medicine and Dentistry | 3 | 38% |
Computer Science | 2 | 25% |
Biochemistry, Genetics and Molecular Biology | 1 | 13% |
Engineering | 1 | 13% |
Unknown | 1 | 13% |