title: Human Embryonic and Hepatic Stem Cell Differentiation Visualized in Two and Three Dimensions Based on Serial Sections.
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.
authors.affiliation: Dako Denmark A/S, Produktionsvej 42, 2600, Glostrup, Denmark.
authors.forename: Peter S
authors.initials: PS
authors.lastname: Vestentoft
authors.forename: Christian B
authors.initials: CB
authors.lastname: Brøchner
authors.forename: Niels
authors.initials: N
authors.lastname: Lynnerup
authors.forename: Claus Yding
authors.initials: CY
authors.lastname: Andersen
authors.forename: Kjeld
authors.initials: K
authors.lastname: Møllgård
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D000066449:Human Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008099:Liver / Q000166:cytology*; D016612:Paraffin Embedding