Paraffin-embedded manipulated blastocysts: a tool to demonstrate stem cell plasticity?
One of the big question marks in current stem cell research is whether there is true plasticity of adult progenitor cells (APC) or if cell fusion is the principle source of the supposed plasticity. The generation of chimeras by injecting adult progenitor cells into blastocysts is not new. This paper describes an efficient embedding technique for murine blastocysts injected with human APC. This method could help in establishing a novel tool to analyse the process of plasticity, if it truly exists. If this is the case, this technology could be of great help to characterize surface markers of stem cells in great detail. On the other hand, fusion of cells could also be investigated. A system of embedding blastocysts was set up using paraffin for further analysis by means of light microscopy and immunohistochemistry. The embedding of the chimaeras consists of fixing them first with paraformaldehyde in phosphate-buffered saline (PFA/PBS), embedding them in gelatine, fixing the gelatine block with PFA/PBS and finally fixing the gelatine block in a Petri dish by embedding it in paraffin. Using this protocol, the morphology of the blastocysts is well preserved.
Zech
N H
NH
Department of Obstetrics and Gynaecology, University Hospital, Zurich, Switzerland. Nicolas.Zech@usz.ch
Koestenbauer
S
S
Vanderzwalmen
P
P
Schoonjans
L
L
Danloy
S
S
Zech
H
H
Blaschitz
A
A
Dohr
G
G
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002678:Chimera; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D016612:Paraffin Embedding / Q000379:methods*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D033581:Stem Cell Transplantation