Undifferentiated mouse mesenchymal stem cells spontaneously express neural and stem cell markers Oct-4 and Rex-1.
BACKGROUND
Previous adult stem cells studies have provided evidence that BM mesenchymal stem cells (MSC) exhibit multilineage differentiation capacity. These properties of MSC prompted us to explore the neural potential of MSC with a view to their use for the treatment of demyelinating disorders, such as multiple sclerosis. Indeed, issues such as the identification of a subset of stem cells that is neurally fated, methods of expansion and optimal stage of differentiation for transplantation remain poorly understood.


METHODS
In order to isolate mouse (m) MSC from BM, we used and compared the classic plastic-adhesion method and one depleting technique, the magnetic-activated cell sorting technique.


RESULTS
We established and optimized culture conditions so that mMSC could be expanded for more than 360 days and 50 passages. We also demonstrated that undifferentiated mMSC express the neural markers nestin, MAP2, A2B5, GFAP, MBP, CNPase, GalC, O1 under standard culture conditions before transplantation. The pluripotent stem cell marker Oct-4 and the embryonic stem cell marker Rex-1 are spontaneously expressed by untreated mMSC. The lineage-negative mMSC (CD5- CD11b- Ly-6G- Ter119- CD45R- c-kit/CD117-) overexpressed Oct-4, O1 and A2B5 in the first days of culture compared with the non-sorted MSC. Finally, we identified a distinct subpopulation of mMSC that is primed towards a neural fate, namely Sca-1+/nestin+ mMSC.


CONCLUSIONS
These results should facilitate the optimal timing of harvesting a neurally fated subpopulation of mMSC for transplantation into animal models of human brain diseases.
Lamoury
F M J
FM
Department of Neurology, Centre for Immunology, St Vincent's Hospital, Corner West and Boundary Streets, Sydney, NSW 2010, Australia. j.lamoury@cfi.unsw.edu.au
Croitoru-Lamoury
J
J
Brew
B J
BJ
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000032:analysis / Q000235:genetics; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics; D015415:Biomarkers / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation; D050989:GATA2 Transcription Factor / Q000235:genetics; D005698:Galactosylceramidase / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000235:genetics; D007621:Karyotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics; D064231:Nestin; D050814:Octamer Transcription Factor-3 / Q000032:analysis*; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D017479:Receptors, Platelet-Derived Growth Factor / Q000235:genetics; D019098:Telomerase / Q000378:metabolism; D014157:Transcription Factors / Q000032:analysis* / Q000235:genetics; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics
D018952:Antigens, CD34; D015415:Biomarkers; D050989:GATA2 Transcription Factor; C494712:Gata2 protein, mouse; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; C513672:Zfp42 protein, mouse; D014157:Transcription Factors; C497971:Flt3 protein, mouse; D017479:Receptors, Platelet-Derived Growth Factor; D051941:fms-Like Tyrosine Kinase 3; D019098:Telomerase; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D005698:Galactosylceramidase