Efficient in vitro generation of adult multipotent cells from mobilized peripheral blood CD133+ cells.
OBJECTIVE
To generate non-haematopoietic tissues from mobilized haematopoietic CD133(+) stem cells.


METHODS
Mobilized peripheral blood CD133(+) cells from adult healthy donors were used. In vitro ability of highly enriched CD133(+) cells from mobilized peripheral blood to generate multipotent cells, and their potential to give rise to cells with characteristics of neuroectoderm, endoderm and mesoderm layers was investigated.


RESULTS
We found that a recently identified population of CD45(+) adherent cells generated in vitro after culture of highly purified CD133(+) cells for 3-5 weeks with Flt3/Flk2 ligand and interleukin-6 can, in presence of the appropriate microenvironmental cues, differentiate into neural progenitor-like cells (NPLCs), hepatocyte-like cells and skeletal muscle-like cells. We have termed them to be adult multipotent haematopoietic cells (AMHCs). AMHC-derived NPLCs expressed morphological, phenotypic and molecular markers associated with primary neural progenitor cells. They can differentiate into astrocyte-like cells, neuronal-like cells and oligodendrocyte-like cells. Moreover, AMHC-derived NPLCs produced 3,4-dihydrophenylalanine and dopamine and expressed voltage-activated ion channels, suggesting their functional maturation. In addition, AMHC-derived hepatocyte-like cells and skeletal muscle-like cells, showed typical morphological features and expressed primary tissue-associated proteins.


CONCLUSIONS
Our data demonstrate that AMHCs may therefore serve as a novel source of adult multipotent cells for autologous replacement cell therapies.
Kuçi
S
S
University Children's Hospital, Department of Hematology/Oncology, University of Tübingen, Tübingen, Germany. selim.kuci@kgu.de
Kuçi
Z
Z
Schmid
S
S
Seitz
G
G
Müller
I
I
Dufke
A
A
Leimig
T
T
Murti
G
G
Jurecic
R
R
Schumm
M
M
Lang
P
P
Bruchelt
G
G
Bader
P
P
Klingebiel
T
T
Niethammer
D
D
Handgretinger
R
R
D000071916:AC133 Antigen; D000328:Adult; D015703:Antigens, CD / Q000276:immunology*; D001483:Base Sequence; D002454:Cell Differentiation; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D004295:Dihydroxyphenylalanine / Q000096:biosynthesis; D004298:Dopamine / Q000096:biosynthesis; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000276:immunology; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000071916:AC133 Antigen; D015703:Antigens, CD; D017931:DNA Primers; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D004295:Dihydroxyphenylalanine; D004298:Dopamine