Cytokine preconditioning promotes codifferentiation of human fetal liver CD133+ stem cells into angiomyogenic tissue.
BACKGROUND
CD133 (AC133) is a surface antigen that defines a broad population of stem cells, including myogenic and endothelial progenitors. CD133+ cells are rare in adult tissues, and the factors that support their differentiation into mature angiomyogenic cells are not known. These hurdles have hampered the use of CD133+ cells for therapeutic purposes. Because human fetal liver is a rich source of CD133+ cells, we sought to identify the growth factors that promote codifferentiation of these cells into angiogenic and myogenic cells.


RESULTS
Human fetal liver CD133+ and CD133- cell subpopulations were cultured with 5'-azacytidine or vascular endothelial growth factor (VEGF165) and/or brain-derived nerve growth factor (BDNF). CD133+ but not CD133- cells from human fetal liver codifferentiated into spindle-shaped cells, as well as flat adherent multinucleated cells capable of spontaneous contractions in culture. The resulting spindle-shaped cells were confirmed to be endothelial cells by immunohistochemistry analysis for von Willebrand factor and by acetylated LDL uptake. Multinucleated cells were characterized as striated muscles by electron microscopy and immunohistochemistry analysis for myosin heavy chain. Presence of VEGF165 and BDNF significantly enhanced angiomyogenesis in vitro. Inoculation of cells derived from CD133+ cells, but not CD133- cells, into the ear pinna of NOD/SCID mice resulted in the formation of cardiomyocytes, as identified by immunostaining with cardiac troponin-T antibody. These cells generated electrical action potentials, detectable by ECG tracing.


CONCLUSIONS
CD133 defines a population of human fetal liver cells capable of differentiating into both angiogenic and myogenic cells. Preconditioning of these CD133+ cells with VEGF165 and BDNF enhances the angiomyogenesis. CD133+ fetal liver cells ultimately may be used for therapeutic angiomyogenesis.
Shmelkov
Sergey V
SV
Weill Medical College of Cornell University, Division of Hematology-Oncology, New York, NY 10021, USA.
Meeus
Sarah
S
Moussazadeh
Nelson
N
Kermani
Pouneh
P
Rashbaum
William K
WK
Rabbany
Sina Y
SY
Hanson
Marilee A
MA
Lane
William J
WJ
St Clair
Ryan
R
Walsh
Kathryn A
KA
Dias
Sergio
S
Jacobson
Jason T
JT
Hempstead
Barbara L
BL
Edelberg
Jay M
JM
Rafii
Shahin
S
D000071916:AC133 Antigen; D000200:Action Potentials; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D001374:Azacitidine / Q000494:pharmacology; D015415:Biomarkers; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects; D004431:Ear, External; D042783:Endothelial Cells / Q000737:chemistry / Q000166:cytology*; D004730:Endothelium, Vascular / Q000166:cytology*; D020869:Gene Expression Profiling; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000196:embryology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009124:Muscle Proteins / Q000096:biosynthesis / Q000235:genetics; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology; D010455:Peptides / Q000032:analysis; D011971:Receptors, Immunologic / Q000378:metabolism; D051116:Receptors, Scavenger; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*; D014183:Transplantation, Heterologous; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*; D014841:von Willebrand Factor / Q000032:analysis
D000071916:AC133 Antigen; D015703:Antigens, CD; D015415:Biomarkers; D019208:Brain-Derived Neurotrophic Factor; D006023:Glycoproteins; D008077:Lipoproteins, LDL; D009124:Muscle Proteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D011971:Receptors, Immunologic; D051116:Receptors, Scavenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C032629:acetyl-LDL; D014841:von Willebrand Factor; D001374:Azacitidine