Umbilical cord blood stem cells can expand hematopoietic and neuroglial progenitors in vitro.
The ability of hematopoietic tissue-derived adult stem cells to transdifferentiate into neural progenitor cells offers an interesting alternative to central nervous system (CNS)- or embryonic-derived stem cells as a viable source for cellular therapies applied to brain regeneration. Umbilical cord blood (CB) due to its primitive nature and it unproblematic collection appears as a promising candidate for multipotent stem cell harvest. We developed a negative immunomagnetic selection method that depletes CB from hematopoietic lineage marker-expressing cells, hence isolating a discrete lineage negative (LinNeg) stem cell population (0.1% of CB mononucleated cell [MCN] population). In liquid culture supplemented with thrombopoietin, flt-3 ligand, and c-kit ligand (TPOFLK), CB LinNeg stem cells could expand primitive nonadherent hematopoietic progenitors (up to 47-fold) and simultaneously produce slow-dividing adherent cells with neuroglial progenitor cell morphology over 8 weeks. Laser scanning confocal microscopy analysis identified these adherent cells to express glial fibrillary acidic protein (GFAP). Gene expression analysis showed upregulation of primitive neuroglial progenitor cell markers including, GFAP, nestin, musashi-1, and necdin. ELISA quantification of liquid culture supernatant revealed the in vitro release of transforming growth factor beta-1 (TGFbeta1), glial cell line-derived neurotrophic factor (GDNF) suggesting their contribution to CB LinNeg stem cell transdifferentiation into neuroglial progenitors. Our study supports that a single CB specimen can be pre-expanded in TPOFLK to produce both primitive hematopoietic and neuropoietic progenitors, hence widening CB clinical potential for cellular therapies.
McGuckin
Colin P
CP
King-George Laboratory, St. George's Hospital Medical School and Kingston University, London, UK. c.mcguckin@kingston.ac.uk
Forraz
Nicolas
N
Allouard
Quentin
Q
Pettengell
Ruth
R
D015415:Biomarkers; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005312:Fetal Blood / Q000166:cytology*; D015870:Gene Expression; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D007231:Infant, Newborn; D008565:Membrane Proteins / Q000494:pharmacology; D020932:Nerve Growth Factor / Q000032:analysis; D009414:Nerve Growth Factors / Q000032:analysis; D009457:Neuroglia / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D019089:Stem Cell Factor / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000032:analysis
D015415:Biomarkers; C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D008565:Membrane Proteins; D009414:Nerve Growth Factors; D019089:Stem Cell Factor; D016212:Transforming Growth Factor beta; C084881:flt3 ligand protein; D013926:Thrombopoietin; D020932:Nerve Growth Factor