Isolation of oligodendrocyte-like cells from human umbilical cord blood.
BACKGROUND
As human umbilical cord blood (UCB) is known to be a rich source of progenitor cells, the prospect of isolating a subset of these cells that could differentiate into cells of non-hematopoietic lineages suggests a therapeutic use for patients with inherited lysosomal and peroxisomal storage diseases currently treated with UCB transplantation.


METHODS
Oligodendrocyte-like cells were isolated from UCB by density-gradient centrifugation and expanded using selective media. We then characterized this population of cells using standard immunohistochemical staining methods for neural cell proteins and polymerase chain reaction (PCR) to detect RNA sequences for myelin basic protein (MBP). We also developed a functional assay demonstrating myelination of neurons in vitro.


RESULTS
Cells with oligodendrocyte-like morphology were reproducibly cultured ex vivo from fresh human UCB. Cells stained positively for multiple oligodendria cell markers (O1, MBP and CNPase) via immunohistochemical staining and flow cytometry. PCR confirmed the presence of MBP and CNPase mRNA. A further in vitro functional assay demonstrated the myelination of mature neuronal cells from the brain of a myelin-deficient murine model co-cultured with the oligodendrocyte-like cells.


CONCLUSIONS
After human UCB transplant, donor-derived cells have been noted to migrate to the brain over time. Although is not known whether these cells solely deliver enzyme replacement or a subset engrafts and differentiates into mature neural cells, the clinical improvements noted in these patients suggest a potential role for targeted cellular therapy. Oligodendrocyte-like cells isolated ex vivo and expanded from human UCB could provide a potential cellular therapy for patients with demyelinating or dismyelinating diseases.
Tracy
E
E
Pediatric Blood and Marrow Transplant Program, Duke University Medical Center, Durham, North Carolina 27710, USA.
Aldrink
J
J
Panosian
J
J
Beam
D
D
Thacker
J
J
Reese
M
M
Kurtzberg
J
J
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000943:Antigens, Differentiation; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000276:immunology; D019070:Cell Lineage / Q000276:immunology; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D009185:Myelin Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D009836:Oligodendroglia / Q000166:cytology* / Q000378:metabolism; D011247:Pregnancy
D000943:Antigens, Differentiation; D009185:Myelin Proteins; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases