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Rapid and Specific Immunomagnetic Isolation of Mouse Primary Oligodendrocytes
Published on: May 21, 2018
Isolation of oligodendrocyte-like cells from human umbilical cord blood
E Tracy1, J Aldrink, J Panosian
1Pediatric Blood and Marrow Transplant Program, Duke University Medical Center, Durham, North Carolina 27710, USA.
Insights
Human umbilical cord blood yields oligodendrocyte-like cells that can myelinate neurons in vitro. These cells show therapeutic potential for treating demyelinating diseases.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Regenerative Medicine
Background:
- Human umbilical cord blood (UCB) is a rich source of progenitor cells.
- Potential exists for non-hematopoietic differentiation of UCB cells.
- UCB transplantation treats inherited lysosomal and peroxisomal storage diseases.
Purpose of the Study:
- To isolate and characterize oligodendrocyte-like cells from human UCB.
- To assess the therapeutic potential of these cells for neural repair.
Main Methods:
- Isolation of oligodendrocyte-like cells from UCB via density-gradient centrifugation and selective media expansion.
- Characterization using immunohistochemistry for neural markers and RT-PCR for myelin basic protein (MBP) and CNPase.
- In vitro functional assay for neuronal myelination.
Main Results:
- Reproducible ex vivo culture of oligodendrocyte-like cells from human UCB.
- Positive staining for oligodendrocyte markers (O1, MBP, CNPase) and confirmed mRNA expression.
- Demonstrated myelination of neurons from a myelin-deficient murine model in vitro.
Conclusions:
- UCB-derived oligodendrocyte-like cells can be expanded ex vivo.
- These cells exhibit functional myelination capabilities in vitro.
- Potential cellular therapy for demyelinating diseases.
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.
Discussion:
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.
