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a Department of Anatomy and Physiology and
b Department of Animal Science, Kansas State University, Manhattan, Kansas, USA;
c Department of Anatomy and Histology, Faculty of Veterinary Medicine, Kafr-El-Sheikh, Tanta University, Tanta, Egypt
Key Words. Stem cell plasticity • Telomerase • c-kit • Myofibroblast • Neural induction
Kathy E. Mitchell, Ph.D., 228 Coles Hall, Department of Anatomy and Physiology, Kansas State University, Manhattan, Kansas 66506-5802, USA. Telephone: 785-532-4825; Fax: 785-532-4557; e-mail: mitchell{at}vet.ksu.edu
We have identified an easily attainable source of primitive, potentially multipotent stem cells from Wharton’s jelly, the matrix of umbilical cord. Wharton’s jelly cells have been propagated in culture for more than 80 population doublings. Several markers for stem cells, including c-kit (CD117), and telomerase activity are expressed in these cells. Treatment with basic fibroblast growth factor overnight and low-serum media plus butylated hydroxyanisole and dimethylsulfoxide induced Wharton’s jelly cells to express a neural phenotype. Within several hours of this treatment, Wharton’s jelly cells developed rounded cell bodies with multiple neurite-like extensions, similar to the morphology of neural stem cells. Neuron-specific enolase (NSE), a neural stem cell marker, was expressed in these cells, as shown by immunocytochemistry. Immunoblot analysis showed similar levels of NSE expression in both untreated and induced Wharton’s jelly cells. After 3 days, the induced Wharton’s jelly cells resembled bipolar or multipolar neurons, with processes that formed networks reminiscent of primary cultures of neurons. The neuron-like cells in these cultures stained positively for several neuronal proteins, including neuron-specific class III ß-tubulin, neurofilament M, an axonal growth-cone-associated protein, and tyrosine hydroxylase. Immunoblot analysis showed increasing levels of protein markers for mature neurons over time postinduction. Markers for oligodendrocytes and astrocytes were also detected in Wharton’s jelly cells. These exciting findings show that cells from the matrix of umbilical cord have properties of stem cells and may, thus, be a rich source of primitive cells. This study shows their capacity to differentiate into a neural phenotype in vitro.
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