A simple and efficient method for generating Nurr1-positive neuronal stem cells from human wisdom teeth (tNSC) and the potential of tNSC for stroke therapy |
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| Authors: | Kuo-Liang Yang Mei-Fang Chen Chia-Hsin Liao Chen-Yoong Pang Py-Yu Lin |
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| Affiliation: | 1. Faculty of Medicine, University of Basel, Basel, Switzerland;2. Traslational Research laboratory, Albacete University Hospital, and Regional Biomedical Research Center, Castilla La Mancha University, Albacete, Spain;3. Department of Medical Oncology, Institute of Oncology Ljubljana, Ljubljana, Slovenia;4. Divisions of Medical Oncology and Hematology, Princess Margaret Cancer Centre, Department of Medicine, University of Toronto, Toronto, Canada;5. Department of Medical Oncology and Hematology, St. Claraspital, Basel, Switzerland;1. Department of Paediatrics I, Neonatology, University Hospital Essen, University Duisburg-Essen, Essen, Germany;2. Department of Neurology, University Hospital, Essen, Germany;3. Department of Hematology, West German Cancer Center, University Hospital of Essen, Essen, Germany;4. Institute of Medical Microbiology, University Hospital Essen, University Duisburg-Essen, Essen, Germany |
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| Abstract: | Background aimsWe have isolated human neuronal stem cells from exfoliated third molars (wisdom teeth) using a simple and efficient method. The cultured neuronal stem cells (designated tNSC) expressed embryonic and adult stem cell markers, markers for chemotatic factor and its corresponding ligand, as well as neuron proteins. The tNSC expressed genes of Nurr1, NF-M and nestin. They were used to treat middle cerebral artery occlusion (MCAO) surgery-inflicted Sprague–Dawley (SD) rats to assess their therapeutic potential for stroke therapy.MethodsFor each tNSC cell line, a normal human impacted wisdom tooth was collected from a donor with consent. The tooth was cleaned thoroughly with normal saline. The molar was vigorously shaken or vortexed for 30 min in a 50-mL conical tube with 15–20 mL normal saline. The mixture of dental pulp was collected by centrifugation and cultured in a 25-cm2 tissue culture flask with 4–5 mL Medium 199 supplemented with 5–10% fetal calf serum. The tNSC harvested from tissue culture, at a concentration of 1–2 × 105, were suspended in 3 µL saline solution and injected into the right dorsolateral striatum of experimental animals inflicted with MCAO.ResultsBehavioral measurements of the tNSC-treated SD rats showed a significant recovery from neurologic dysfunction after MCAO treatment. In contrast, a sham group of SD rats failed to recover from the surgery. Immunohistochemistry analysis of brain sections of the tNSC-treated SD rats showed survival of the transplanted cells.ConclusionsThese results suggest that adult neuronal stem cells may be procured from third molars, and tNSC thus cultivated have potential for treatment of stroke-inflicted rats. |
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