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N. Nourbakhsh, M.D.*, A. Talebi, M.D., B. Mousavi, M.D., F. Nadali, Ph.D.,
M. Torabinejad, M.D., Kh. Karbalaie, M.Sc., M.H. Nasr-Esfahani, Ph.D.*,
H.Baharvand, Ph.D.
* Corresponding Addresses:
P.O.Box: 19395-4644, Stem Cell Department, Royan Institute, ACECR, Tehran, Iran
Email: mh_nasr@med.mui.ac.ir
P.O.Box: 8174673461, Pediatric Department, Faculty of Medical Sciences, Isfahan
University of Medical Sciences, Isfahan, Iran
Email: nourbakhsh@dnt.mui.ac.ir
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Received: 4/Aug/2007, Accepted: 17/Feb/2008
Objective: The exfoliated human deciduous tooth (SHED) contain multipotent
stem cells that identified to be a population of highly proliferative and
clonogenic .These cells are capable of differentiating into a variety of cell
types including neural cells, adipocytes, and odontoblasts.
Material and Methods: Normal exfoliated human deciduous incisors
collected from six- to nine-years-old children. The pulp was separated from the
crown and digested with collagenase .Single cell solutions were cultivated in
α-MEM supplemented with ES-FCS. After two to three days, the cells reached
confluency and were trypsinized and cultured for further passages. The passage-4
cells were analyzed with CD34, CD45, CD105, CD166, CD31, CD90 and CD146 markers
that indicated these cells had a mesenchymal stem cell (MSC) identity. We
examined the cells for Alkaline Phosphatase activity to investigate the
mesenchymal (stromal) nature.Finally, the cells were differentiated into the
osteoblastic and adipocytic lineages in different subcultures and analysed by
RT-PCR and different staining protocols.
Results: Viable cells growing out of the explants showed elongated shapes
in
clusters. These cells showed alkaline phosphatase activity. Flow cytometry
results revealed high expression of pluripotent stem cell markers .In some area
of the osteoinductive cultures nodule-like structures were observed that showed
red mineralizing area upon staining with Alizarin Red.In adipogenic cultures
lipid vesicles appeared after five weeks of induction with Oil Red.
Conclusion: This study show that pulp contains cells with high plasticity
and proliferation capacity and can be easily isolated without any serious
intervention.
Keywords: Stem Cells, Differentiation, Tooth pulp, Flowcytometry
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