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Title:	Neurogenic, chondrogenic and osteogenic analyses of mouse dental pulp stem cells
Authors:	Shabnam Kermani (P48707)
Supervisor:	Shahrul Hisham Zainal Ariffin, Prof Dr
Keywords:	Dental Pulp Stem cells Research Mesenchymal Stem Cells
Issue Date:	13-Sep-2012
Description:	Dental pulp stem cells (DPSC) do not require general anesthesia during extraction as compared to bone marrow stem cells. The aim of this study was to isolate mouse dental pulp stem cells and to differentiate them to neurons, chondrocytes, osteoblasts and osteoclasts. In the present study, the differentiation capability of DPSC in mice aged 6 to 8 weeks was investigated. The research focused on the expression of markers using reverse transcriptase polymerase chain reaction (RT-PCR) in mouse stem cells as well as the expression intensity of markers for nerve cells, i.e. Nestin and Tub 3 in the absence of the neural growth factor. To induce chondrocyte differentiation, DPSC were cultured in chondrogenic medium (Zen-Bio, Inc) for 21 days. The expression of cell-surface antigen markers for DPSC; Cd146 and Cd166 as well as chondrocyte markers; Coll І and Coll ІІ markers was examined by RT-PCR. To induce osteoblast differentiation, 50 μg/mL ascorbic acid and 10 mM β-glycerophosphate were added to basal medium whereas for osteoclast differentiation, 10 ng/mL RANKL and 5 ng/mL M-CSF were added to the basal medium. OPN and CatK markers were used to investigate osteoblast and osteoclast differentiation respectively. To study cell survival during differentiation to nerve cells, chondrocytes and osteoblasts, MTT method was employed. Alkaline phosphatase (ALP) activity was enzymologically assayed for chondrocyte and osteoblast differentiations and Tartrate-Resistant Acid Phosphatase (TRAP) assay was used for osteoclast differentiation. Expression of microtubule associated protein 2 (MAP2) was determined using MAP2 antibodies by immunocytochemistry. In RT-PCR study, DPSC expressed Cd146 and Cd166 markers but cells did not express Cd38 blood cell markers indicating that these cells belonged to mesenchymal cells. In differentiation to neural cells, Nestin markers were observed before and after differentiation, but the expression marker after differentiation was significantly increased, whereas Tub 3 marker was only observed after differentiation. Immunocytochemical staining revealed that neural cells were capable of producing MAP2 on the fifth day of neural induction in the absence of growth factors. Cell viability analyses, in control group (cells cultured in proliferation medium) showed that these cells maintained their growth rate. Coll І marker was expressed after 14 days of chondrogenic induction whereas Coll ІІ being mature chondrocyte cell markers were expressed only after 21 days treatment in chondrocyte induction medium. OPN marker was observed after 21 days whereas CatK marker was not detected after osteoclast differentiation. Cell viability assay showed that differentiated cells treated with chondrogenic medium and osteogenic differentiation medium maintained their growth rate in comparison with control group. However, after 14 days, growth rate decreased but was still viable. This showed that the cells survived during chondrocyte and osteoblast differentiation. ALP assay result showed that, after 10 days of culturing DPSCs with chondrogenic induced medium most of the cells became alkaline phosphatase positive compared with control group cells whereas in osteoblast medium the cells became alkaline phosphatase positive after 21days as compared with cells in the control group. In conclusion, this study shows that extracted dental pulp contains stem cells which can be successfully differentiated into nerve cells, chondrocytes and osteoblasts.,Tesis ini tiada perakuan deklarasi pelajar,Doktor Falsafah
Pages:	209
Call Number:	QH588.S83K446 2012 tesis
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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