Isolation of Pulp Stem Cells and SEM Examination
Project no. 383 · Status: Kapanmış · 2014
Summary
The aim of this study was to describe in vitro isolation of dental pulp stem cells from human molar teeth and examination with Scanning electron microscopy. Dental pulps were obtained from sound human third molars extracted for therapeutic reasons. Extracted pulp tissue was minced into small pieces. The tissue blocks were cultured in the alpha modification of Eagle medium (Gibco-BRL, Grand Island, NY) supplemented with 20% fetal bovine serum and a complex of 100 U/mL penicillin G and 100 mg/mL streptomycin. Dental pulp stem cells (DPSCs) characterized by flow cytometric analysis of CD90, CD105, CD73, CD44, CD19, CD45 ve CD34, CD11b, CD14, CD19, CD79? ve HLA-DR. To control stem cell nature, cultured cells differentiated into osteocytes and adipocytes. Pulp derived stem cells were pooled to establish stem cell bank. After 14 days, samples were then examined using a scanning electron microscope at various magnifications. Cells showed good proliferation and attachment to culture flasks. According to flow cytometric analysis results, CD105, CD73, CD90 and CD44 were positive while others were negative. Majority of the cells displayed fibroblast-like morphology at 7 th day of culture. Cells showed good proliferation rate, and particularly after 14 days of culture period. After 21 days, these primary cells reached 95% confluence and showed epithelioid and polygonal shape and formed colonies. The nature of dentin surface itself presented good adhesive characteristics for stem cells. Because root canal dentin is a target tissue of regenerative endodontic therapy, developing approaches to increase the adhesion and proliferation capacity of stem cells into the dentin lead to better clinical interventions