Stem cells from human extracted deciduous teeth (SHED) have the ability to multiply much faster and double their population in culture at a greater rate, indicating that it may be in a more immature state than other type of adult stem cells. Mesenchymal stem cells (MSC) from human primary molars were isolated and cultured in media supplemented with 20% fetal bovine serum. The MSCs were confirmed using CD 105 and CD 166 and the identification of the osteoblast cells were done using reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Differentiated osteoblast cells (DOC) were characterized by alkaline phosphotase and von Kossa staining followed by immunocytochemistry staining using osteocalcin and osteonectin antibodies. Further validation of SHED was done by RT-PCR to detect the presence of insulin-like growth factor 2 (IGF-2) and discoidin domain tyrosine kinase-2 (DDTK-2) transcripts, while the presence of Runx-2 mRNA was used to characterize DOC. The results showed that SHED was found positive for CD 105 and CD 166 and could differentiate into osteoblast, bone forming cells. The findings revealed the presence of distinct MSC population which had the capability to generate living human cells that could be a possible source for tissue engineering in the future.
The aim of this study was to isolate stem cells from dental pulp of primary molars and incisors to be used as possible source for tissue engineering. Human primary molars and incisors were collected from subjects aged 4-7 year-old under standardized procedures. Within 24 hours, the tooth was cut at the cemento-enamel junction using hard tissue material cutter. The dental pulp tissue was extracted, digested and then cultured in Alpha Modified Eagles's Medium (α-MEM) supplemented with 20% FCS, 100 mM L-ascorbic acid 2-phosphate, 200 mM L-glutamine and 5000 units/ml Penicillin/Streptomycin. The cells were observed daily under the microscope until confluence. Children's tooth pulp- derived progenitor cells were found positive for stem cell markers CD105 and CD166, which are consistent with the finding for mesenchymal stem cells (MSCs) from bone marrow.