A Pilot Study of Primary Culture of Oral Epithelium using the Direct Explant Technique
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Authors
Massie, Jesse
Issue Date
2014-04-29
Type
Thesis
Language
en_US
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Abstract
Purpose – This investigation was a pilot study to determine the feasibility of using the direct explant technique for tissue that is typically discarded from extracted teeth as a source for mesenchymal stem cells (MSCs). The long-term goal of this research is to identify an easily obtainable source of MSC’s for new and innovative therapies for bone loss due to periodontal disease.|Methods – Gingiva was excised from extracted teeth obtained from Creighton University’s Oral Surgery Clinic. The tissue was cut into fragments and placed into 12 well collagen-coated plates. The tissue was allowed to set for a brief period of time and then 100ul of nutrient media was gently added to the wells. The plate was incubated at 37 °C with 5% CO2 in air for 24 hours. Following the 24 hour incubation, 1 ml of media was added to each well. The media was changed every three days. When significant numbers of cells had migrated from the explants, the cells were transferred to 25 cm flasks and expanded until adequate numbers were obtained for analysis|Results – This study demonstrates small amounts of attached gingiva are a suitable source for both epithelial and fibroblast/mesenchymal cells. Preliminary attempts to characterize the isolated cells suggest the presence of MSCs. Further studies to qualitatively and quantitatively characterize the resultant cell types are in progress. |Conclusion/Significance – The direct explant technique is a viable method for obtaining cells from small tissue samples obtained from teeth. If the presence of MSCs is confirmed, these cells can be manipulated to become osteoblasts, which are the cells that produce bone. The potential use of MSC’s as a biological therapy has clinical applications for both dental and medical science.
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Publisher
Creighton University
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Copyright is retained by the Author. A non-exclusive distribution right is granted to Creighton University and to ProQuest following the publishing model selected above.