Preferential enrichment and characterization of human dental pulp stem cell via a genetic engineering-based approach

2016 Fiscal Year Final Research Report

• Project/Area Number: 25463192
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthodontics/Pediatric dentistry
• Research Institution: Kagoshima University
• Principal Investigator: Inada Emi 鹿児島大学, 医歯学域附属病院, 助教 (30448568)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: 歯髄幹細胞 / PiggyBacシステム / 選択的濃縮 / アルカリフォスファターゼ

Outline of Final Research Achievements

Alkaline phosphatase (ALP) is a useful marker for stem cells that differentiate into human deciduous tooth dental pulp cells (HDDPCs). In this study, we attempted to enrich for ALP-positive cells from HDDPCs via a genetic engineering-based approach. We employed a PiggyBac (PB)-based gene transfer system, which is known to be effective for the enrichment of transfectants. We found that this system is useful for acquiring transfectants when HDDPCs are transfected with PB vectors carrying cDNA for enhanced green fluorescent protein (EGFP). Subsequently, a PB vector carrying the EGFP cDNA and a puromycin resistance gene under the control of the ALP promoter was introduced into HDDPCs together with a transposase expression vector. Unfortunately, this caused accidental arrest of cell proliferation. To overcome this problem, we successfully created a HDDPC-derived immortalized cell line, which will be used as a source for ALP-positive HDDPCs.

Free Research Field

小児歯科