Comparative Analysis of Telomere Length, Telomerase and Reverse Transcriptase Activity in Human Dental Stem Cells

Abstract

Stem cells from dental tissues have been isolated and established for tooth regenerative applications. However, basic characterization on their biological properties still needs to be investigated before employing them for effective clinical trials. In this study, we compared the telomere length, relative telomerase activity (RTA), and relative reverse transcriptase activity (RRA) as well as the surface antigen profile and mesenchymal differentiation ability in human dental papilla stem cells (DPaSCs), dental pulp stem cells (DPuSCs), and dental follicle stem cells (DFSCs) with mesenchymal stem cells (MSCs) derived from bone marrow. Dental stem cells (DSCs) were strongly positive for cell surface markers, such as CD44 and CD90. However, slightly lower expression of CD105 was observed in DPaSCs and DPuSCs compared to DFSCs and MSCs. Following specific induction, DPaSCs, DFSCs, and MSCs were successfully differentiated into adipocytes and osteocytes. However, DPuSCS, in particular, were able to differentiate into adipocytes but failed to induce into osteogenic differentiation. Further, all DSCs, MSCs, and MRC-5 fibroblasts as control were investigated for telomere length by nonradioactive chemiluminescent assay, RTA by relative-quantitative telomerase repeat amplification protocol (RQ-TRAP), and RRA by PCR-based assay. Mean telomere lengths in DPaSCs, DPuSCs, DFSCs, and MSCs was ϳ11 kb, and the values did not differ significantly (p < 0.05) among the cells analyzed. RTA levels in DPaSCs were significantly (p < 0.05) higher than in MSCs, DPuSCs, DFSCs, and MRC-5 fibroblasts and among DSCs, DFSCs showed a significantly (p < 0.05) lower RTA. Moreover, RRA levels were significantly (p < 0.05) higher in DPaSCs, DPuSCs, and MSCs than in DFSCs. Based on these observations, we conclude that among DSCs, DPaSCs possessed ideal characteristics on telomere length, telomerase activity and reverse transcriptase (RTase) activity, and may serve as suitable alternative candidates for regenerative medicine.

Differentiation potential of mesenchymal stem cells isolated from human dental tissues into non-mesodermal lineage

“…In addition to MSCs isolated from the bone marrow, many groups have isolated MSCs from various tissues and organs of the body, which exists there, for regenerating damaged tissues. Till recently, human-derived MSCs were isolated from diverse tissues such as adipose, skin, umbilical cord matrix, dental pulp/papilla and others have fully demonstrated the cellular properties of MSCs for clinical and research applications (Jeon et al 2011a(Jeon et al , 2011bAkiyama et al 2012;Karaoz et al 2013). The various types of MSCs exhibited similar fibroblasts-like morphology and growth pattern following attachment onto plastic culture plates (Jeon et al 2011a(Jeon et al , 2011b.…”

“…Till recently, human-derived MSCs were isolated from diverse tissues such as adipose, skin, umbilical cord matrix, dental pulp/papilla and others have fully demonstrated the cellular properties of MSCs for clinical and research applications (Jeon et al 2011a(Jeon et al , 2011bAkiyama et al 2012;Karaoz et al 2013). The various types of MSCs exhibited similar fibroblasts-like morphology and growth pattern following attachment onto plastic culture plates (Jeon et al 2011a(Jeon et al , 2011b. The investigated MSCs also expressed the fundamental cellular properties of alkaline phosphatase activity, stem-cellspecific cell surface makers and pluripotent stem-cellspecific transcripts, such as SOX-2, OCT-4 and NANOG (Song et al 2011;Lee et al 2013;Patil et al 2014).…”

“…Further, it has been demonstrated that cells derived from dental tissues with the developing third molar (wisdom tooth) bud are a unique source of MSCs (Jeon et al 2011a(Jeon et al , 2011bPatil et al 2014). In our previous studies, MSCs derived from dental papilla tissue (DPSCs) of third molars showed outstanding cellular properties, telomerase activity, length of telomeric repeats, reverse transcriptase activity, cluster of differentiation (CD) surface markers and expression of transcripts, compared to those of bone marrow MSCs, adipose MSCs and dental pulp/ follicle MSCs (Jeon et al 2011a(Jeon et al , 2011b.…”

“…Even though maintenance of self-renewal capacity in various types of MSCs remains open to question, their cellular properties on differentiation capacity have been well demonstrated. Each of the MSCs derived from various types of tissues was induced into cell types of mesodermal lineage, such as bone, cartilage, fat, muscle, tendon, marrow stroma and others (Jeon et al 2011a(Jeon et al , 2011bSong et al 2011;Lee et al 2011). Due to the cellular features exhibited by MSCs, these cells have been successfully applied in many areas, such as clinical and research applications, including cell therapy in field of regenerative medicine, cell-based screening test for medication development, to study differentiation and gene regulation (Sensebe et al 2010;Wang et al 2011).…”

Extra-Telomeric Roles of Telomeric Proteins

“…Telomeres progressively shorten with each cell division in cultured primary human cells until a critically shortened length is reached, upon which the cells enter replicative senescence. Although the relevance of replicative senescence to aging in vivo remains poorly understood, numerous reports suggest that telomere shortening may be associated with organismal aging (Epel et al 2009;Monaghan 2010), with concomitant metabolic decline and increased risk for disease and death (Jaskelioff et al 2011;Jeon et al 2011). Likewise, shorter telomere length has been shown to be associated with age-related diseases including coronary heart disease (Calvert et al 2011;Krauss et al 2011), hypertension (Dimitroulis et al 2011), and dementia (Rolyan et al 2011;Takata et al 2011), as well as with a general risk for diseases such as insulin resistance and obesity Makino et al 2010;Njajou et al 2011).…”