Identification of stemness and differentially expressed genes in human cementum-derived cells

EunHye Lee; Young-Sung Kim; Yong-Moo Lee; Won-Kyung Kim; Young-Kyoo Lee; Su-Hwan Kim

doi:10.5051/jpis.2102600130

Identification of stemness and differentially expressed genes in human cementum-derived cells

J Periodontal Implant Sci. 2021 Oct;51(5):329-341. doi: 10.5051/jpis.2102600130.

Authors

EunHye Lee¹, Young-Sung Kim², Yong-Moo Lee³, Won-Kyung Kim², Young-Kyoo Lee⁴, Su-Hwan Kim⁵

Affiliations

¹ Dental Research Institute, Seoul National University, Seoul, Korea.
² Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.
³ Department of Periodontology and Dental Research Institute, Seoul National University School of Dentistry, Seoul, Korea.
⁴ Department of Dentistry, Uijeongbu Eulji Medical Center, Eulji University, Uijeongbu, Korea.
⁵ Department of Periodontics, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea. shkimperio@amc.seoul.kr.

Abstract

Purpose: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored.

Methods: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed.

Results: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs.

Conclusions: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.

Keywords: Dental cementum; MicroRNAs; Microarray analysis; RNA, messenger; Stem cells.

Grants and funding

2013-541/Asan Institute for Life Sciences, Asan Medical Center/Korea