Determination of the specific phenotype markers of ex vivo expanded human peridontal ligament cells

2005 Fiscal Year Final Research Report Summary

• Project/Area Number: 16592078
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Periodontal dentistry
• Research Institution: Showa University
• Principal Investigator: KOBAYASHI Makoto Showa University, Periodontology, Lecturer, 歯学部, 講師 (80186767)
• Co-Investigator(Kenkyū-buntansha): MURAKAMI Shinya Osaka University, Periodontology, Professor, 歯学部, 教授 (70239490) ; MIYAZAWA Yasushi Showa University, Pefiodontology, Lecturer, 歯学部, 講師 (90219775) ; OHAZAMA Atsushi Showa University, Periodontology, Assistant, 歯学部, 助手 (40266169) ; OKAMATSU Yoshimasa Showa University, Periodontology, Assistant, 歯学部, 助手 (50286845) ; TAKIGUCHI Takashi Showa University, Periodontology, Assistant, 歯学部, 助手 (60317576)
• Project Period (FY): 2004 – 2005
• Keywords: Periodontal Regeneration / Human Periodontal Ligament Cells / Mesenchymal Stem Cells / Cell Surface Antigens / Multilineage Differentiation

Research Abstract

The transplantation of cultured human periodontal ligament (HPL) cells with scaffolds into periodontal defects appears to be a powerful strategy to promote periodontal regeneration. To clarify the specific phenotype markers of the cultured HPL cells, we investigated here the frequency of HPL cells expressed marker antigens of various cell types and the frequency of three types of mesenchymal progenitor cells existed within HPL cells, compared with human mesenchymal stem cells (HMSCs).
Frequency of HPL cells expressed markers of various cell types
In HPL cells, Ratio of cells expressed MSC markers (CD105,CD44,CD29) or integrins (CD49c, CD49e) were high, and those of cells expressed a hematopoietic stem cell marker (CD117), hemotopoietic cell markers (CD34,CD45) or a neuronal stem cell marker (AC133) were low levels.
Frequency of three types of mesenchymal progenitor cells existed within cultured HPL cells
The expression profile of MSC markers ; CD45 negative (CD45^-),CD90 positive (CD90^+), CD105^+,CD 146^+ or STRO-1^+, in HPL cells was similar to that in HMSCs, suggesting that many constituent cells of HPL cell cultures possess MSC phenotype. However, the potential of multilineage differentiation in HPL cells was different from that in HMSCs. HMSCs highly differentiated into three mesenchymal cell types, osteoblasts, adopocytes and chondrocytes under each lineage-specific culture condition. Also in HPL cells, the increase of ALP^+ cell number, mineralized nodule formation and expression of bone-specific genes was detected. Whereas, the number of adpocytes induced in HPL cells cultures was a few, and the development of cartilage containing type II collagen were not detected in HPL cell pellet cultures. These results suggest that the cultured HPL cells contain many osteoblast lineage cells and small number of adipocyte precursors, while chondrocyte precursors might exist at slight levels or not within the cells. Further, ALP^- cell fraction isolated from HPL cells contained relatively more adipocyte precursors.