Journal List > J Korean Assoc Oral Maxillofac Surg > v.36(1) > 1032448

J Korean Assoc Oral Maxillofac Surg. 2010 Feb;36(1):7-15. Korean.
Published online February 28, 2010.  https://doi.org/10.5125/jkaoms.2010.36.1.7
Copyright © 2010 by The Korean Association of Oral and Maxillofacial Surgeons
A study on differentiation potency of adult stem cells from pulp, periodontal ligament, and dental follicle to osteoblast
Joong-Kyou Lee and Jae-Hoon Lee
Department of Oral and Maxillofacial Surgery, College of Dentistry, Dankook University, Korea.

Corresponding author: Jae-Hoon Lee. Department of Oral and Maxillofacial Surgery, College of Dentistry, Dankook University, 29 Anseodong, Choenan, Chungnam, 330-714, Korea. Tel: +82-41-550-1991~3, Fax: +82-41-551-8988, Email: lee201@dankook.ac.kr
Received December 18, 2009; Revised January 20, 2010; Accepted January 29, 2010.

Abstract

Complex human tissues harbor stem cells and precursor cells, which are responsible for tissue development or repair. Recently, dental tissues such as dental pulp, periodontal ligament (PDL), dental follicle have been identified as easily accessible sources of undifferentiated cells.

These tissues contain mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from pulp, PDL, and dental follicle and differentiate them into osteoblast and examine the bone induction capacity.

Dental pulp stem cell (DPSC), periodontal ligament stem cell (PDLSC), and dental follicle precursor cell (DFPC) were obtained from human 3rd molar and cultured. Each cell was analyzed for presence of stem cell by fluorescence activated cell sorter (FACs) against CD44, CD105 and CD34, CD45. Each stem cell was cultured, expanded and grown in an osteogenic culture medium to allow formation of a layer of extracellular bone matrix. Osteogenic pathway was checked by alizarin red staining, alkaline phosphatase (ALP) activity test and RT-PCR for ALP and osteocalcin (OCN) gene expression.

According to results from FACs, mesenchymal stem cell existed in pulp, PDL, and dental follicle. As culturing with bone differentiation medium, stem cells were differentiated to osteoblast like cell. Compare with stem cell from pulp, PDL and dental follicle-originated stem cell has more osteogenic effect and it was assumed that the character of donor cell was able to affect on differential potency of stem cell. From this article, we are able to verify the pulp, PDL, and dental follicle from extracted tooth, and these can be a source of osteoblast and stem cell for tissue engineering.

Keywords: Stem cells; Osteoblasts; Dental pulp; Periodontal ligament (PDL); Dental follicle

Figures


Fig. 1
Morphology of stem cells from dental tissues.(2-3 passages)

A, B: DPSC (× 200, × 100), C, D: PDLSC (× 200, × 100), E, F: DFPC (× 200, × 100).

(DPSC: dental pulp stem cell, PDLSC: periodontal ligament stem cell, DFPS: dental follicle precursor cell)

Click for larger image


Fig. 2
Characterization of DPSC, PDLSC, and DFPC immunophenotype in vitro.

(DPSC: dental pulp stem cell, PDLSC: periodontal ligament stem cell, DFPS: dental follicle precursor cell)

Click for larger image


Fig. 3
Morphology of cells in osteogenic culture media after 21 days (× 100).

A, C, E: DPSC, PDLSC, DFPC.

In universal culture media after 21 days (× 100).

B, D, F: DPSC, PDLSC, DFPC.

(DPSC: dental pulp stem cell, PDLSC: periodontal ligament stem cell, DFPS: dental follicle precursor cell)

Click for larger image


Fig. 4
Alizarin red staining of cultured cells in osteogenic media after 21 days.

A, C, E: DPSC, PDLSC, DFPC.

In universal culture media.

B, D, F: DPSC, PDLSC, DFPC.

(DPSC: dental pulp stem cell, PDLSC: periodontal ligament stem cell, DFPS: dental follicle precursor cell)

Click for larger image


Fig. 5
ALP activity test after 7, 14, 21 days of culture. A: Experimental group (osteogenic media), B: Control group (universal media).

(DPSC: dental pulp stem cell, PDLSC: periodontal ligament stem cell, DFPS: dental follicle precursor cell)

Click for larger image


Fig. 6
RT-PCR analysis for expression of osteoblast markers after 7, 14, 21 days of culture (exp: osteogenic media, control: universal media). A: ALP, B: OCN.

(ALP: alkaline phosphatase, OCN: osteocalcin, DPSC: dental pulp stem cell, PDLSC: periodontal ligament stem cell, DFPS: dental follicle precursor cell, GAPDH: glyceraldehyde 3-phosphate dehydrogenase)

Click for larger image

Tables


Table 1
Characteristics of samples. Sample number, sex and age, location of extracted teeth, and remark.
Click for larger image


Table 2
RT-PCR primer list and sequence.
Click for larger image


Table 3
ALP activity test (unit: concentration mM).
Click for larger image

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