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Min-Ju Jhin1, 2, Young-Sung Kim1, 3
, Su-Hwan Kim1, Kyoung-Hwa Kim1, Chul-Woo Lee1, Ki-Tae Koo1, Tae-Il Kim1, Yang-Jo Seol1, Young Ku1, In-Chul Rhyu1, Chong-Pyoung Chung1, Yong-Moo Lee1
, Su-Hwan Kim1, Kyoung-Hwa Kim1, Chul-Woo Lee1, Ki-Tae Koo1, Tae-Il Kim1, Yang-Jo Seol1, Young Ku1, In-Chul Rhyu1, Chong-Pyoung Chung1, Yong-Moo Lee1
Abstract
Purpose
The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow.
Methods
Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation.
Results
PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite/β-tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation.
Figures and Tables
Figure 1
The cultured canine PDLSCs form adherent clonogenic cell clusters of fibroblast-like cells (A) Colony forming Unit (Original magnification ×40) (B) typical fibroblast-like cells (Original magnification ×100).
Figure 2
Osteogenic differentiation (A) PDLSC (B) ABSC (C) BMSC (Alizarin red S, Original magnification ×100).
Figure 3
Adipogenic differentiation (A) PDLSC (B) ABSC (C) BMSC (Oil Red O staining, Original magnification ×100).
Figure 4
Chodrogenic differentiation (A) PDLSC (B) ABSC (C) BMSC (Alcian Blue staining, Original magnification ×200).
Figure 5
Flow cytometric analysis of PDLSC(A) 12.96% of PDLSC were positive staining for CD146 (B) 99.76% for CD44 (C) 99.40% for CD90 (D) 0.25% for STRO-1 (E) 5.85% for CD45 (F) 5.57% for CD34.
Figure 6
Flow cytometric analysis of ABSC (A) 38.97% of ABSC were positive staining for CD146 (B) 99.38% for CD44 (C) 99.24% for CD90 (D) 0.23% for STRO-1 (E) 9.56% for CD45 (F) 7.20% for CD34.
Figure 7
Flow cytometric analysis of BMSC (A) 11.49% of BMSC were positive staining for CD146 (B) 99.50% for CD44 (C) 86.39% for CD90 (D) 3.02% for STRO-1 (E) 3.47% for CD45 (F) 2.88% for CD34.
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