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Use of Peristeum as a Source of Endothelial-like Cells  

Park, Bong-Wook (Department of Oral and Maxillofacial Surgery, Gyeongsang National University School of Medicine and Institute of Health Sciences, Biomedical Center (BK21))
Kim, Shin-Won (Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University)
Kim, Uk-Kyu (Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University)
Hah, Young-Sool (Clinical Research Institute, Gyeongsang National University Hospital)
Kim, Jin-Hyun (Clinical Research Institute, Gyeongsang National University Hospital)
Kim, Deok-Ryong (Department of Biochemistry, Gyeongsang National University School of Medicine and Institute of Health Sciences, Biomedical Center (BK21))
Sung, Iel-Young (Department of Oral and Maxillofacial Surgery, College of Medicine, Ulsan University)
Cho, Yeong-Cheol (Department of Oral and Maxillofacial Surgery, College of Medicine, Ulsan University)
Son, Jang-Ho (Department of Oral and Maxillofacial Surgery, College of Medicine, Ulsan University)
Kim, Jong-Ryoul (Maxillofacial Center, Onhospital)
Byun, June-Ho (Department of Oral and Maxillofacial Surgery, Gyeongsang National University School of Medicine and Institute of Health Sciences, Biomedical Center (BK21))
Publication Information
Maxillofacial Plastic and Reconstructive Surgery / v.33, no.5, 2011 , pp. 385-391 More about this Journal
Abstract
Purpose: The periosteum is a well-known source of osteogenic precursor cells for tissue-engineered bone formation. However, cultured endothelial or endothelial-like cells derived from periosteum have not yet been investigated. This study focused on endothelial-like cell culture from the periosteum. Methods: Periosteal tissues were harvested from the mandible during surgical extraction of lower impacted third molars. The tissues were treated with 0.075% type I collagenase in phosphate-buffered saline (PBS) for 1 hr at $37^{\circ}C$ to release cellular fractions. The collagenase was inactivated with an equal volume of DMEM/10% fetal bovine serum (FBS) and the infranatant was centrifuged for 10 min at 2,400 rpm. The cellular pellet was filtered through a $100{\mu}m$ nylon cell strainer, and the filtered cells were centrifuged for 10 min at 2,400 rpm. The resuspended cells were plated into T25 flasks and cultured in endothelial cell basal medium (EBM)-2. Results: Among the hematopoietic markers, CD146 was more highly expressed than CD31 and CD34. The periosteal-derived cells also expressed CD90 and CD166, mesenchymal stem cell markers. Considering that the expression of CD146 was constant and that the expression of CD90 was lower at passage 5, respectively, the CD146 positive cells in passage 5 were isolated using the magnetic cell sorting (MACS) system. These CD146 sorted, periosteal-derived cells formed tube-like structures on Matrigel. The uptake of acetylated, low-density lipoprotein, labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI-Ac-LDL) was also examined in these cells. Conclusion: These results suggest that the CD146-sorted positive cells can be referred to as periosteal-derived CD146 positive endothelial-like cells. In particular, when a co-culture system with endothelial and osteoblastic cells in a three-dimensional scaffold is used, the use of periosteum as a single cell source would be strongly beneficial for bone tissue engineering.
Keywords
Periosteum; Periosteal-derived endothelial like cells;
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