J Korean Assoc Maxillofac Plast Reconstr Surg.
2011 Sep;33(5):385-391.
Use of Peristeum as a Source of Endothelial-like Cells
- Affiliations
-
- 1Department of Oral and Maxillofacial Surgery, Gyeongsang National University School of Medicine and Institute of Health Sciences, Biomedical Center (BK21), Korea. surbyun@gsnu.ac.kr
- 2Department of Oral and Maxillofacial Surgery, School of Dentistry, Pusan National University, Korea.
- 3Clinical Research Institute, Gyeongsang National University Hospital, Korea.
- 4Department of Biochemistry, Gyeongsang National University School of Medicine and Institute of Health Sciences, Biomedical Center (BK21), Korea.
- 5Department of Oral and Maxillofacial Surgery, College of Medicine, Ulsan University, Korea.
- 6Maxillofacial Center, Onhospital, Korea.
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 37degrees 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 100microm 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.
MeSH Terms
-
Bone and Bones
Carbocyanines
Cell Culture Techniques
Coculture Techniques
Collagen
Collagenases
Drug Combinations
Durapatite
Endothelial Cells
Laminin
Lipoproteins
Magnetics
Magnets
Mandible
Mesenchymal Stromal Cells
Molar, Third
Nylons
Osteoblasts
Osteogenesis
Perchloric Acid
Periosteum
Proteoglycans
Carbocyanines
Collagen
Collagenases
Drug Combinations
Durapatite
Laminin
Lipoproteins
Nylons
Perchloric Acid
Proteoglycans
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