혈관내피유사세포 채취의 원천으로 골막의 활용
Use of Peristeum as a Source of Endothelial-Like Cells
박봉욱, 김신원, 김욱규, 하영술, 김진현, 김덕룡, 성일용, 조영철, 손장호, 김종렬, 변준호,
소속 상세정보
박봉욱 ( Park Bong-Wook ) - 경상대학교 의학전문대학원 구강악안면외과학교실
김신원 ( Kim Shin-Won ) - 부산대학교 치의학전문대학원 구강악안면외과학교실
김욱규 ( Kim Uk-Kyu ) - 부산대학교 치의학전문대학원 구강악안면외과학교실
하영술 ( Hah Young-Sool ) - 경상대학교병원 임상의학연구소
김진현 ( Kim Jin-Hyun ) - 경상대학교병원 임상의학연구소
김덕룡 ( Kim Deok-Ryong ) - 경상대학교 의학전문대학원 생화학교실
성일용 ( Sung Iel-Yong ) - 울산대학교 의과대학 울산대학교병원 구강악안면외과학교실
조영철 ( Cho Yeong-Cheol ) - 울산대학교 의과대학 구강악안면외과학교실
손장호 ( Son Jang-Ho ) - 울산대학교 의과대학 구강악안면외과학교실
김종렬 ( Kim Jong-Ryoul ) - 온종합병원 턱얼굴수술센터
변준호 ( Byun June-Ho ) - 경상대학교 의학전문대학원 구강악안면외과학교실
KMID : 0360120110330050385
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.
키워드
Periosteum; Periosteal-derived endothelial Like cells
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