기계적 자극이 3차원 공배양을 통한 중간엽 줄기세포의 chondrogenesis에 미치는 영향 (Effects of Mechanical Stimulation on Chondrogenesis of Mesenchymal Stem Cells via 3D Co-culturing System)

서지정보

· 발행기관 : 한국조직공학과 재생의학회
· 수록지 정보 : 조직공학과 재생의학 / 6권 / 4호 / 739 ~ 745페이지
· 저자명 : 김동화, 신지원, 박소희, 신정욱, 허수진

초록

The goal of this study is to investigate the effects of mechanical stimulation on chondrogenesis of mesenchymal stem cells(MSCs) via 3D co-culturing system. The chondrocytes and MSCs were isolated and cultured from New Zealand White male rabbits. Cells were maintained in chondrogenic differentiation media containing 10 ng/mL TGF-β1. MSCs and chondrocytes were suspended in different alginate beads respectively. Those two types of beads were separated by a track-etched membrane of 3 μm pore. The intermittent hydrostatic pressure(2 min pressurizing/15 min resting, 0.2 MPa) was applied for 3 days, starting on the third day after seeding and ending on the sixth day for 4 hrs/day. Proliferation and production of glycosaminoglycan(GAG) were evaluated along with
immunocytochemistry observations. The rate of MSC proliferation tended to decrease during the culture period, while that of chondrocytes increased regardless of mechanical stimulation. However, on 7 day the mechanical stimulation affected proliferation of chondrocytes. The differentiation of MSCs and chondrocytes were affected by the
mechanical stimulation on 14 day. When amount of GAG in each group was normalized by its DNA quantity to investigate differentiation activity of each cell, the efficacy of mechanical stimulation was observed after 7 day in MSCs. These results suggested that during co-culture with stimulation some secretion from chondrocytes, which are fully developed, tended to suppress the proliferation of MSCs while tended to promote differentiation of MSCs. In addition, the efficacy of mechanical stimulation was confirmed through this study.

영어초록

The goal of this study is to investigate the effects of mechanical stimulation on chondrogenesis of mesenchymal stem cells(MSCs) via 3D co-culturing system. The chondrocytes and MSCs were isolated and cultured from New Zealand White male rabbits. Cells were maintained in chondrogenic differentiation media containing 10 ng/mL TGF-β1. MSCs and chondrocytes were suspended in different alginate beads respectively. Those two types of beads were separated by a track-etched membrane of 3 μm pore. The intermittent hydrostatic pressure(2 min pressurizing/15 min resting, 0.2 MPa) was applied for 3 days, starting on the third day after seeding and ending on the sixth day for 4 hrs/day. Proliferation and production of glycosaminoglycan(GAG) were evaluated along with
immunocytochemistry observations. The rate of MSC proliferation tended to decrease during the culture period, while that of chondrocytes increased regardless of mechanical stimulation. However, on 7 day the mechanical stimulation affected proliferation of chondrocytes. The differentiation of MSCs and chondrocytes were affected by the
mechanical stimulation on 14 day. When amount of GAG in each group was normalized by its DNA quantity to investigate differentiation activity of each cell, the efficacy of mechanical stimulation was observed after 7 day in MSCs. These results suggested that during co-culture with stimulation some secretion from chondrocytes, which are fully developed, tended to suppress the proliferation of MSCs while tended to promote differentiation of MSCs. In addition, the efficacy of mechanical stimulation was confirmed through this study.

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