공배양 및 물리적 환경하에서의 중간엽 줄기세포의 혈관 관련 세포로의 분화 경향 (The Effects of Co-Culture and Physical Stimulation on the Differentiation of Mesenchymal Stem Cell into Vascular Lineage Cells)

서지정보

· 발행기관 : 한국조직공학과 재생의학회
· 수록지 정보 : 조직공학과 재생의학 / 10권 / 1호 / 11 ~ 18페이지
· 저자명 : 박소희, 신지원, 김수향, 강윤경, 전강진, 김선연, 현진숙, 신정욱

초록

How to control the differentiation of mesenchymal stem cells (MSCs) into vascular lineage cells have still left much unclear even with numerous studies. Most in vitro studies on the differentiation of stem cells have utilized biochemical reagents. However, recent studies show the potentials of physical and co-culturing environments in control of differentiation. Therefore, we first factored out the in vivo conditions into biochemical reagents, co-culture,and physical stimulation. Then combinational effects of those factors on the differentiation of MSCs into vascular lineage cells were studied. For this EGM ®-2, co-culture, and shear stress were adopted to meet biochemical, neighboring cells, and physical condition, respectively. A total 4 experimental groups were set depending on co-culture and shear stress. Cell density was 1×104cells/cm2 at all group. For the co-cultures, MSCs were mixed with human umbilical vein endothelial cells (HUVECs) in the ratio of 1:2. The 12 dyne/cm2 of shear stress was engaged: 4 hr/day for 2 days followed by 48 hr after seeding. For the analyses, the immunofluorescent staining and PCR was performed to evaluate the differentiation of MSCs into vascular lineage cells. The co-culture group without shear stress showed the ECs-related markers even on the first day of the culture. And this was more observable on day 4. However,differentiation of MSCs into smooth muscle cells was found dominant when shear stress was engaged even MSCs were cultured with HUVECs and EGM ®-2. From this study we confirmed that physical stimulation (12 dyne/cm2 in our study) plays an important role in controlling differentiation of MSCs into vascular lineage cell types when MSCs were co-cultured with ECs.

영어초록

How to control the differentiation of mesenchymal stem cells (MSCs) into vascular lineage cells have still left much unclear even with numerous studies. Most in vitro studies on the differentiation of stem cells have utilized biochemical reagents. However, recent studies show the potentials of physical and co-culturing environments in control of differentiation. Therefore, we first factored out the in vivo conditions into biochemical reagents, co-culture,and physical stimulation. Then combinational effects of those factors on the differentiation of MSCs into vascular lineage cells were studied. For this EGM ®-2, co-culture, and shear stress were adopted to meet biochemical, neighboring cells, and physical condition, respectively. A total 4 experimental groups were set depending on co-culture and shear stress. Cell density was 1×104cells/cm2 at all group. For the co-cultures, MSCs were mixed with human umbilical vein endothelial cells (HUVECs) in the ratio of 1:2. The 12 dyne/cm2 of shear stress was engaged: 4 hr/day for 2 days followed by 48 hr after seeding. For the analyses, the immunofluorescent staining and PCR was performed to evaluate the differentiation of MSCs into vascular lineage cells. The co-culture group without shear stress showed the ECs-related markers even on the first day of the culture. And this was more observable on day 4. However,differentiation of MSCs into smooth muscle cells was found dominant when shear stress was engaged even MSCs were cultured with HUVECs and EGM ®-2. From this study we confirmed that physical stimulation (12 dyne/cm2 in our study) plays an important role in controlling differentiation of MSCs into vascular lineage cell types when MSCs were co-cultured with ECs.

참고자료

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