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골수 간엽줄기세포로부터 분화된 연골세포, 골세포, 혈관내피세포를 이용한 골조직 및 연골조직의 재생 원문보기

  • 저자

    송창애

  • 학위수여기관

    慶北大學校 大學院

  • 학위구분

    국내석사

  • 학과

    의학과

  • 지도교수

  • 발행년도

    2004

  • 총페이지

    35 p.

  • 키워드

    골조직 연골조직 골수 간엽줄기세포;

  • 언어

    kor

  • 원문 URL

    http://www.riss.kr/link?id=T10062285&outLink=K  

  • 초록

    Most current strategies for tissue engineering have been changed to use adult stem cells as a cell source. Mesenchymal stem cells(MSCs) from bone marrow are the progenitors of multiple lineages and seem to be the one of the best candidates to regenerate injured tissue. However, recent advances in application of MSCs toward large tissue regeneration are faced with lack of vascularity. The aim of this study was to identify the role of co-seeding differentiated mesenchymal cells and endothelial cells from MSCs onto scaffold in constructing tissue-engineered bone and cartilage for promoting osteogenesis, chondrogenesis, and angiogenesis. MSCs were generated out of mononuclear bone marrow cells from healthy donors separated by density gradient centrifugation. They were able to differentiate into osteogenic cells, chondrogenic cells, endothelial cells after cultivation in respective media. Osteogenic cells, chondrogenic cells were co-cultured with endothelial cells from MSCs for evaluation of biologic character in vitro and preparing seeding in vivo. Cell-scaffold complexes formed by seeding individually cultured cells and co-cultured cells on PLGA(polylatic-co-glycolic acid) and fibrin glue were implanted into the back of nude mice. After 4 weeks implantation, cell-scaffold complexes were taken out and analyzed histologically. The gene expressions of cells used were analyzed by RT-PCR using osteopontin, aggrecan, CD133, KDR/Flk1,CD31. In vitro differentiation, RT-PCR analysis not only showed the well differentiated expression patterns on each marker generally, but also CD133, KDR/Flk1 marker gene expressions of co-cultured differentiated osteogenic/endothelial and chondrogenic/endothelial cells were more increased. In vivo study, it was demonstrated positive patterns of each organ-specific marker(collagen type I and II) immunohistochemically. The scaffold complexes with co-cultured differentiated osteogenic/endothelial and chondrogenic/endothelial cells showed more capillary formation in histological findings. Endothelial marker gene expressions of co-cultured cell-PLGA complexes were more increased than co-cultured cell-fibrin glue complexes. Osteopontin, aggrecan gene expressions of co-cultured cell-scaffold complexes were more increased than individually cultured cell-scaffold complexes in RT-PCR analysis. When differentiated mesenchymal cells were co-cultured with endothelial cells, the expressions of each specific marker genes were more increased, that is, the formation of each tissues was more predominant. In conclusion, these results suggest that endothelial cell from human mesenchymal stem cells has a very important role in tissue generation. Furthermore, it is expected that differentiated endothelial cells from MSCs may be the one of essential component for tissue-engineered construct using MSCs.


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