태아 연골유래 전구 세포의 각막 상피 세포로의 분화
Differentiation of fetal cartilage-derived progenitor cells (FCPCs) into corneal epithelial-like cells
- 주제(키워드) Fetal derived cell , Fetal cartilage-derived progenitor cell , cornea , corneal epithelial cell , differentiation
- 발행기관 아주대학교
- 지도교수 민병현
- 발행년도 2018
- 학위수여년월 2018. 2
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000027534
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
PURPOSE. Fetal cartilage-derived progenitor cells (FCPCs) emerged as a potential of viable cells for use in tissue engineering. But there have been no reports of differentiation into corneal epithelial cells using FCPCs. In this study, we have analyzed the differentiation capabilities of FCPCs towards differentiated corneal epithelial-like cells in order to establish the potential usefulness of these cells for corneal epithelium disease. METHODS. The FCPCs were isolated from the femoral head of the fetal cartilage tissue. The differentiation into corneal epithelial cells ability of FCPCs was evaluated using a differentiation-media at 2 days, 7 days post seeding. And, FCPCs-sheet, fabricated by FCPCs, was also used for differentiation assay. In vitro test, the results were evaluated by ICC and western blot for corneal epithelial cell markers (Pax6, ABCG2, p63, and CK3). For in vivo test, FCPCs-sheet was applied to treat the chemical burn rabbit model. There were 3 groups: 1) Normal, 2) Untreated group, and 3) FCPCs-sheet treated group (n = 6). At 1 week post transplantation, the healing ability was observed by histology (HE, IHC for CK3, human nucleic antibody). RESULTS. In vitro experiments, there were the morphological changes in FCPCs at 2 and 7 days of culture. The differentiated cells from FCPCs or FCPCs-sheet expressed the corneal epithelial cells markers. In vivo experiment, the adhesive ability and viability of FCPCs-sheet was showed. The healing ability of chemical burn model was also observed. CONCLUSIONS. This study found that FCPCs-sheet improves repair of corneal defect in chemical burn disease, probably by differentiation of FCPCs to corneal epithelium like cells.
more목차
1. Introduction 1
2. Materials and Methods 4
2.1 Isolation and culture of FCPCs 4
2.2 Differentiation of corneal epithelial-like cells 4
2.3 Fabrication of FCPCs-sheet 4
2.4 Immunocytochemistry 5
2.5 Flow Cytometry analysis 5
2.6 Western blot analysis 6
2.7 Histological and immunohistochemistry analysis 6
2.8 Animal model of chemical burned cornea epithelium 7
2.9 Statistical analysis 7
3. Results 9
3.1 Morphological features and changes in FCPCs 9
3.2 Immunocytochemistry analysis 11
3.3 Flow cytometry analysis 13
3.4 Western blot analysis 15
3.5 Fabrication of FCPCs-sheet 17
3.6 Transparency of FCPCs-sheet 19
3.7 Differentiation ability of FCPCs-sheet 21
3.8 Observation of gross images in vivo 23
3.9 Evaluation of FCPCs-sheet location and viability ability in vivo 25
3.10 Histological and immunohistochemistry analysis in vivo 27
4. Discussion 29
5. References 32
국문요약 38
