세포외기질 단백질로 micropatterning 된 표면 위에서의 인간 성체 간엽 줄기세포의 형태변화와 세포 점착 차이에 관한 연구
- 발행기관 고려대학교 대학원
- 발행년도 2004
- 학위명 박사
- 학과 대학원:의학과 의용생체공학 전공
- 식별자(기타) DL:000014913801
- 서지제어번호 000045197258
초록/요약
Patterning of surface using soft lithography is useful tools for regulating cellular morphology as well as function. Microcontact printing technology was used for patterning extracellular matrix protein in this thesis. This pattern was used for controlling cellular adhesion as well as morphology of human mesenchymal stem cell. PDMS molds were designed for controlling cellular adhesion by varying stripe width from 5 to 50 ㎛ and fixing spacer to 20㎛. Glass surface was coated with extracellular matrix proteins such as fibronectin and vitronectin with different PDMS molds. Human mesenchymal stem cells were cultured on the patterned surfaces. Cells were aligned on the surfaces by changing cellular morphology. Actin filaments were aligned according to the protein stripe on the surfaces. The length of the actin fiber is longer as the stripe width is thinner. Proteins for focal contact such as vinculin, paxillin, and FAK were localized only on the ECM patterned surfaces. Especially, paxillin and FAK playing key role for focal adhesion were colocalized on the patterned surfaces. This study provides the controlling localization of focal adhesion proteins as well as morphology of human MSCs on the patterned surfaces. For further study, the effects of the localization of focal contacts proteins of MSCs to the stemness and differentiation will be investigated.
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목 차
Abstract 2
목 차 4
List of Table 6
List of Figure 7
Ⅰ. 서 론 10
1. 연구의 필요성 10
2. 이론적 배경 11
1) 줄기 세포 11
2) 세포외 기질 13
3) Microcontact Printing 15
Ⅱ. 연구 목적 17
Ⅲ. 실험 방법 18
1. Microcontact Printing 18
2. 인간 성체 간엽 줄기 세포의 분리 21
3. Fluorescence-Activated Cell Sorting (FACS) 분석 22
4. 패턴 위에서의 인간 성체 간엽 줄기세포 배양 23
5. 형광 면역 염색 23
6. 영상 분석 24
7. 광학 현미경 관찰 26
Ⅳ. 결과 27
1. 패턴 설계 27
2. Collagen 용액을 이용한 Microcontact Printing 결과 29
3. 인간 성체 간엽 줄기 세포의 확인 33
4. 단백질 패턴 위에서 배양한 세포의 형태 변화 35
5. 단백질 패턴 위에서 배양한 인간 성체 간엽 줄기세포 의 Focal Adhesion 41
6. 인간 성체 간엽 줄기세포의 Oct-4발현54
Ⅴ. 고 찰 56
Ⅵ. 결 론 60
참 고 문 헌 61
