Effects of Differentiated Placenta Stem Cell Conditioned Media on Proliferation and Migration of Human Dermal Fibroblasts
- 주제(키워드) differentiated stem cell , conditioned media , wound healing
- 발행기관 고려대학교 대학원
- 지도교수 김찬화
- 발행년도 2015
- 학위수여년월 2015. 8
- 학위구분 석사
- 학과 대학원 생명공학과
- 세부전공 의약생명공학전공
- 원문페이지 77 p
- 비고 비공개
- 실제URI http://www.dcollection.net/handler/korea/000000060207
- 본문언어 영어
- 제출원본 000045915075
초록/요약
Stem cells have a great skin regenerating potential, thus they are spotlighted in cosmetics and pharmaceutical field. Placenta stem cells (PSCs) are a type of mesenchymal stem cells which have a capacity of multipotency and secrete various factors that enhance wound healing process. Although placenta is a generally known as a discarded tissue, it has a good advantage due to its accesibility. Unlike existing method using undifferentiated stem cells, PSC were induced to differentiate keratinocyte. Expreriments were conducted whether secreted factors from differentiated placenta stem cells (DPSCs) could help wound healing process in human dermal fibroblasts and what composition of factors are made in conditioned media. To examine the effect of DPSCs conditioned media (CM) in wound healing process, proliferation and migration assay were conducted. Proliferation of fibroblasts was enhanced with 40 μg/ml DPSC-CM treatment. Migration rate of fibroblasts was also enhanced with 40 μg/ml DPSC-CM treatment. Moreover, in terms of mRNA expression level, DPSC-CM also promoted the production of extracelullar matrix (ECM) such as fibronectin, collagen and matrix metalloproteinases (MMPs). To better understand the effects of ECM related wound healing, the level of collagen-degradative enzyme (matrix metalloproteinase-1) were observed. Matrix metalloproteinase -1 expression was inhibited by DPSC-CM. Furthermore, using LC-MS/MS, secreted factors were identified and the conditioned media of PSCs and DPSCs were compared. In summary, the results of this study suggest that secreted factors of DPSCs contribute to the wound healing process of fibroblasts by promoting their synthesis of ECM proteins and enhancing their proliferative and migratory capacity. DPSC-CM offers good prospects to be utilized in the biopharmaceutical or cosmetic field due to its wound healing properties.
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Abstract
I. Introduction
II. Literature Review
2.1. Human Placenta
2.1.1. What is Human Placenta?
2.2. Human Placenta Stem Cell
2.2.1. What is Human Placenta Stem Cell?
2.3. Conditioned Medium
2.3.1. Conditioned medium
III. Materials and Methods
3.1. Concentration of conditioned medium
3.2. Cell culture
3.3. Cell proliferation assay
3.4. Cell migration assay
3.5. RT-PCR
3.6. In-gel digestions
3.7. LC-MS/MS
3.8. Statistical analysis.
IV. Results
4.1. Effects of CM on fibroblasts
4.2. ECM sysnthesis expression in Hs27 fibroblasts
4.3. Profiling of Placenta stem cell secretome
4.4. Comparative analysis of functional classification for secreted proteins
V. Discussion
VI. References
