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Human Adipose Tissue Derived Stem Cells Differentiate into Epidermal Cell and Fibroblasts

Author: HanNa
Tutor: LiuXiaoMing
School: Dalian Medical University
Course: Dermatology and Venereology
Keywords: Adipose mesenchymal stem cells Differentiation Epidermal cells Fibroblasts
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 9
Quote: 0
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Abstract


Background:linical injury to the skin, such as chronic ulcers, burns, trauma requires immediate coverage to facilitate repair and restore skin function. Recently,Regenerative medicine that uses the the body’s own stem cells and growth factors provides a new opinion for repair of damaged tissue, and provides a new approach for cell-based therapy.Many studies have shown that stem cell therapy is clinically applied as a safe and effective method for repair of several types of tissue damage.Objective: ADSCs are differentiated in induction medium of growth factors containing EGF, bFGF, IGF - 1 ,TGF-β1etc in vitro, The isolation and proliferation of ADSCs , and the expression of surface molecule of ADSCs are detected,.And the level of transcription and protein are detected after induction.Methods:we adopt regenerative medicine which combines adult stem cells and growth factors,which means the combination of human adipose mesenchymal stem cells and EGF, bFGF, IGF - 1, TGFβ1. we isolated ADSCs, detected the cell surface marker such as CD34、CD90、CD106 in P3、P10,and analyzed its growth kinetics in P3、P5、P10。We have found no markedly difference in growth kinetics and cell surface marker between P 3 and P10. ADSCs was differentiated in induction medium of growth factors containing EGF, bFGF, IGF - 1 etc in vitro, the expression of CK 19 was detected , and cells were observed through transmission electron microscope(TEM). TGFβ1 and bFGF were used to induce ADSCs into fibroblasts,and analysizing the transcription and protein level of collagen type I.And cells were observed through TEM.Results:(1)ADSCs have the strongly positive expression of CD90,weakly positive expression of CD34 and negative expression of CD106. We have found no markedly difference in growth kinetics and cell surface marker between P 3 and P10.(2)ADSCs was differentiated in induction medium of growth factors containing EGF, bFGF, IGF - 1 etc in vitro. After 3 days, some cells were from fusiformis into irregular, after 5 days, ADSCs presented growth characteristics more resembling a monolayer culture of epithelial cells. Immunofluorescence results showed that 15.6% of ADSCs have the expression of cytokeratin19. After 15 days,we can see melanosome、keratohyaline granule through TEM(.3) TGFβ1 and bFGF were used to induce ADSCs into fibroblasts . After 7 days, RT-PCR identified collagen typeⅠmRNA expression more than twice than control group in differentiated cells. Immunofluorescence results showed that ADSCs had the higher expression of collagen type I than control group. After 15 days,we can see collagen microfibrils through TEM.Conclusions: This study,we adopt regenerative medicine which combines adult stem cells and growth factors,which means the combination of human adipose mesenchymal stem cells and EGF, bFGF, IGF - 1, TGFβ1,and experimental results show that ADSCs could differentiate into epidermal cells and fibroblasts in certain conditions. Thus, this study not only provides the sources of cells for the skin’s regeneration, but also provides theoretical basis for the clinical application of repairing skin’s damage.

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