Tchao, Jason
(2015)
Engineered Human Cardiac Tissue from Muscle Derived Stem Cells.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
Heart failure results in significant cardiomyocyte (CM) loss, and post-natal mammalian heart has limited regenerative capacity. Cellular cardiomyoplasty has emerged as a novel therapy to restore contractile function. A number of cell types illicit functional benefits through paracrine mechanisms, but cardiac stem cells are unique in their ability to preferentially differentiate down a cardiac lineage to replace lost CMs. However, cardiac stem cell isolation is highly invasive. Alternatively, skeletal myoblasts can be safely isolated and showed some benefits in clinical trials as donor muscle cells, but arrhythmias occurred due to lack of electric coupling with host cells. This limitation could be overcome by differentiating cells toward a cardiomyogenic lineage. Multipotent muscle derived stem cells (MDSC) are different from skeletal myoblasts and possess greater phenotypic plasticity. Our studies showed that cardiac and skeletal muscle share major genes/proteins during development in rodents, so it may be possible for human MDSCs to differentiate into CM-like cells under the appropriate conditions. My dissertation aims to develop approaches to differentiate human MDSCs into CM-like cells. Specifically, my work focuses on three aims: (I) to characterize the biochemical and functional properties of human MDSCs cultured in a 3-dimensional engineered muscle tissue (EMT) and examine whether it recapitulates properties of developing striated muscle; (II) to determine the potential for further CM differentiation under defined biophysical and chemical conditions; (III) to evaluate the potential of human MDSC derived cardiac progenitors to improve cardiac function in a human-rat xenograft model.
The results of my studies showed that human MDSCs in EMT beat spontaneously, displayed calcium transients, expressed cardiac-specific genes/proteins, and exhibited pharmacological responses similar to iPS cell-derived CMs. They also possessed characteristics of skeletal muscle including expression of MyoD, myogenin, and sk-fMHC. Their electrical coupling also remained immature. By temporally treating EMT with 4 chemical factors (4CF: miR-206 inhibitor, IWR-1, BMP4, and LiCl) and improving aggregation conditions, 4F-AEMT showed better muscle tissue formation and cardiac-like morphology with improved contractility, pharmacological responses, and electrical coupling. Although 4F-AEMT expressed MyoD and myogenin, it exhibited more cardiac-like function. Finally, human MDSC-aggregates showed evidence of survival and improved cardiac function in vivo.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
29 January 2015 |
| Date Type: |
Publication |
| Defense Date: |
24 March 2014 |
| Approval Date: |
29 January 2015 |
| Submission Date: |
5 June 2014 |
| Access Restriction: |
1 year -- Restrict access to University of Pittsburgh for a period of 1 year. |
| Number of Pages: |
141 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Swanson School of Engineering > Bioengineering |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
cardiac, tissue engineering, stem cells, MDSC, cardiomyocyte |
| Date Deposited: |
29 Jan 2016 06:00 |
| Last Modified: |
15 Nov 2016 14:20 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/21779 |
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