2017 Cardiovascular Research Day Abstract Book
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Isolation and characterization of primary bone marrow mesenchymal stem cells.<br />
Eman El-sawalhy, MD 1 • Lakshman Chelvarajan, PhD 2 • Ahmed Abdel-Latif, MD, PhD 1<br />
1Division of <strong>Cardiovascular</strong> Medicine, Gill Heart Institute, University of Kentucky • 2 Saha<br />
cardiovascular research center, University of Kentucky<br />
Postdoc<br />
•Introduction: Myocardial infarction remains a major clinical problem and the leading cause of<br />
mortality in the world. Bone marrow derived stem cells have the capacity to participate in cardiac<br />
repair and regeneration of compromised heart muscle. The aim of this study is to generate highly<br />
enriched mesenchymal stem cells to inject 3x106 into the hearts of mice recovering from a<br />
myocardial infarction.<br />
•Methods: We isolated bone marrow (BM) from crushed long bones and hip bones of 6-8 weeks old<br />
GFP positive mice (C57BL/6). The stem cells were cultured in MesenCult media supplemented with<br />
MesenPure. Cells were cultured under either normoxic or hypoxic conditions (4% O2) for about<br />
two weeks. The cultures were then passaged when they reached 80% confluence. The cells were<br />
analyzed by flow cytometry using mesenchymal stem cell (CD90.2 and SCA-1) and hematopoietic<br />
(CD45) markers.<br />
•Results: As expected, the baseline bone marrow cells were negative for both stem cell markers,<br />
CD90 and Sca-1, while expressing the hematopoietic marker, CD45 (80%). During early expansion<br />
all the CD45– cells were Sca1+CD90–. Following subsequent passaging of the enriched cells, they<br />
began to express CD90. By the end of the second passage, the proportion of Sca1+CD90+<br />
mesenchymal stem cells was 65%, while cells expressing Sca-1 alone was down to 32%. Most of the<br />
live healthy cells remained GFP+ throughout the expansion and passaging.<br />
Conclusion: Mesenchymal stem cells can be isolated and enriched from murine bone marrow in<br />
sufficient amounts to be used in our in vivo study to enhance regeneration of cardiomyocytes and<br />
restore cardiac function after myocardial infarction.<br />
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