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J Thorac Cardiovasc Surg 2005;129:904-911
© 2005 The American Association for Thoracic Surgery

Cardiothoracic Transplantation

Quantitative analysis of survival of transplanted smooth muscle cells with real-time polymerase chain reaction

Division of Cardiovascular Surgery and the Toronto General Research Institute, Toronto General Hospital, and Division of Cardiac Surgery, the University of Toronto, Toronto, Ontario, Canada

Received for publication September 10, 2003; revisions received June 9, 2004; accepted for publication June 22, 2004.

^_* Address for reprints: Ren-Ke Li, MD, PhD, Toronto General Hospital, NU 1-115A, 200 Elizabeth St, Toronto, Ontario, M5G 2C4, Canada (E-mail: Renkeli{at}uhnres.utoronto.ca).

BACKGROUND: Cell transplantation improves heart function after myocardial infarction. This study investigated the survival of implanted cells in normal and infarcted myocardium.

METHODS: Male rat aortic smooth muscle cells were cultured. For the in vitro study, male smooth muscle cells mixed with female smooth muscle cells or male smooth muscle cells injected into a piece of female rat myocardium were used to evaluate the accuracy of quantitative real-time polymerase chain reaction to measure Y chromosomes. For the in vivo study, 2 million live or dead male smooth muscle cells were injected into normal or infarcted female myocardium. At 1 hour and 1 and 4 weeks after transplantation, hearts, lungs, and kidneys were harvested for measurement of Y chromosomes.

RESULTS: In vitro, the accuracy of polymerase chain reaction measurement was excellent in cultured cells (r² = 0.996) and the myocardium (r² = 0.786). In vivo, 1 hour after 2 x 10⁶ cell implantation, live cell numbers decreased to 1.0 ± 0.2 x 10⁶ and 1.1 ± 0.3 x 10⁶, and dead cell numbers decreased to 0.9 ± 0.2 x 10⁶ and 0.8 ± 0.2 x 10⁶ in the normal and infarcted myocardium, respectively (P < .01 for all groups). Lungs and kidneys contained 8.5% and 1.5% of the implanted cells, but no cells were detected at 1 week. At 1 week, no dead smooth muscle cells were detected in the normal or infarcted myocardium. The numbers of live cells at 1 and 4 weeks were 0.48 ± 0.06 x 10⁶ and 0.27 ± 0.07 x 10⁶ in normal myocardium and 0.29 ± 0.08 x 10⁶ and 0.18 ± 0.05 x 10⁶ in infarcted myocardium.

CONCLUSIONS: One hour after implantation, only 50% of smooth muscle cells remained in the implanted area. Some implanted cells deposited in other tissue. Implanted cell survival progressively decreased during the 4-week study.

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