Transplantation of cryopreserved muscle cells in dilated cardiomyopathy: effects on left ventricular geometry and function.
OBJECTIVE
Cell transplantation to prevent congestive heart failure in patients with inherited dilated cardiomyopathy might require the use of noncardiac donor cells unaffected by the genetic defect and cryopreservation to permit cell storage until the time of transplantation. However, the effects of cryopreservation on peripheral muscle cells harvested from a cardiomyopathic recipient and their subsequent ability to restore cardiac structure and function after transplantation are unknown.


METHODS
Skeletal myoblasts and vascular smooth muscle cells from cardiomyopathic hamsters (delta-sarcoglycan-deficient BIO 53.58 hamster) and age-matched normal donor hamsters were isolated, expanded in culture, and cryopreserved. After reanimation in culture, cell morphology and growth rate were assessed and compared with values seen in noncryopreserved cells. A total of 4 x 10(6) previously cryopreserved skeletal myoblasts (n = 10) and vascular smooth muscle cells (n = 10) harvested from cardiomyopathic donors were then transplanted into the left ventricles of 17-week-old BIO 53.58 hamsters. Hearts injected with culture medium alone (n = 11) served as controls. Heart function was assessed 5 weeks after transplantation on a Langendorff apparatus, and left ventricular geometry was quantified by means of computerized planimetry. Staining with 5-bromo-2'-deoxyuridine identified the injected cells.


RESULTS
Vascular smooth muscle cells from cardiomyopathic donors had an abnormal morphology and diminished growth rates in culture compared with vascular smooth muscle cells from normal donors. These markers of injury were exacerbated by cryopreservation. In contrast, vascular smooth muscle cells from normal donors and skeletal myoblasts from either cardiomyopathic or normal donors appeared normal in culture and were unaffected by cryopreservation. Both cryopreserved vascular smooth muscle cells and skeletal myoblasts from cardiomyopathic donors formed a viable muscle-resembling tissue that prevented wall thinning, limited left ventricular dilatation, and preserved global systolic function in hamsters with a genetic dilated cardiomyopathy. However, attenuation of cardiac remodeling and preservation of global heart function was greater after skeletal myoblast transplantation compared with vascular smooth muscle cell transplantation in parallel to the in vitro morphologic and growth characteristics of these cells.


CONCLUSIONS
Cryostorage of healthy donor cells does not prevent the benefits of cell transplantation on limiting remodeling and preserving cardiac function in the failing heart. The health of donor cells in vitro predicts their subsequent benefits on cardiac structure and function after transplantation. Cryopreservation of donor cells might facilitate a clinically applicable and effective approach for ventricular restoration with cell-transplantation therapy for patients with inherited dilated cardiomyopathy.
Ohno
Nobuhisa
N
Division of Cardiac Surgery, Department of Surgery, Toronto General Hospital, University of Toronto, Ontario, Canada.
Fedak
Paul W M
PW
Weisel
Richard D
RD
Mickle
Donald A G
DA
Fujii
Takeshiro
T
Li
Ren-Ke
RK
D000704:Analysis of Variance; D000818:Animals; D002306:Cardiac Volume; D002311:Cardiomyopathy, Dilated / Q000601:surgery*; D002455:Cell Division; D017690:Cell Transplantation / Q000379:methods; D006224:Cricetinae; D015925:Cryopreservation / Q000379:methods*; D004195:Disease Models, Animal; D006334:Heart Function Tests; D007150:Immunohistochemistry; D008297:Male; D018482:Muscle, Skeletal / Q000166:cytology / Q000473:pathology / Q000637:transplantation; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000473:pathology* / Q000637:transplantation*; D010784:Photomicrography; D011336:Probability; D012016:Reference Values; D012680:Sensitivity and Specificity; D014182:Transplantation, Autologous; D016277:Ventricular Function, Left / Q000502:physiology*; D020257:Ventricular Remodeling / Q000502:physiology*