Culture of skeletal myoblasts from human donors aged over 40 years: dynamics of cell growth and expression of differentiation markersReportar como inadecuado




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Journal of Translational Medicine

, 3:21

First Online: 12 May 2005Received: 09 February 2005Accepted: 12 May 2005DOI: 10.1186-1479-5876-3-21

Cite this article as: Baj, A., Bettaccini, A.A., Casalone, R. et al. J Transl Med 2005 3: 21. doi:10.1186-1479-5876-3-21

Abstract

BackgroundLocal myogenesis, neoangiogenesis and homing of progenitor cells from the bone marrow appear to contribute to repair of the infarcted myocardium. Implantation into heart tissues of autologous skeletal myoblasts has been associated with improved contractile function in animal models and in humans with acute myocardial ischemia. Since heart infarction is most prevalent in individuals of over 40 years of age, we tested whether culture methods available in our laboratory were adequate to obtain sufficient numbers of differentiated skeletal myoblasts from muscle biopsy specimens obtained from patients aged 41 to 91.

Methods and resultsNo matter of donor age, differentiated skeletal muscle cells could be produced in vitro in amounts adequate for cellular therapy ≥300 millions. Using desmin as a cytoplasmic marker, about 50% cultured cells were differentiated along myogenic lineages and expressed proteins proper of skeletal muscle myosin type I and II, actin, actinin, spectrin and dystrophin. Cytogenetic alterations were not detected in cultured muscle cells that had undergone at least 10 population doublings. Molecular methods employed for the screening of persistent viral infections evidenced that HCV failed to replicate in muscle cells cultured from one patient with chronic HCV infection.

ConclusionThe proposed culture methods appear to hold promise for aged patients not only in the field of cardiovascular medicine, but also in the urologic and orthopedic fields.

KeywordsCellular therapy Stem cells Heart failure Karyotype Human viruses List of abbreviations usedBFGFbasic fibroblast growth factor

DMdifferentitation medium

DMEM-F12Dulbecco-s modified Eagle-s medium plus Ham-s F12 medium

EGFepidermal growth factor

FBSfetal bovine serum

FITCfluoresceine isothiocyanate

HCVhepatitis C virus

IFAimmunofluorescence assay

PMproliferation medium

QFQquinacrine chromosome banding

SCsatellite cells

Electronic supplementary materialThe online version of this article doi:10.1186-1479-5876-3-21 contains supplementary material, which is available to authorized users.





Autor: Andreina Baj - Alessia A Bettaccini - Rosario Casalone - Andrea Sala - Paolo Cherubino - Antonio Q Toniolo

Fuente: https://link.springer.com/







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