Skeletal Myogenic Differentiation of Mesenchymal Stem Cells Isolated from Human Umbilical Cord Blood

Abstract

Human umbilical cord blood (UCB) has been regarded as an alternative source for cell transplantation and cell therapy because of its hematopoietic and nonhematopoietic (mesenchymal) potential. Although there has been debate about whether mesenchymal stem cells (MSCs) are invariably present in UCB, several reports showed that MSC‐like cells could be consistently derived from human UCB and, moreover, could differentiate into various cells of a mesodermal origin. However, it remains unclear whether these UCB‐derived MSCs are also capable of differentiating into skeletal muscle cells. In this study, we isolated MSCs from human UCB and induced them to differentiate into skeletal muscle cells. During cell culture expansion, UCB‐derived mononuclear cells gave rise to adherent layers of fibroblast‐like cells expressing MSC‐related antigens such as SH2, SH3, α‐smooth muscle actin, CD13, CD29, and CD49e. More important, when these UCB‐derived MSCs were incubated in promyogenic conditions for up to 6 weeks, they expressed myogenic markers in accordance with myogenic differentiation pattern. Both flow cytometric and reverse transcriptase–polymerase reaction analyses showed that two early myogenic markers, MyoD and myogenin, were expressed after 3 days of incubation but not after 2 weeks. At week 6, more than half of UCB‐derived MSCs expressed myosin heavy chain, a late myogenic marker. Our results demonstrate that UCB‐derived MSCs possess a potential of skeletal myogenic differentiation and also imply that these cells could be a suitable source for skeletal muscle repair and a useful tool of muscle‐related tissue engineering.

Bone marrow‐derived very small embryonic‐like stem cells: Their developmental origin and biological significance

“…It is well known that CB-derived cells contribute to skeletal muscle (Gang et al, 2004;Brzoska et al, 2006), liver (Kakinuma et al, 2003;Newsome et al, 2003;Di Campli et al, 2004), neural tissue (Buzanska et al, 2002), and myocardium regeneration (Henning et al, 2004;Ma et al, 2005), and more importantly, recent multiorgan engraftment and differentiation has been achieved in goats after transplantation of human CB CD34 ϩ lin Ϫ cells (Zeng et al, 2006a). Furthermore, several groups of investigators described the presence of Oct-4 ϩ , Nanog ϩ , and SSEA-3/4 ϩ stem cells in human CB and umbilical cord matrix (Carlin et al, 2006).…”

Differential Gene Expression Profiling of Human Umbilical Cord Blood–Derived Mesenchymal Stem Cells by DNA Microarray

“…Six full-term UCB units, each containing about 80 mm of blood, were processed as previously described [24][25][26]. For the isolation of MSCs, MNCs were plated at a density of 1 × 10 6 cells per cm 2 and allowed to adhere to culture flasks for 5 days.…”

Limited plasticity of mesenchymal stem cells cocultured with adult cardiomyocytes

“…A vast literature reports about MSCs differentiation under defined in vitro conditions into the cells of several tissues, like osteocytes, chondrocytes, adipocytes [Pittenger et al, 1999], skeletal muscle fibers [Bhagavati and Xu, 2004;Gang et al, 2004], hepatocytes [Lee et al, 2004]. In particular, regarding heart repair, they have been shown to potentially originate both endothelial vascular cells and cardiomyocytes.…”