Kruglyakov Peter, Vijde S., Zinkova N., Polyntsev D., Kislyakova T.
(«Trans-Technologies», Ltd.; 189650, Russia, Saint-Petersburg, Pesochny, Leningradskaya str., 70/4 - 6.; e-mail: pitkus@alkorbio.ru)
Mesenchymal stem cells (MSC) are resident pluripotent cells of bone marrow stroma. MSC have the ability to differentiate into chondroblasts, adipocytes and osteoblasts. Their potential ability to differentiate neurons, glia and cardiomyocytes has been widely discussed recently.
MSCs were isolated from femoral bones of Wistar-Kyoto rats, expanded in culture and phenotyped by flow cytometry analysis of surface staining for negative (CD34 and CD45) and positive (CD90) markers. After the second passage cell culture consisted of 90-95% of CD90+ cells (mesenchymal stem cells) and 5-8% of CD45+ (haematopoietic cells). We demonstrated that CD90+/ CD45- cells were able to differentiate into chondrogenic, adipogenic and osteogenic lineages. During the process of differentiation into these lineages specific marker genes expression increased. In undifferentiated MSC fibronectin mRNA expression was on a high level. However, it decreased during differentiation into any of mentioned above lineage.
Cardiomyogenic differentiation was induced by incubation with 5'-azacitidine. Spontaneously contracting myotube-like structures were observed on the culture pellets and an increased expression level of 8 cardiomyocyte marker genes was shown. For the neuronal differentiation induction a growth factor neurotrophin-3 was applied. We showed neuronal differentiation of about 70% of the whole cell population. Differentiated cells demonstrated reaction with antibodies against neuron-specific nuclear protein Neuronal Nuclei (NeuN), neurofibrillary tangle protein (Tau) and myelin-olygodendrocyte specific protein (MOSP), but not with glial filament acidic protein (GFAP).
Thus, CD90+/ CD45- cells of rat bone marrow can be considered adult multipotent stem cells.
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