The demand for treatment strategies of musculoskeletal tissues is continuously growing, especially considering the increasing number of elderly people with degenerative diseases of the skeletal system. Despite major strides in the field of bone regenerative medicine during the years, current therapies, such as bone grafts, still have several limitations. Multipotent stem cells, such as mesenchymal stem cells (MSCs) are promising candidates for tissue repair because of their differentiation potential and their capacity to undergo extensive replication. However, isolating a homogeneous population of MSCs from multiple sources is an area that needs to be addressed. Also, the knowledge regarding the mechanisms and pathways that lead to the final osteogenic differentiation is still scarce. The following research is a feasibility study on a new isolation technique developed by our lab. The major focus of the research will be the isolation and characterization of mesenchymal stem cells from both adult bone marrow and umbilical cord blood using a novel isolation method based on immunodepletion. Furthermore we will look at the potential of these isolated MSCs to differentiate into mature, bone producing osteoblasts. The results of the studies showed that our novel isolation method allowed proliferation of a homogeneous MSC population. Our irnrnunodepleted MSCs were 99% double positive for antibodies CD44 and CD105 which are highly specific for multipotent MSCs while cells isolated using the plastic adherence method were only 43% double positive for the two MSC-specific markers. Homogeneous MSCs were derived from both adult bone marrow and umbilical cord blood using our isolation method. Utilizing the techniques of confocal microscopy, von Kossa staining, and RTPCR we also show that MSCs, upon stimulation with osteogenic supplements, differentiate into osteoblasts capable of being used for bone tissue engineering applications.

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