INSTANT STEM CELL THERAPY FOR ORTHOPEDIC APPLICATIONS: Automated separation of mesenchymal cells from bone marrow aspirates

INTRODUCTION: Mesenchymal stem cells (MSC) constitute an essential element in many applications in regenerative medicine. In orthopedics, they are used as osteoinductive component for bone regeneration. One source for MSC is bone marrow (BM) aspirate which contains concentrations of 10-100 MSC per Mio BM cells. This low number requires an efficient isolation procedure and usually an in-vitro expansion to increase the total number of MSC before use. Routinely, the mononuclear cell (MNC) fraction that contains MSCs is isolated by centrifugation in a Ficoll gradient and subsequently plated on tissue culture plastic to separate adherent and non adherent cell. All the procedures have to be carried out by skilled personnel under sterile conditions considering GMP regulations if such cell are to be used routinely in clinical settings. The aim of our study was to adapt and test a fully automated and closed separation system allowing for separating BM into MNC fractions in the operating room within 1 hour. These fractions were biochemically and biologically characterized and compared to MSC separated by manual Ficoll procedures and to native BM cells.