Minimally Manipulative Method for the Expansion of Human Bone Marrow Mesenchymal Stem Cells to Treat Osseous Defects

International Journal of Molecular Sciences

KristaL. Denning¹  LoganM. Lawrence¹  PierPaolo Claudio²  Andrew Cottrill³  Amrita Valluri³  Jagan Valluri³  Gaetano Marenzi⁴  JamesB. Day⁵ 

[1] Cabell Huntington Hospital Laboratory, Department of Pathology, Marshall University Joan C. Edwards School of Medicine, Huntington, WV 25705, USA;Department of BioMolecular Sciences, National Center for Natural Product Research, University of Mississippi, University, MS 39216, USA;Department of Biological Sciences, Marshall University, Huntington, WV 25705, USA;Department of Neuroscience, Reproductive, and Odontostomatologic Sciences, University of Naples “Federico II” 80131, Italy;Department of Orthopaedic Surgery, Marshall University Joan C. Edwards School of Medicine, Huntington, WV 25705, USA;

关键词: bone marrow mesenchymal stem cells;    autologous transplant;    regenerative medicine;    fracture healing;    bone defects;    traumatic injuries;   

DOI  :  10.3390/ijms20030612

来源: DOAJ

【 摘 要 】

Lack of standardization of clinically compliant culture protocols of mesenchymal stem cells for re-implantation in humans have hindered clinical progress in the field of tissue regeneration to repair maxillofacial and orthopedic defects. The goal of this study was to establish a clinically relevant osteogenic protocol for collection and expansion of autologous stem cells to be used at Marshall University for re-implantation and repair of maxillofacial and orthopedic conditions. Human bone marrow (hBM) samples were collected from patients undergoing intramedullary nail fixation for closed femoral fractures. hBM mesenchymal cells were expanded by growing them first in Petri dishes for two weeks, followed by a week of culture using Perfecta 3D Hanging Drop Plates^®. Various scaffold materials were tested and analyzed for cellular integration, vitality, and differentiation capacity of harvested hBM-MSCs including: 60/40 blend of hydroxyapatite biomatrix; Acellular bone composite discs; Allowash^®, cancellous bone cubes; PLGA (poly lactic-co-glycolic acid); and Woven chitin derived fiber. We found that the 3D spheroid culture allowed production of hBM mesenchymal cells that retained osteoblast differentiation capacity over a monolayer culture of hBM-MSCs without the need to use chemical or hormonal modulation. We also observed that hydroxyapatite and Allowash cancellous bone scaffolds allowed better cell integration and viability properties as compared to other materials tested in this study. In conclusion, the multimodal culture methodology we developed creates actively differentiating stem-cell spheroids that can then be readily utilized in clinical practices to improve the regeneration of tissues of the head and the body.

【 授权许可】

Unknown