A rapid method for obtaining mesenchymal stem cells and platelets from bone marrow aspirate

Abstract

Mesenchymal stem cells (MSCs) and platelet‐rich plasma (PRP) are currently used alone or in combination for therapeutic applications especially for bone repair. We tested whether MSCs can be isolated from bone marrow (BM) aspirate using a commercially available kit commonly used to obtain PRP from peripheral blood (PB). Results revealed that mononuclear cells and platelets from both PB and BM could be efficiently isolated by obtaining a mononuclear and platelet rich fraction (PB‐MPRF and BM‐MPRF, respectively). Starting with comparable volumes, the number of platelets increased 1.5‐fold in BM‐MPRF compared to PB‐MPRF. The number of clonogenic cells in BM‐MPRF samples was significantly higher than whole BM samples as revealed by CFU‐F assay (54.92 ± 8.55 CFU‐F/1.5 x 105 nucleated cells and 32.50 ± 12.43 CFU‐F/1.5 x 105 nucleated cells, respectively). Cells isolated from BM‐MPRF after in vitro expansion fulfilled the definition of MSCs by phenotypic criteria, and differentiated along osteogenic, adipogenic and chondrogenic lineages following induction. Results showed that the kit isolated MSCs and platelets from BM aspirate. Isolated MSCs were further expanded in a laboratory and BM‐MPRF was used clinically following BM withdrawal for rapid intra‐operative cell therapy for the treatment of bone defects. Copyright © 2012 John Wiley & Sons, Ltd.

Protein kinase B/AKT isoform 2 drives migration of human mesenchymal stem cells

“…MSC resulted positive for CD44, CD73, CD90, CD105, CD146 and negative for CD34 and CD45 (17). The multilineage potential was evaluated by differentiation assays as previously described (18). After the appropriate stimulations, cells positively differentiated in osteogenic, adipogenic and condrogenic lineages.…”

Needle Biopsy Adequacy in the Era of Precision Medicine and Value-Based Health Care

“…Cells for Culture, Xenografts, Cell Therapy, or Single-Cell Analysis.-Needle biopsy to retrieve marrow stem cells is a long-established application. 51 Needle biopsies are increasingly used to retrieve tumor cells for xenograft diagnostic studies. [52][53][54][55][56] Ongoing adequacy issues include retrieval of enough cells with preserved viability.…”

Adipose-Derived Mesenchymal Stem Cells in the Use of Cartilage Tissue Engineering: The Need for a Rapid Isolation Procedure

“…Translating these techniques into the clinical setting will require the development of a rapid, sterile, one-step technique that could fit into a day surgery timeframe. To date, rapid isolation of bone marrow-derived MSCs [ 9 , 10 ] and their therapeutical potential has been studied [ 11 ], but an important barrier to adoption has been the low number of stem cells requiring a period of cell expansion in the laboratory. There is only one published study assessing a rapid isolation protocol (<30 minutes) for ADMSCs from abdominal lipoaspirate [ 12 ], but even this technique relies on a minimum of 24 hours for plastic adherence.…”