The effect of bone marrow aspiration strategy on the yield and quality of human mesenchymal stem cells

Abstract

Introduction Large inter-donor differences exist in human mesenchymal stem cell (hMSC) yield and the response of these cells to osteogenic stimuli. The source of these differences may be clinical differences in stem cell characteristics between individuals or the aspiration procedure itself.Methods From a total of 23 donors, we aimed to take 2 consecutive 10-mL aspirates from the same site in 17 donors and in 6 donors we aimed to take a 5-mL and a 20-mL aspirate. The aspiration was stopped either when the syringe was full or when no more bone marrow came through. Mononuclear cell yield (MNC), MSC yield, and differentiation capacity were analyzed for intra-donor and inter-donor variation. We analyzed the effect of the dilution with peripheral blood by drawing 20 mL at once.Results There was a high correlation between the first and second aspiration volumes, and aspirates with a volume of less than 8 mL showed a large variation in cellular yield. The second 10-mL aspirate, and also 20-mL aspirates, contained a lower concentration of nucleated cells and yielded lower numbers of mesenchymal stem cells. No effect of the aspiration procedure on the biological characteristics of the mesenchymal stem cells was seen.Conclusion We recommend collection volumes of bone marrow aspirates of at least 8 mL to reduce the risk of obtaining aspirates with low cell numbers. From the same site, a second aspiration or an aspirate of > 10 mL can be drawn without any loss of biological quality due to dilution with peripheral blood.

Clinical Aspects of Regenerative Medicine

“…Thus, a single BMA sample of 30-60 ml may not result in an optimal number of MSCs for therapeutic use. Fennema et al [73] suggested aspiration of ≥8 ml to reduce the chance of obtaining a BMA with low cell counts, without affecting the biologic characteristics of the MSCs. Generally, following centrifugation, 2-6 ml of BMACS is prepared for administration.…”

Quantification of biological variation in blood‐based therapy – a summary of a meta‐analysis to inform manufacturing in the clinic

“…Each has their own distinct advantages and foibles. Bone marrow has a higher risk to the donor that increases with age, requiring multiple aspirations from multiple sites to obtain optimal numbers 19 and can be painful. Peripheral blood is comparably more convenient for the donor as apheresis avoids anaesthesia and theatre.…”

Production of Clinical Grade Mesenchymal Stromal Cells

“…Fennema recommends to harvest at least 8 ml portions, as in lower volume the yield of nuclear cells and MSCs in unpredictable. Also, second portion of up to 10 ml may be taken from the same site without significant loss of quality due to dilution by peripheral blood (Fennema et al, 2009). On the other hand, in older work Muschler concludes that aspiration of more than 2 ml of bone blood results in smaller number of colony forming units -fibroblast (CFU-F) (Muschler et al, 1997).…”