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Regen Med. 2019 May;14(4):279-293. doi: 10.2217/rme-2018-0043. Epub 2019 May 9.

Bench-to-bedside optimization of mesenchymal stem cell isolation, processing, and expansion for in vivo administration.

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United States Army Institute of Surgical Research, San Antonio, TX 78234, USA.
Oak Ridge Institute for Science & Education, Oak Ridge, TN 37831-0117, USA.
The Geneva Foundation, Tacoma, WA 98402, USA.


Aim: In this study, we aimed at identifying the optimal conditions for isolation, processing and expansion of mesenchymal stem cells (MSCs). Methods: Porcine bone marrow was obtained from either small- or large-volume bone marrow aspirate (BMA). Next, three BMA processing methods were compared. Finally, the best condition was selected from various culture parameters, including basal media, supplementation and seeding density. Results: Our results demonstrate that a small-volume BMA and direct plating yields significantly higher concentration of MSCs. Basal media supplementation with 10% platelet lysate and seeding density of 1000 cells/cm2 can generate large numbers of multipotent MSCs with augmented function and low population doublings. Conclusion: This work provides guidance for preparation of robust MSCs for future clinical trials.


bone marrow; bone marrow aspirate; colony-forming unit fibroblasts; fetal bovine serum; mesenchymal stem cells; platelet lysate; population doublings; seeding density


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