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. 2022 Jan 22;9(2):49. doi: 10.3390/bioengineering9020049

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Optimization protocol for the isolation of bone marrow cells. (A) Two cell preparations of bone marrow cells, density gradient separation (Ficoll^®) and IOR concentration step (CONC), were analysed at flowrate of 1 and 2 mL/min, giving an identical profile. (B) The two preparations were then tested adding the feature of the relaxation time of 3 min and run at a flowrate of 2 mL/min. This protocol gave a different profile, showing two main populations given by the two peaks. Bone marrow concentrated (CTRL) showed higher intensity in the first peak, which contained mainly RBCs, and also in the second one. Due to higher peak intensity not to mention avoidance of extra cell manipulation, the IOR concentration protocol was used for cell preparation. (C) One clinical sample was analysed to collect cells for different time interval (F1: 2–5 min; F2: 5–7 min; F3: 7–9 min) and the repetition of 4 analyses (RUN) showed reproducibility. Once cells were plated, only F1 cells attached to plate showed same morphology of CTRL (D) and showed ability to form CFU-F (E).