Stem Cells Out of the Bag: Characterization of Ex Vivo Expanded Mesenchymal Stromal Cells for Possible Clinical Use

Abstract

Aim: Mesenchymal stromal cells (MSC) are a promising tool for cellular therapy and regenerative medicine. One major difficulty in establishing a MSC expansion protocol is the large volume of bone marrow (BM) required. We studied whether cells trapped within a collection bag and filter system could be considered as a source of MSC. Results: From the 20 BM collection bag and filter systems, we recovered an average of 1.68 × 108 mononuclear cells, which is the equivalent to 60 ml of filtered BM. Mononuclear cells were expanded ex vivo to 17 × 106 MSC, with purity shown by a CD44+, CD105+, CD90+ and CD73+ immunophenotype, a reduction of 20% proliferating cells in a mixed lymphocyte reaction and also the ability of adipocyte differentiation. Conclusion: Long-term MSC cultures were established from the usually discarded BM collection bag and filter, maintaining an appropriate phenotype and function, being suitable for both investigation and clinical settings.

Turning Trash Into Treasure: A Simple Protocol for Human Mesenchymal Stromal Cell Isolation From Used Bone Marrow Collection Kits

“…However, this methodology requires skilled manual dexterity and entails a prolonged duration for the isolation of cells (Grisendi et al, 2010;Yamamoto et al, 2015;Zhu & Murthy, 2013). Alternatively, the evaluation of the bone marrow collection kits for Use premixed commercialized differentiation kits hMSC isolation has been a subject of high interest (Anasiz, 2018;Capelli et al, 2009;Lopes et al, 2020;Otsuru et al, 2013;Spohn et al, 2021). For clinical indications, harvested bone marrow from a suitable donor is pooled and filtered through the bone marrow collection kit to remove residual fat, bone chips, and clots (McDaniel et al, 2017).…”

Current Perspectives on Adult Mesenchymal Stromal Cell-Derived Extracellular Vesicles: Biological Features and Clinical Indications

“…The extensive investigations on BM-MSCs allowed the in-depth characterization of these cells with respect to alternative sources. Nonetheless, the isolation requires an invasive procedure with a risk of complications, and also the cellular yield from the BM decreases with the aging of the donor [ 103 , 104 ]. Specifically, Sacchetti et al provided evidence to confirm that BM-MSCs constitute a subset of perivascular/endothelial CD146+ cells with self-renewal capabilities in in vivo experiments [ 105 ].…”