{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"Authorea, Inc."}],"indexed":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T16:38:46Z","timestamp":1748968726907,"version":"3.41.0"},"posted":{"date-parts":[[2022,12,21]]},"group-title":"Preprints","reference-count":0,"publisher":"Wiley","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2022,12,21]]},"abstract":"<jats:p id=\"p1\">The therapeutic effects of human mesenchymal stromal cells (MSC) have\nbeen attributed mostly to their paracrine activity, exerted through\nsmall-secreted extracellular vesicles (EVs) rather than their\nengraftment into injured tissues. Currently, the production of\nMSC-derived EVs (MSC-EVs) is performed in laborious static culture\nsystems with limited manufacturing capacity using serum-containing\nmedia. In this work, a serum-\/xenogeneic-free microcarrier-based culture\nsystem was successfully established for bone marrow-derived MSC\ncultivation and MSC-EV production using a 2 L-scale controlled stirred\ntank reactor (STR) operated under fed-batch (FB) or fed-batch combined\nwith continuous perfusion (FB\/CP). Overall, maximal cell numbers of\n(3.0\u00b10.12)\u00d710  and (5.3\u00b10.32)\u00d710 \nwere attained at days 8 and 12 for FB and FB\/CP cultures, respectively,\nand MSC(M) expanded under both conditions retained their\nimmunophenotype. MSC-EVs were identified in the conditioned medium\ncollected from all STR cultures by TEM, and EV protein markers were\nsuccessfully identified by WB analysis. Overall, no significant\ndifferences were observed between EVs isolated from MSC expanded in STR\noperated under the two feeding approaches. EV mean sizes of 163\u00b15.27 nm\nand 162\u00b14.44 nm (P&gt;0.05) and concentrations of\n(2.4\u00b10.35)x10  EVs\/mL and (3.0\u00b10.48)x10\n EVs\/mL (P&gt;0.05) were estimated by\nnanoparticle tracking analysis for FB and FB\/CP cultures, respectively.\nThe STR-based platform optimized herein represents a major contribution\ntowards the development of human MSC- and MSC-EV-based products as\npromising therapeutic agents for Regenerative Medicine settings.<\/jats:p>","DOI":"10.22541\/au.167160787.74320022\/v1","type":"posted-content","created":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T07:31:21Z","timestamp":1671607881000},"source":"Crossref","is-referenced-by-count":1,"title":["Optimized operation of a controlled stirred tank reactor system for the production of mesenchymal stromal cells and their extracellular vesicles"],"prefix":"10.22541","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5367-6528","authenticated-orcid":true,"given":"Ana","family":"Fernandes-Platzgummer","sequence":"first","affiliation":[{"name":"Universidade de Lisboa Instituto Superior Tecnico"}]},{"given":"Raquel","family":"Cunha","sequence":"additional","affiliation":[{"name":"Universidade de Lisboa Instituto Superior Tecnico"}]},{"given":"Sara","family":"Morini","sequence":"additional","affiliation":[{"name":"Universidade de Lisboa Instituto Superior Tecnico"}]},{"given":"Marta","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Universidade de Lisboa Instituto Superior Tecnico"}]},{"given":"Juan","family":"Moreno-Cid","sequence":"additional","affiliation":[{"name":"Bionet Servicios T\u00e9cnicos SL"}]},{"given":"Joaquim","family":"Cabral","sequence":"additional","affiliation":[{"name":"Universidade de Lisboa Instituto Superior Tecnico"}]},{"given":"Claudio Lobato da","family":"Silva","sequence":"additional","affiliation":[{"name":"Universidade de Lisboa Instituto Superior Tecnico"}]}],"member":"311","container-title":[],"original-title":[],"deposited":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T07:31:22Z","timestamp":1671607882000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.authorea.com\/users\/568256\/articles\/614196-optimized-operation-of-a-controlled-stirred-tank-reactor-system-for-the-production-of-mesenchymal-stromal-cells-and-their-extracellular-vesicles?commit=03e206e2bba52bfd9a5e199ab403557ec3070891"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,21]]},"references-count":0,"URL":"https:\/\/doi.org\/10.22541\/au.167160787.74320022\/v1","relation":{},"subject":[],"published":{"date-parts":[[2022,12,21]]},"subtype":"preprint"}}