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Pilot Production of Mesenchymal Stem/Stromal Freeze-Dried Secretome for Cell-Free Regenerative Nanomedicine: A Validated GMP-Compliant Process

1
Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
2
PharmaExceed srl, 27100 Pavia, Italy
3
Institute for Biomedical Technologies, F.lli Cervi 93, 20090 Segrate, Milan, Italy
4
Department of Innovative Technologies, University of Applied Sciences and Arts of Southern Switzerland, SUPSI, Via Pobiette 11, 6928 Manno, Switzerland
5
Tissue Therapy Unit, ASST Niguarda Hospital, Piazza Ospedale Maggiore 3, 20162 Milan, Italy
*
Author to whom correspondence should be addressed.
Cells 2018, 7(11), 190; https://doi.org/10.3390/cells7110190
Received: 18 September 2018 / Revised: 12 October 2018 / Accepted: 23 October 2018 / Published: 30 October 2018
(This article belongs to the Section Stem Cells)
In this paper, a pilot production process for mesenchymal stem/stromal freeze-dried secretome was performed in a validated good manufacturing practice (GMP)-compliant cell factory. Secretome was purified from culture supernatants by ultrafiltration, added to cryoprotectant, lyophilized and characterized. We obtained a freeze-dried, “ready-off-the-shelf” and free soluble powder containing extracellular vesicles and proteins. In the freeze-dried product, a not-aggregated population of extracellular vesicles was detected by nanoparticle tracking analysis; Fourier transform infrared spectra showed the simultaneous presence of protein and lipids, while differential scanning calorimetry demonstrated that lyophilization process successfully occurred. A proteomic characterization allowed the identification of proteins involved in immune response, response to stress, cytoskeleton and metabolism. Moreover, the product was not cytotoxic up to concentrations of 25 mg/mL (on human fibroblasts, chondrocytes and nucleus pulposus cells by MTT assay) and was blood compatible up to 150 mg/mL. Finally, at concentrations between 5 and 50 mg/mL, freeze-dried secretome showed to in vitro counteract the oxidative stress damage induced by H2O2 on nucleus pulposus cells by MTT assay. View Full-Text
Keywords: exosome; microvesicles; ultrafiltration; freeze-drying; secretome; mesenchymal stem cells exosome; microvesicles; ultrafiltration; freeze-drying; secretome; mesenchymal stem cells
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MDPI and ACS Style

Bari, E.; Perteghella, S.; Di Silvestre, D.; Sorlini, M.; Catenacci, L.; Sorrenti, M.; Marrubini, G.; Rossi, R.; Tripodo, G.; Mauri, P.; Marazzi, M.; Torre, M.L. Pilot Production of Mesenchymal Stem/Stromal Freeze-Dried Secretome for Cell-Free Regenerative Nanomedicine: A Validated GMP-Compliant Process. Cells 2018, 7, 190. https://doi.org/10.3390/cells7110190

AMA Style

Bari E, Perteghella S, Di Silvestre D, Sorlini M, Catenacci L, Sorrenti M, Marrubini G, Rossi R, Tripodo G, Mauri P, Marazzi M, Torre ML. Pilot Production of Mesenchymal Stem/Stromal Freeze-Dried Secretome for Cell-Free Regenerative Nanomedicine: A Validated GMP-Compliant Process. Cells. 2018; 7(11):190. https://doi.org/10.3390/cells7110190

Chicago/Turabian Style

Bari, Elia; Perteghella, Sara; Di Silvestre, Dario; Sorlini, Marzio; Catenacci, Laura; Sorrenti, Milena; Marrubini, Giorgio; Rossi, Rossana; Tripodo, Giuseppe; Mauri, Pierluigi; Marazzi, Mario; Torre, Maria L. 2018. "Pilot Production of Mesenchymal Stem/Stromal Freeze-Dried Secretome for Cell-Free Regenerative Nanomedicine: A Validated GMP-Compliant Process" Cells 7, no. 11: 190. https://doi.org/10.3390/cells7110190

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