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Cells 2018, 7(11), 190; https://doi.org/10.3390/cells7110190

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.
Received: 18 September 2018 / Revised: 12 October 2018 / Accepted: 23 October 2018 / Published: 30 October 2018
(This article belongs to the Section Stem Cells)
Full-Text   |   PDF [3011 KB, uploaded 30 October 2018]   |  

Abstract

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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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.

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