Elia Bari1, Sara Perteghella1,2, Laura Catenacci1, Marzio Sorlini2,3, Stefania Croce4, Melissa Mantelli5, Maria A Avanzini5, Milena Sorrenti1, Maria L Torre1,2. 1. Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy. 2. PharmaExceed srl, 27100 Pavia, Italy. 3. SUPSI, Department of Innovative Technologies, University of Applied Sciences & Arts of Southern Switzerland, Via Pobiette 11, 6928 Manno, Switzerland. 4. Department of Clinical, Surgical, Diagnostic & Pediatric Sciences, University of Pavia, 27100 Pavia, Italy. 5. Fondazione IRCCS Policlinico S. Matteo, Immunology & Transplantation Laboratory/Pediatric Surgery, Cell Factory & Regenerative Medicine Research Center, 27100 Pavia, Italy.
Abstract
AIM: To validate the use of ultrafiltration (UF) as an alternative applicable industrial method to replace ultracentrifugation (UC) in the purification of mesenchymal stromal cell (MSC)-secretome. MATERIALS & METHODS: Pharmaceutical formulations containing secretome and/or extracellular vesicles were extracted from adipose-MSCs and bone marrow-MSCs by combining UF or UC with lyophilization. RESULTS & CONCLUSION: UF led to higher protein, lipid, cytokine and exosomes yields compared with UC. The isolation procedure and cell source influenced immunomodulatory activity, which was in vitro evaluated by inhibition of phytohemagglutinin-activated peripheral blood mononuclear cell proliferation, and by modulation of IL-10, IFN-γ and IL-6. A secretome dosage was identified to obtain the same immunomodulatory activity of MSCs, paving the way for cell-free therapy.
AIM: To validate the use of ultrafiltration (UF) as an alternative applicable industrial method to replace ultracentrifugation (UC) in the purification of mesenchymal stromal cell (MSC)-secretome. MATERIALS & METHODS: Pharmaceutical formulations containing secretome and/or extracellular vesicles were extracted from adipose-MSCs and bone marrow-MSCs by combining UF or UC with lyophilization. RESULTS & CONCLUSION: UF led to higher protein, lipid, cytokine and exosomes yields compared with UC. The isolation procedure and cell source influenced immunomodulatory activity, which was in vitro evaluated by inhibition of phytohemagglutinin-activated peripheral blood mononuclear cell proliferation, and by modulation of IL-10, IFN-γ and IL-6. A secretome dosage was identified to obtain the same immunomodulatory activity of MSCs, paving the way for cell-free therapy.