Freeze-Dried and GMP-Compliant Pharmaceuticals Containing Exosomes for Acellular Mesenchymal Stromal Cell Immunomodulant Therapy

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.

Advances in Analysis of Biodistribution of Exosomes by Molecular Imaging

“…In an international effort to establish standards for exosome analysis, the Minimal Information for Studies of Extracellular Vesicles 2018 (MISEV 2018) was suggested through a series of publications [21,54,55]. Many studies also reported on the GMP production of exosomes for the development of therapeutics with suggested release criteria [45,50,[56][57][58][59][60][61]. The worldwide market for exosome-based therapy is expected to grow from 5 million USD in 2016 to 10.0 million USD in 2021, with a compound annual growth rate (CAGR) of 14.9% [62].…”

Characteristics and roles of extracellular vesicles released by epidermal keratinocytes

“…As a consequence, there is growing interest in the therapeutic application of EVs. The potential to manufacture clinical‐grade EVs containing therapeutic cargoes remains to be demonstrated, although current technologies on engineering and collecting EVs suggest this possibility is achievable . Clearly, further laboratory studies defining the clinical potential that EVs hold will be followed by the preclinical and clinical validations that are required to translate EV technologies into applications in the real world.…”

Steering the Clinical Translation of Delivery Systems for Drugs and Health Products

“…Important requirements for the development of a medicinal product are: To obtain a scalable and GMP-compliant production process and the stability and the shelf-life of the final product; ultrafiltration, as purifying process, and freeze-drying fit with this requirement. We combined these two techniques obtaining a ready-to-use powder named lyo-secretome [46,47]. EV integrity and morphology were not affected by the technological processes; product resulted as no cytotoxic and no hemolytic confirming its safety profile.…”

“…EV integrity and morphology were not affected by the technological processes; product resulted as no cytotoxic and no hemolytic confirming its safety profile. Efficacy of lyo-secretome was proven against oxidative stress-induced damages [2], as immunomodulant agent [47] and in a wound healing murine model. Furthermore, proteomic characterization of lyo-secretome revealed that it contains alpha1-antitripsin (AAT) with in vitro anti-elastase activity, paving the way for the use in lung degeneration AAT deficiency [48].…”