Journal Article

Development and Validation of a Fully GMP-Compliant Process for Manufacturing Stromal Vascular Fraction: A Cost-Effective Alternative to Automated Methods

Vogtensperger, Marie, Magalon, Jérémy, Dumoulin, Chloé, Velier, Mélanie, Arnaud, Laurent, Aboudou, Houssein, Lyonnet, Luc, François, Pauline, Simoncini, Stéphanie, Dignat-George, Françoise, Guillet, Benjamin, Grimaud, Fanny, Sabatier, Florence, Veran, Julie, Bertrand, Baptiste, Giraudo, Laurent

2020

Overview

The therapeutic use of adipose-derived stromal vascular fraction (SVF) is expanding in multiple pathologies. Various processes have been proposed for manufacturing SVF but they must be revisited based on advanced therapy medicinal product (ATMP) regulations. We report here the development and validation of a fully good manufacturing practices (GMP)-compliant protocol for the isolation of SVF. Adipose tissue was collected from healthy volunteers undergoing lipoaspiration. The optimal conditions of collagenase digestion and washing were determined based on measurements of SVF cell viability, yield recovery, and cell subset distribution. Comparability of the SVF obtained using the newly developed manufacturing process (n = 6) and the Celution-based automated method (n = 33), used as a reference, was established using inter-donor analyses. Characteristics of SVF (n = 5) generated using both manufacturing protocols were analyzed for an intra-donor comparison. In addition, these comparisons also included the determination of colony-forming unit fibroblast frequency, in vitro angiogenic activity, and in vivo regenerative effects in a mouse ischemic cutaneous wound model. We successfully developed a process for the generation of SVF presenting higher cell viability and yield recovery compared to the Celution device-based protocol. Characteristics of the SVF including phenotype, capacity for angiogenesis, and wound-healing promotion attested to the comparability of the two manufacturing processes. We validated an optimized non-automated process that should allow for a GMP-compliant, more affordable, and reduced-cost strategy to exploit the potential of SVF-based regenerative therapies.

• Publisher: MDPI AG,MDPI
• Subject: Adipose tissue / Adipose Tissue - blood supply / Adipose Tissue - cytology / Angiogenesis / Animals / Automation / Biological products / Cell Culture Techniques - economics / Cell Culture Techniques - methods / cell therapy / Cell viability / Collagen / Collagenase / Collagenases - metabolism / Comparative analysis / Cost-Benefit Analysis / Disease Models, Animal / Enzymes / Female / GMP-compliant manufacturing / Good Manufacturing Practice / Hospitals / Human health and pathology / Humans / Ischemia / Ischemia - pathology / Kinetics / Life Sciences / Manufacturing / Medical equipment / Methods / Mice, Nude / Neovascularization, Physiologic / Phenotypes / Physiological aspects / Plastic surgery / Production processes / Quality control / Stromal Cells - cytology / stromal vascular fraction / Substrate Specificity / Wound healing