Exosome Manufacturing Process
AnteAGE MDX® exosomes are isolated from the conditioned media of both bone marrow and umbilical cord mesenchymal stromal cells (BM-MSC & UC-MSC). From culturing the cells, collecting the conditioned media, purifying the exosomes, characterizing the exosomes, lyophilizing the finished product, quality control and regulatory check points, and filling and labelling vials all takes place in our state of the art exosome laboratory in Irvine, California.
Isolating the Exosomes
The exosomes are isolated from the BM-MSC and UC-MSC in the same way. Cell cultures are initiated in our class 5 ISO certified clean room, cells are seeded into flasks and cultured is xeno free medium. Cells are expanded into bioreactors to allow for the maximum number of cells from an initial culture. Cell culture media is replaced with a chemically defined low particle media for the production of stem cell conditioned media rich in exosomes.
Filtering & Filtration
Exosome rich conditioned media is chemically defined and contains no additional human or animal serum that can be a source of contaminating exosomes. Therefore all purified exosomes are known to have originated from the secreting stem cells. The exosome rich conditioned media is concentrated ~100x using tangential flow filtration (TFF); reducing the media volume from approximately 10L to 100mL. The membrane and pore size of the TFF system allows for small soluble proteins and water to be removed while retaining larger complex’s such as exosomes.
Purifying the Exosomes
The 100x concentrated conditioned media is then loaded onto a size exclusion chromatography (SEC) column to further purify the exosomes from the contaminating soluble proteins. Exosomes are purified in a salt buffer containing the cryoprotectant, Trehalose. Trehalose is important for the retention of vesicle structure following the lyophilization of each treatment vial.
Exosomes are characterized based on the ISEV minimal experimental requirements 2014 & 2018. Particle (exosome) concentration is determined by nanoparticle tracking analysis and exosomes are diluted and vials filled to contain 10 billion exosomes in each vial. Vials are rapidly frozen before lyophilization. Lyophilization occurs over 12 hours before septas are automatically compressed under vacuum.
Being in control of the complete manufacturing, quality testing and lot release allows AnteAGE® to produce a highly regulated, controlled, reproducible, and safe product for aesthetic use. Every lot undergoes rigorous testing prior to release including: sterility, endotoxin and exosome characterization and function in a number of different model systems. Prior to shipping, vials are stored in a temperature controlled environment and shipped with a cold pack to minimize any temperature fluctuation and ensuring a quality and reproducible product.