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Alvaro Meana, Silvia Berisa, Manuel Chacon, Natalia Vazquez, MAIROBI PERSINAL MEDINA, Begoña Baamonde, Jose Alfonso, Luis Fernandez-Vega, Jesus Merayo-Lloves; Low scale production of advanced therapy medicinal products for limbal stem cell deficiency.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4104.
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© ARVO (1962-2015); The Authors (2016-present)
To develop a work environment to manufacture low scale advanced therapy medicinal products complying with GMP guidelines.
A low scale clean room environment was obtained using an isolator (ref: AISLAISO4-1800, TDI) located in a Grade C area. Isolator is equipped with a lateral SAS with independent UV sterilization system and double door lock; a gas outlet with electronically controlled valve and USB, HDMI and AC/DC connectors. Inside the isolator, a small portable CO2 incubator (Minicell NB203M, N-Biotek), a phase contrast microscope (EVOX XL Cell Imaging System, ThermoFisher) and a compact centrifuge (Spectrafuge 6C, Labnet) are installed.Limbal stem cell deficiency medicinal product is based on in vitro expanded limbal stem cells cultured on a platelet rich in growth factors (PRGF) fibrin membrane. Limbal explants are obtained from a local tissue bank and are cultured in DMEM:F12 supplemented with 10% human serum collected from human blood and 1% antibiotics. Limbal cells are cultured at 37C inside the isolator until confluence. Once confluence is reached, limbal cells are subcultured on a PRGF fibrin membrane (500.000 cells/membrane) and cultured for 3 to 5 days.
The developed work scheme is able to comply with current regulatory exigencies for advanced therapy medicinal products. Workspace under flow hood maintains an ISO 4 quality according to ISO 14644. All steps required for the production of an advanced therapy medicinal product in a cleanroom could be easily transferred to an isolator environment. Cultivation and subcultivation steps could be easily performed with disposable and sterile reagents and plastic ware with the only need of a centrifuge and CO2 incubator that were already located inside the isolator. Cell culture monitorization during the culture period could also be performed in the phase contrast microscope located inside the isolator.
A low scale work environment that complies with GMP guidelines could be a solution for the high-cost associated with cleanroom production of advanced therapy medicinal products. This work scheme could allow small laboratories to perform high-end therapies with a relatively low budget.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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