Low scale production of advanced therapy medicinal products for limbal stem cell deficiency.

Abstract

Purpose : To develop a work environment to manufacture low scale advanced therapy medicinal products complying with GMP guidelines.

Methods : 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.

Results : 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.

Conclusions : 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.