Enabling clinical trials in an academic GMP setting through use of closed, semi-automated manufacturing of allogeneic amniotic epithelial cells

• Published in: Cytotherapy (Oxford, England) Vol. 21; no. 5; p. S37
• Main Authors: Lim, R, Li, A, Kusuma, G, Chan, S, McPhee, G, Fitzpatrick, I, Wilson, S, James, D
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01-05-2019
• Subjects: Advanced Basic Science; Other
• ISSN: 1465-3249; 1477-2566
• DOI: 10.1016/j.jcyt.2019.03.370

Background & AimOne of the major considerations with regards to the design, usage and ultimately, viability of academic GMP facilities for clinical trials is cost effectiveness. This is particularly the case when clinical trials are supported by scant research funds. In the case of the Monash Health Translation Precinct's Cell Therapy Platform, the Biospherix Xvivo isolator was determined to be a cost-effective method to rapidly create a Class A environment within a Class D room to support manual, open processing. This approach was sufficient for the needs of a small Phase 1, single low dose study where allogeneic amniotic epithelial cells (hAEC) were isolated, cryopreserved and formulated extremely premature babies with chronic lung disease (n=6). For that trial, only 5 placentas were processed. Since the successful completion of this first-in-human trial, the Monash Health Translation Precinct has seen an explosion of hAEC clinical trials including dose-escalation studies in acute stroke, liver cirrhosis, and Crohn's disease. The projected demand on our academic GMP facility is estimated at over 250 processes per year for hAEC isolation alone. Given the labour-intensive nature of hAEC isolation (approx. 4 hours per isolation), and competing demands on the isolator at our academic GMP facility, there is a clear need to move towards closed, automated processing.Methods, Results & ConclusionAccordingly, we applied the use of the Rotea TM (Scinogy Pty Ltd), a benchtop counterflow centrifugal device and single-use kit, to the post-trypsinisation steps of the process. Once separated from the underlying chorion, the amniotic membrane is trimmed into 4 × 4cm pieces and exposed to 0.25% Trypsin-EDTA for 1 hour at 37°C. This is then quenched with 2% (v/v) human serum albumin in saline. The subsequent steps involving red blood cell removal and repeated washing steps (∼ 90 mins in the isolator), were replaced by the use of the Rotea TM (∼10 mins). Furthermore, the Rotea TM steps could be performed outside of the isolator, in a Class D environment with no significant loss in total live cells recovered. This is critical, as it allows staff to clean the isolator and prepare for the entry of incoming material, thus allowing for a level of scalability and flexibility in our current academic GMP manufacturing workflow that could never previously be achieved. Comparing isolation processes alone, this semi-automation will approximately triple the efficiency of our facility due to its enclosed, single-use system.