Container system for enabling commercial production of cryopreserved cell therapy products
Abstract
Aim: The expansion of cellular therapeutics will require large-scale manufacturing processes to expand and package cell products, which may not be feasible with current blood-banking bag technology. This study investigated the potential for freezing, storing and shipping cell therapy products using novel pharmaceutical-grade Crystal Zenith® (CZ) plastic vials. Materials & methods: CZ vials (0.5, 5 and 30 ml volume) with several closure systems were filled with mesenchymal stem cells and stored at either -85 or -196°C for 6 months. Vials were tested for their ability to maintain cell viability, proliferative and differentiation capacity, as well as durability and integrity utilizing a 1-m drop test. As controls, 2 ml polypropylene vials were investigated under the same conditions. Results: Post-thaw viability utilizing a dye exclusion assay was over 95% in all samples. Stored cells exhibited rapid recovery 2 h post-thaw and cultures were approximately 70% confluent within 5–7 days, consistent with nonfrozen controls and indicative of functional recovery. Doubling times were consistent over all vials. The doubling rate for cells from CZ vials were 2.14 ± 0.83 days (1 week), 1.84 ± 0.68 days (1 month) and 1.79 ± 0.71 days (6 months), which were not significantly different compared with frozen and fresh controls. Cells recovered from the vials exhibited trilineage differentiation consistent with controls. As part of vial integrity via drop testing, no evidence of external damage was found on vial surfaces or on closure systems. Furthermore, the filled vials stored for 6 months were tested for container closure integrity. Vials removed from freezer conditions were transported to the test laboratory on dry ice and tested using pharmaceutical packaging tests, including dye ingress and microbial challenge. The results of all stoppered vials indicated container closure integrity with no failures. Conclusion: Pharmaceutical-grade plastic CZ vials, which are commercially used to package pharmaceutical products, are suitable for low-temperature storage and transport of mesenchymal stem cells, and are a scalable container system for commercial manufacturing and fill-finish operation of cell therapy products.
Bibliography
- 1 Kirouac DC, Zandstra PW: The systematic production of cells for cell therapies. Cell Stem Cell3,369–381 (2008).
- 2 Akers MJ, Nail SL, Saffell-Clemmer W: Top ten topics in parenteral science and technology. PDA J. Sci. Technol.61(5),337–361 (2007).
- 3 Eakins MN: New plastics for old vials. Bioprocess Int.3(6),52–58 (2005).
- 4 Qadry SS, Roshdy TH, Char H, Del Terzo S, Tarantino R, Moschera J: Evaluation of CZ-resin vials for packaging protein-based parenteral formulations. Int. J. Pharm.252,207–212 (2003).
- 5 Esfandiary R, Joshi SB, Vilivalam V, Middaugh CR: Characterization of protein aggregation and adsorption on prefillable syringe surfaces. Presented at: AAPS National Biotechnology Conference. Toronto, Canada, 22–25 June 2008.
- 6 Waxman L, Vilivalam V: Development of analytical techniques to determine protein adsorption on sterilized parenteral packaging containers and stoppers. Presented at: AAPS National Biotechnology Conference. Seattle, WA, USA, 21–24 June 2009.
- 7 Shi S, Robey PG, Gronthos S: Comparison of human dental pulp and bone marrow stromal stem cells by cDNA microarray analysis. Bone29,532–539 (2001).
- 8 Perry BC, Zhou D, Xiaohua W et al.: Collection, cryopreservation and characterization of human dental pulp derived mesenchymal stem cells for banking and clinical use. Tissue Eng. Part C14,149–156 (2008).
- 9 Seo BM, Miura M, Sonoyama W, Coppe C, Stanyon R, Shi S: Recovery of stem cells from cryopreserved periodontal ligament. J. Dent. Res.841,907–912 (2005).
- 10 Zhang W, Walboomers XF, Shi S, Fan M, Jansen JA: Multilineage differentiation potential of stem cells derived from human dental pulp after cryopreservation. Tissue Eng.12,2813–2823 (2006).
- 11 Woods EJ, Perry BC, Hockema JJ, Larson L, Zhou W, Goebel WS: Optimized cryopreservation method for human dental pulp derived stem cells and their tissues of origin for banking and clinical use. Cryobiology59(2),150–157 (2009).
- 12 Dominici M, Le Blanc K, Mueller I et al.: Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy8,315–317 (2006).
- 13 Freshney RI: Culture of Animal Cells: A Manual of Basic Technique. John Wiley & Sons, Inc., NY, USA (2000).
- 14 Khuu HM, Cowley H, David-Ocampo V et al.: Catastrophic failures of freezing bags for cellular therapy products: description, cause, and consequences. Cytotherapy4(6),539–549 (2002).
- 101 Daikyo Resin CZ Technical Report, West Pharmaceutical Services (2002) www.westpharma.com/ap/en/products/Pages/CrystalZenithRU.aspx
