Biopreservation Best Practices for regenerative medicine GMP manufacturing & focus on optimized biopreservation media

Abstract

Cellular therapies are cell and tissue products sourced from biological materials that are employed as 'living drugs. ' Such 'living drugs' require specialized biological support, namely biopreservation, to maintain optimal recovery, viability and return to function post-preservation. To achieve successful biopreservation, optimization of a multitude of parameters, including cooling/thawing rates based on the biophysics of specific cell types, the temperature of storage and transportation, as well as biopreservation media are of paramount importance. In particular, the choice of biopreservation media is pivotal for clearing regulatory hurdles and facilitating commercialization. Traditional extracellular-like (isotonic), home-brew cocktails (which may contain serum) may not be compatible with a Good Manufacturing Practices (GMP) clinical manufacturing process. This article will address the challenges associated with maintaining the viable recovery and functionality of 'living drugs' , discuss the benefits and consequences of low-temperature biopreservation, outline Biopreservation Best Practices, and propose considerations for incorporating Biopreservation Best Practices into a GMP cell therapy product. Integration of Biopreservation Best Practices, beginning with the selection of optimized media, ensures that cells remain viable and functional throughout the cell product lifecycle, thereby optimizing manufacturing and clinical outcomes.

The Importance of Collection, Processing & Biopreservation Best Practices in Determining CAR-T Starting Material Quality

“…Best practice for shipping and storage of cell therapy material, whether it be starting material or the finished product, must include taking steps to protect cold chain integrity In addition to temperature optimization, stability of the starting material may be optimized with utilization of biopreservation media. An effective, well-designed biopreservation media mitigates risk by extending the shelf life of, and optimizing the quality of, cell therapy starting material [30,31]. The shelf life of freshly isolated leukapheresis products at room temperature is minimal; ranging anywhere from 30 minutes to a few hours, depending on the desired application [32].…”

Cryopreservation as a Key Element in the Successful Delivery of Cell-Based Therapies—A Review

“…An original isolate e.g., from biopsy or apheresis, can be stabilized and stored by cryopreservation until the appropriate time for further growth and/or manipulation to produce the final, therapeutic product. This final product can then, in turn, be cryopreserved and stored until the appropriate time and place for clinical delivery ( 8 , 30 , 31 ).…”

Cell therapies for trauma and critical care medicine: critical issues in translation for cellular and novel therapies in trauma and critical care

“…Traditional home‐brew reagent cocktails (including serum) utilized for biopreservation may be a point of risk within a cGMP clinical manufacturing process, and they may be suboptimal options in comparison to preformulated/prepackaged cGMP intracellular‐like formulations. Therefore, it would be worthwhile to consider best practices recommendations for integrating biopreservation methods within cGMP for cell therapy manufacturing, along with consideration to the overall quality and regulatory plan . Traditionally, biopreservation was generally a distinct task for the final product (either nonfrozen or frozen), not uncommonly an afterthought, and optimization might only be explored reactively from a negative output.…”

“…These methods leverage the decreased metabolism and reduced cellular degradation that result from low‐temperature conditions to preserve biologic integrity and function. However, cellular responses to low temperatures include detrimental outcomes, such as disruption of ionic and osmotic balance, accumulation of free radicals and reactive oxygen species, physical damage from ice formation, chemical damage from solute toxicity, and multiple mechanisms of cell death (necrosis, apoptosis, secondary necrosis) . Understanding these outcomes and the role of delayed‐onset cell death in each cell manufacturing product is critical, as measurement and interpretation of viability can be complex.…”

“…However, cellular responses to low temperatures include detrimental outcomes, such as disruption of ionic and osmotic balance, accumulation of free radicals and reactive oxygen species, physical damage from ice formation, chemical damage from solute toxicity, and multiple mechanisms of cell death (necrosis, apoptosis, secondary necrosis) . Understanding these outcomes and the role of delayed‐onset cell death in each cell manufacturing product is critical, as measurement and interpretation of viability can be complex. Often the assessment involves consideration of “perceived viability” immediately after preservation versus long‐term “true viability” of the cell product, as postpreservation cellular mechanisms may be slower to initiate, manifest, and resolve.…”