SCAN: Novel cryopreservation strategies for cell-based therapies and pharmacological applications

Abstract

Effective preservation methodologies for cells and tissues are required to facilitate the supply of cell-based products for clinical, research and pharmacological applications. The standard cryopreservation protocol was developed for single cell suspensions and may not be able to assure the retention of high degrees of viability and function in complex systems such as stem cell colonies, monolayer cell cultures, 3-D aggregates or tissues. Improving currently used methodologies for cell preservation is a relevant subject of research taking into account the differences between such complex cell-systems and single cell suspensions regarding their responses to cooling, warming, and dehydration.

 The aim of this work is to understand and further characterize the effect of the cryopreservation process on the viability and function of cells to be frozen at different stages of differentiation. Alternative cryopreservation strategies to the standard protocol are being investigated based on:

i.    sample miniaturization and process automation;

ii.    cell entrapment within clinical grade, ultra-high viscosity alginate hydrogel;

iii.    use of specialized solutions designed to reduce the freeze-induced stress to the cells.

 Two cell lines, mouse neuroblastoma N2a and human colon adenocarcinoma Caco-2, are used as model systems due to their distinct structural and functional characteristics. Based on the knowledge acquired with these cell models, an improved cryopreservation methodology will be proposed and tested for stem cell colonies allowing the maintenance of the cell differentiation potential or, for the differentiation states thereafter, the cell specific functions after long-term storage.