Alginate-Encapsulation for the Improved Hypothermic Preservation of Human Adipose-Derived Stem Cells

Abstract

Despite considerable progress within the cell therapy industry, unmet bioprocessing and logistical challenges associated with the storage and distribution of cells between sites of manufacture and the clinic exist. We examined whether hypothermic (4°C-23°C) preservation of human adipose-derived stem cells could be improved through their encapsulation in 1.2% calcium alginate. Alginate encapsulation improved the recovery of viable cells after 72 hours of storage. Viable cell recovery was highly temperaturedependent, with an optimum temperature of 15°C. At this temperature, alginate encapsulation preserved the ability for recovered cells to attach to tissue culture plastic on rewarming, further increasing its effect on total cell recovery. On attachment, the cells were phenotypically normal, displayed normal growth kinetics, and maintained their capacity for trilineage differentiation. The number of cells encapsulated (up to 2 3 10 6 cells per milliliter) did not affect viable cell recovery nor did storage of encapsulated cells in a xenofree, serum-free,current Good Manufacturing Practice-grade medium. We present a simple, low-cost system capable of enhancing the preservation of human adipose-derived stem cells stored at hypothermic temperatures, while maintaining their normal function. The storage of cells in this manner has great potential for extending the time windows for quality assurance and efficacy testing, distribution between the sites of manufacture and the clinic, and reducing the wastage associated with the limited shelf life of cells stored in their liquid state. STEM CELLS TRANSLATIONAL MEDICINE 2016;5:339-349 SIGNIFICANCEDespite considerable advancement in the clinical application of cell-based therapies, major logistical challenges exist throughout the cell therapy supply chain associated with the storage and distribution of cells between the sites of manufacture and the clinic. A simple, low-cost system capable of preserving the viability and functionality of human adipose-derived stem cells (a cell with substantial clinical interest) at hypothermic temperatures (0°C-32°C) is presented. Such a system has considerable potential for extending the shelf life of cell therapy products at multiple stages throughout the cell therapy supply chain.

A novel antifibrotic strategy utilizing conditioned media obtained from miR-150-transfected adipose-derived stem cells: validation of an animal model of liver fibrosis

“…Isotype controls were performed with antibodies against IgG. In addition, the adipogenic, osteogenic, and chondrogenic differentiation ability of the naive and transfected ASCs was determined as previously described 32,33 .…”

Synergistic Development of Biochips and Cell Preservation Methodologies: a Tale of Converging Technologies

“…Due to the fact that no ice forms during the process, the effects of cooling are often reversible. Hypothermia can be also used for short-term cell preservation on the order of hours to days [19–21]. However, long-term preservation of cells has not been achieved using this approach.…”

The role of biopreservation in cell and gene therapy bioprocessing

“…Cell viability post-preservation was found to be 80% and 74% for the hMSCs and mESCs respectively [47]. Similarly, Swioklo et al (2016) showed human adipose-derived stem cells could be encapsulated and stored at a range of temperatures, from 4-23C, although 15C was found to be the optimal [48]. The disadvantage to this method however is the need for a cell retrieval step which increases process complexity and will likely need to be undertaken at-or nearclinic.…”