Aby Mathew

Improved Hypothermic Preservation of Human Renal Cells Through Suppression of Both Apoptosis and Necrosis

A new platform of hypothermic solutions, the HypoThermosol® (HTS) series, has been developed for the improved hypothermic storage of cells, tissues, and organs. Cells and tissues cold-stored in HTS-FRS demonstrate improved viability when returned to normothermic temperatures in comparison with the parent solution, HTS-BASE, or University of Wisconsin (UW) solution (UW-ViaSpan®). While our group and others have implicated apoptosis as a major player in cell death initiated by extended hypothermic storage, it has been unclear if the improved performance of HTS-FRS as a hypothermic storage solution is due to its ability to inhibit apoptosis. Data reported herein show that human renal cells hypothermically stored in renal cell culture medium, HTS-FRS, HTS-BASE, or UW solution demonstrated improved survival in HTS-FRS. Following 5 days of hypothermic preservation and 1 day of recovery at 37°C, cells preserved in HTS-FRS exhibit 75% metabolic activity, whereas cells stored in HTS-BASE, UW, or culture media demo...

C-38: Biopreservation considerations for GMP bioprocessing, clinical development, and delivery

Often in cell and tissue processing, there may exist a gap between biopreservation method optimization from a cryobiology perspective and the process development that results in the cryopreserved or non-frozen cell/tissue product. This presentation will discuss Best Practices recommendations for integrating biopreservation methods within Good Manufacturing Practices (GMP), share lessons learned from cell therapy manufacturing – including some positive and negative examples of biopreservation issues, and suggestions for integration of biopreservation methods within biobanking and Regenerative Medicine with consideration to the Quality and Regulatory footprint. Disclosure: Author was or is employed by BioLife Solutions, Inc.

052 Considerations for development of GMP Biopreservation Solutions for Regenerative Medicine

Cellular therapies and regenerative medicine utilize cell and tissue products sourced from blood, bone marrow, and various tissues. The clinical and commercial success of these products is potentially impacted by stability limitations, which include transport of the source material and biopreservation of the final cell or tissue product (either frozen or non-frozen). Traditional isotonic/extracellular-like home-brew reagent cocktails (including serum) utilized for biopreservation are an area of risk within a GMP (Good Manufacturing Practices) clinical manufacturing process. This discussion will address points of Quality/Regulatory concern for qualification of biopreservation reagents within GMP systems. Topics include transportation and storage of source material and final dose, intermediate manufacturing process hold steps, and evaluation, selection, and qualification of ancillary and excipient reagents. A case study sharing the development of GMP cryopreservation media into clinical applications will be discussed. Source of funding: None declared. Conflict of interest: None declared. amathew@biolifesolutions.com

Transplantation Diagnostics: A Preliminary Analysis Using Protein Microarray to Determine Kidney Status Prior To and Following Implantation

The need for enhanced transplant diagnostics is reflected in the numbers of unused organs as indicated by the United Network for Organ Sharing. We hypothesize that the number and quality of organs for transplant could be greatly enhanced with the inclusion of proteomic analysis. To test this prospect, we utilized the surface enhanced laser desorption/ionization time-of-flight system (SELDI-TOF), to identify phenomic fingerprints of porcine kidney flush solution collected following cold preservation and post-transplant protein lysates from human kidney transplant urine samples. Effluent flush solution from porcine kidneys statically stored under hypothermic conditions for 6 days yielded novel protein peaks in the 7350-15,950-Da range from day 1 to day 3 of storage, and peak intensification from day 4 to day 6. Comparison of donor (pre-transplant) and recipient (post-transplant) cellular protein extracts collected from urine samples demonstrated several peaks of differing intensities between 11,997 and 64,0...