Protocol development and integrating automating into preservation protocols

The development of preservation protocols has typically involved selecting a composition and a cooling rate and then measuring the resulting post thaw viability.  This process is repeated until an acceptable post thaw recovery is determined.  This approach is time consuming and costly.  Our approach is to use Differential Evolution to efficiently optimize composition and cooling rate for a given cell type.  This approach has enabled us to reduce the number of experiments required for optimization significantly. 

Tissues contain a variety of cell types.  Effective preservation protocols must result in high levels of post thaw recovery for all of the cell types present in a given tissue or mixed cell population.  We are presently applying Differential Evolution with Multiple Objects to efficiently optimize the preservation of multiple cell types.

Preservation protocols are typically labor intensive and there has been little integration of automation into the preservation process.  An active area of research involves the integration of robotics/automation into the preservation process such as retrieval of samples and controlled ice nucleation.  We have demonstrated the ability to use robotic cell culture and manipulation to at both the beginning of the preservation process as well as thawing and post thaw culture of cells.  We propose to extend the use of automation to process using high throughput techniques, a large number of biological systems and thereby enhance the throughput for specific systems such as drosophila melanogaster, which require preservation of > 150,000 different lines. 


Differential evolution optimization for pluripotent stem cells



Cryopreservation of Human iPS Cell Aggregates in a DMSO-Free Solution—An Optimization and Comparative Study - Rui Li, Kathlyn Hornberger, James R. Dutton, and Allison Hubel



 Programmable high-throughput screening robotic system and algorithm validation (Tecan)




Comparison of algorithm based optimization (right) compared to a full high-throughput screen of the parameter space to optimize preservation of Jurkat cells for a given combination of osmolytes


Algorithm‐Driven Protocol Optimization (Chapter 8) - Allison Hubel