OPTIMIZATION OF EQUINE MESENCHYMAL STEM CELL CRYOPRESERVATION FOR CLINICAL USE
Abstract
Synovitis is the inflammation of the synovial membrane and is a common condition in horses. Synovitis can occur with many other conditions such as rheumatoid arthritis and can lead to joint degeneration if left untreated. The purpose of this study is to determine the optimal freezing condition to cryopreserve equine mesenchymal stem cells (MSCs) prior to use for synovitis control by intra-articular injection. In this study, MSCs were isolated from nine different horses and cryopreserved in the six conditions for each horse. After at least 24 hours, the cells were thawed and their cytoplasm was stained with standard CellTrace™ violet dye. The cells were then plated on two T25 flasks at a concentration of 10,000 cells/cm2 per condition and one colony formation assay at 10 cells/cm2 per condition. The remainder of the cells were set aside for flow cytometry as the baseline because they had not had time to undergo mitosis. The cell fluorescence from the dye was analyzed 24hr and 72hr post-plate respectively and the distribution of the dye was analyzed. With the colony formation assay, cell morphology, confluency, and data from the flow cytometer, a holistic idea of which condition produces the most viable cells post-thaw was hoped to be obtained. Upon a one-way analysis of variance (one-way ANOVA), it was found that there was no significant difference in cell viability post-thaw. This conclusion is further supported by statistical analysis of the CFU, cell counts, and post-thaw viabilities, which all produced large p-values indicating insignificance.
Citation
Ewing, Donald Cody (2014). OPTIMIZATION OF EQUINE MESENCHYMAL STEM CELL CRYOPRESERVATION FOR CLINICAL USE. Undergraduate Research Scholars Program. Available electronically from https : / /hdl .handle .net /1969 .1 /164415.