Designing a Low-Cost Spin-Drying Desiccation Technique Using 3D Printed Whirligig Model for Preservation of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells

In modern stem cell therapy, stem cells preservation is one of the major challenges. The present study aims to develop an advanced desiccation technique for the preservation of mesenchymal stem cells. Cryopreservation of human mesenchymal stem cells (MSCs) is one of the options for its long-term storage. The established techniques of preserving MSCs using cryopreservation aims to preserve cells at ultra-low temperatures using either liquid nitrogen or a deep freezer. However, limitations include cross-contamination, regular maintenance, the requirement of continuous liquid nitrogen supply, and electricity cost. A new technique of storing dried samples at ambient temperature has been developed utilizing desiccation. In this study, we desiccated the mesenchymal stem cells using a low-cost, power-free 3D printed whirligig model. It is an improved and effective method for preserving many different types of cells. We concluded that trehalose was introduced efficiently into the cells using fluid-phase endocytosis. We observed the effect of drying with trehalose solution at spinning speeds ranging from 2000 to 4000 rpm. To characterize the desiccation actuated cell death, we performed proliferation and viability analysis of post-desiccated MSCs by MTT assay. 60% of the cells were shown to survive this whirligig model based spin-drying in the presence of trehalose following immediate rehydration. Additionally, we explored apoptosis using the Annexin V-FITC detection kit and the presence of reactive oxygen species (ROS) by flow cytometry using DCFH-DA dye. In brief, this work provides a plausible methodology for preserving MSCs by desiccation.