INVESTIGATION OF HUMAN CULTURED CELLS VIABILITY IN SUSPENSION

The success of cellular therapy is directly related to the viability of transplanted cells. In some cases the cells may be injected as suspension. However, the optimal conditions for maintain of the cell viability during graft preparation and cell suspension storage have not yet been established. Purpose of this study is to investigate the optimal conditions for storage of suspensions of human submandibular salivary gland cells, differentiated submandibular salivary gland cells and dermal fibroblasts in a physiologically compatible solutions. Standard methods for cell isolation and cultivation were used. Counting was performed on an automatic cell counter BioRad, cell viability was assessed by staining with 4% Trypan blue. As biocompatible solutions we tested phosphate-buffered saline, saline solution for injection and a 2% solution of human albumin in phosphate-buffered saline. It was found that the tested human cells retain viability in suspension in all solutions for at least 24 hours at +4°C and +25°C. Highest salivary gland cells viability (more 50%) was observed in phosphate buffered saline at both storage temperatures. However, the salivary gland cells better maintain adhesive and proliferative properties after 24 hours of incubation at +4°C. The study of fibroblasts shows that in saline solution these cells are preserved as a single cell suspension and hardly lose viability during 30 hours storage at +4°C. Addition of 2% albumin reduces the viability of fibroblasts. Based on our studies we recommend to store and transport the human submandibular salivary gland cells in phosphate buffered saline at +4°C; human fibroblasts — in saline solution at +4°C.

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