Fig. 4

Post-storage gene expression of stemness and surface markers of cADMSCs. A Gel results of genes. cADMSCs were suspended in various carrier solutions at 4 °C in a time-dependent manner and analyzed by RT-PCR for the expression of Nanog, Sox2, CD45, CD90, and CD105 followed by agarose gel electrophoresis. GAPDH was used as the housekeeping control gene. cADMSCs in all solutions did not express negative CD45. B, C, D, E Post-storage quantitative analysis of gene expression levels of cADMSCs. cADMSCs were suspended in various carrier solutions at 4 °C in a time-dependent manner. GAPDH was used as the housekeeping control gene. All mRNA data were normalized to the level of GAPDH and relative fold changes in expression level were measured. B The expression level of Nanog was significantly higher in HS at 1 h, which gradually decreased. Saline showed a significantly higher expression of Nanog at 12 h of storage. C A significantly high expression of Sox2 was found in 5% DS at 1 h and 12 h of storage. D The expression level of CD 90 up to 6 h was significantly higher in saline and PBS. E The expression level of CD105 at 12 h was significantly higher in PBS. Cells not stored in carrier solutions served as controls. The results are representative of three independent experiments. The bar graph represents the mean ± SD. cADMSCs - canine adipose-derived mesenchymal stem cells, GAPDH - Glyceraldehyde-3-Phosphate Dehydrogenase, Saline - 0.9% saline, PBS - Phosphate-buffered saline, 5% DS - 5% Dextrose solution, Hepa-Sal - Heparin in saline, HS - Hartmann’s solution, RT-PCR- Reverse transcription-polymerase chain reaction. SD - Standard deviation. (*P < 0.05, **P < 0.001, and ***P < 0.0001)