Serum-free non-toxic freezing solution for cryopreservation of human adipose tissue-derived mesenchymal stem cells.

OBJECTIVE: To develop a cost-effective, non-toxic and xeno-free freezing solution for the preservation of adipose tissue-derived stem cells (hADSC) with a long shelf-life. RESULTS: The potential of various hydrocolloids and organic osmolytes as cryoprotectants and individual components of phosphate buffered saline (PBS) as carrier media were evaluated to formulate a freezing solution for the cryopreservation of hADSCs. Among the hydrocolloids, the highest viability, 55 %, was achieved with post-thawed (after 48 h storage at -80 °C) hADSCs cryopreserved in 10 % (v/v) polyvinylpyrrolidone (PVP) using PBS as carrier media. 0.9 % NaCl was a superior carrier medium resulting an enhanced cell viability (70 %) when used in 10 % PVP than other components of PBS. A higher cell viability (81 %) was achieved when 10 % PVP/0.9 % NaCl was supplemented with 60 mM ectoin. The cryopreserved cells retained normal cytoskeletal distribution pattern and adipogenic and osteogenic differentiation ability during 14 and 21 days of incubation. CONCLUSION: A serum-free and non-toxic 10 % PVP/0.9 % NaCl/60 mM ectoin freezing solution was developed for cryopreservation of hADSC for application in tissue engineering and regenerative medicine.

Effects of spray parameters on skin tumour ablation volume during cryotherapy.

The main purpose of the study is to establish correlations for the ablation volume and the ice front as a function of the spray parameters. The ablation volume and the ice front depend upon the nozzle diameter, spraying distance and the freeze duration (spray parameters). The estimation of the ablation volume using the spray parameters shall be useful in surgical practice to ablate the different sizes of tumours. Liquid nitrogen spray cooling is carried out with 0.8 mm, 0.6 mm and 0.4 mm nozzle diameters. The spraying distance is maintained at 9 mm, 18 mm and 27 mm. The spray cooling is carried out for a single freeze-thaw cycle where freezing and thawing consist of 120 s and 130 s duration respectively. A two-dimensional heat flow equation with phase change is considered for the numerical study. The numerically calculated transient temperature (2 mm and 5 mm from the gel surface) and ice front values show confirmatory results with the experimentally measured data. Correlations are obtained to determine the ablation volume (- 50 °C and - 25 °C isothermal surfaces) and ice front (axial and lateral) with a goodness of fit [Formula: see text] 95%. The nozzle diameter has a greater impact on the ablation volume as compared to the spraying distance during 120 s of freezing. The nozzle diameter of 0.8 mm, 0.6 mm and 0.4 mm can be effectively used for cryotherapy with spraying distance up to 27 mm, 18 mm and 9 mm respectively.