Slow-freezing cryopreservation of neural stem cell spheres with different diameters

Neural stem cells (NSCs) are of great value for clinical application and scientific research. The development of efficient cryopreservation protocols could significantly facilitate the storage and transportation for clinic applications. The objective of the present study is to improve the survival rate and viability of NSCs. Neural stem cells with three states of single-cell suspension, NSC spheres with diameters of 30–50 μm and 80–100 μm, were cryopreserved by slow-freezing method with the cryoprotective agent (CPA) of dimethyl sulfoxide (Me2SO), respectively. Then the post-thawing NSCs were tested for the survival rate and the differentiation ability. As a result, NSC spheres with diameter of 80–100 μm and Me2SO concentration of 8% achieve the survival rate of 82.9%, and the NSCs still sustain the multi-differentiation potentiality. These results indicated that both the subtle interaction among NSCs and sphere diameters may affect the survival rate together.