Abstract: Objective To investigate whether the gelatin microcryogels can maintain biological characteristics of human mesenchymal stem cells from adipose tissue (ADSCs) and enhance the stemness of ADSCs when they are co-cultured in vitro. Method Calcein-AM and PI staining were conducted to test cell viability of ADSCs plated in gelatin microcryogels. Cell titer-blue assay was used to examine the cellular proliferating capacity. Real-time PCR was used to evaluate the expression level of the stemness genes OCT4、Nanog、SOX2. Adipogenic and osteogenic differentiation were induced and compared in ADSCs plated in gelatin microcryogels and in conventional environment. Results ADSCs were biologically active in the 3D scaffolds of gelatin microcryogels. Proliferation rate was not influenced. Stemness genes expression was up-regulated. ADSCs plated in gelatin microcryogels still had osteogenic and adipogenic differentiation ability, but the ralated genes expression levels were lower in comparison with ADSCs in conventional inducing condition. Conclusion Gelatin microcryogels can provide proper microenvironmental factors for ADSCs stemness maintenance, so as to exert the long-term application value in the diseases treated with stem cell by a minimally-invasive delivery method.

Key words: Human mesenchymal stem cells from adipose tissue(ADSCs), Gelatin microcryogels