The R-Spondin proteins comprise a family of secreted proteins, known for their important roles in cell proliferation, differentiation and death, by inducing the Wnt pathway. Several studies have demonstrated the importance of RSPOs in regulation of a number of tissue-specific processes, namely: bone formation, skeletal muscle tissue development, proliferation of pancreatic β-cells and intestinal stem cells and even cancer. RSPO1 stands out among RSPOs molecules with respect to its potential therapeutic use, especially in the Regenerative Medicine field,due to its mitogenic activity in stem cells. Here, we generated a recombinant human RSPO1 （rhRSPO1） using the HEK293 cell line, obtaining a purified, characterized and biologically active protein product to be used in Cell Therapy.