Our research presents a comprehensive and efficient method for producing kidney organoids from induced pluripotent stem cells, using suspension cultural conditions. We emphasize the importance of initial cell density and WNT agonist concentration during the inducing phase, which will educate new organoid researchers. Currently, it is difficult to obtain a single high-density cell line for iPSCs.

The batch impact was noticeable, particularly in various iPSC lines. Each batch's kidney organoids, may have a varied form and size. Several attempts are necessary to get the best initial cell density and concentration of CHIR for one iPSC line.

We have established that the initial differentiation of the intermediary method is important for successful kidney organoid formation. Our protocol describes in detail, how to choose the optimal cell densities and CHIR concentrations during the process because different iPSC lines showed variances in cell proliferation and differentiation ability. Researchers can quickly generate the specific kidney organoids from their iPSCs.

According to our protocol, by adjusting the cell density and the concentration of CHIR, it's undoubtedly beneficial for all our researchers to focus on their kidney disease. Our lab future research has to optimize the method of the protocol in disease models, regenerate new medicine and the drug screening. In recent five years, our lab will focus on the epigenetics and the cell topic drug development of various hereditary kidney disease, based on patient development, iPSC and the kidney organized disease models.