Overview

This study investigated the efficacy of recombinant Sartorius Research Use Only (RUO) Cytokines and Growth Factors in generating organoids, improving expansion, and allowing researchers precise control over culture conditions. We demonstrate their capacity to both significantly enhance and sustain organoid growth for extended periods, and successfully produce desired cell types during differentiation of induced pluripotent stem cells (iPSCs).

Introduction

The shift to complex models such as organoids due to their physiological features, accurate organ architecture, and cell type interactions, alongside inclusion of iPSCs allows a personalized approach to studying pathologies. Together, these improve the accuracy of pre-clinical studies and reduce reliance on animal models. However, the culture conditions for these systems require a fine balance of components including growth factors to maintain health and expansion capacity.

Methods

Culture medium containing Sartorius RUO Growth Factors EGF, FGF10, HGF, and R-spondin1 was used to expand mouse and human hepatic organoids. Organoid cultures were automatically monitored and metrics including organoid area, count, and darkness were tracked using the Incucyte® Organoid Analysis Software Module. Human iPSCs were differentiated to mature hepatocytes using precise timing and concentrations of growth factors. The iQue® Advanced Flow Cytometry Platform was used for marker analysis.

Results

Organoid growth was improved over both media without growth factors and commercially available organoid medium when Sartorius RUO Growth Factors were present. Mouse hepatic organoids also demonstrated concentration-dependent growth curves in response to varied concentrations of R-spondin1. Human iPSCs were successfully differentiated to mature hepatocytes as confirmed by increased expression of hepatic markers and decreased pluripotency markers.

Conclusion

Sartorius RUO Growth Factors and Cytokines combined with the Incucyte® Live-Cell Analysis System and the iQue® Advanced Flow Cytometer allow precise control and monitoring of organoid culture. Together they form a comprehensive workflow from ensuring the quality of initial iPSCs, confirming vital checkpoints during differentiation, to characterisation of mature organoids. This allows researchers confidence in their processes and has further applications in drug discovery and toxicity studies.