Background The three-dimensional pituitary placode organoids hold extensive potential applications, yet current research in this field remains limited.
Objective To induce human induced pluripotent stem cells (hiPSCs) into pituitary placode organoids within an in vitro three-dimensional environment.
Methods hiPSCs were cultivated in suspension to form spheroids and gradually differentiated into pituitary placode organoids through supplement with a series of cell factors, agonists, or inhibitors. qPCR was employed to measure the mRNA expression levels of NANOG, SIX1, PITX1, and LHX3 in the organoids. Immunofluorescence staining was used to detect the expression of PITX1 and LHX3 in the organoids.
Results hiPSCs formed spheroids in vitro through suspension culture. As the culture time extended, the shape of the organoids changed from regular spherical to irregular. qPCR and immunofluorescence staining revealed successful expression of the pituitary placode markers PITX1 and LHX3 in the organoids at 30 and 40 days, respectively, indicating the successful construction of pituitary placode organoids.
Conclusion This study preliminarily establishes pituitary placode organoids, providing a laboratory-based model for pituitary development research, drug development, and regenerative medicine therapies.
DownLoad: