Laboratory of Tissue Plasticity in Health and Disease, Cluster of Stem Cell and Developmental Biology, Department of Development and Regeneration, Leuven Stem Cell Institute, KU Leuven (University of Leuven).
Laboratory of Tissue Plasticity in Health and Disease, Cluster of Stem Cell and Developmental Biology, Department of Development and Regeneration, Leuven Stem Cell Institute, KU Leuven (University of Leuven);
J Vis Exp. 2022 Feb 25(180). doi: 10.3791/63431.
The pituitary is the master endocrine gland regulating key physiological processes, including body growth, metabolism, sexual maturation, reproduction, and stress response. More than a decade ago, stem cells were identified in the pituitary gland. However, despite the application of transgenic in vivo approaches, their phenotype, biology, and role remain unclear. To tackle this enigma, a new and innovative organoid in vitro model is developed to deeply unravel pituitary stem cell biology. Organoids represent 3D cell structures that, under defined culture conditions, self-develop from a tissue's (epithelial) stem cells and recapitulate multiple hallmarks of those stem cells and their tissue. It is shown here that mouse pituitary-derived organoids develop from the gland's stem cells and faithfully recapitulate their in vivo phenotypic and functional characteristics. Among others, they reproduce the activation state of the stem cells as in vivo occurring in response to transgenically inflicted local damage. The organoids are long-term expandable while robustly retaining their stemness phenotype. The new research model is highly valuable to decipher the stem cells' phenotype and behavior during key conditions of pituitary remodeling, ranging from neonatal maturation to aging-associated fading, and from healthy to diseased glands. Here, a detailed protocol is presented to establish mouse pituitary-derived organoids, which provide a powerful tool to dive into the yet enigmatic world of pituitary stem cells.
垂体是调节身体生长、代谢、性成熟、生殖和应激反应等关键生理过程的主要内分泌腺。十多年前，人们在垂体中发现了干细胞。然而，尽管采用了转基因活体方法，但其表型、生物学和作用仍不清楚。为了解决这个谜，开发了一种新的创新型体外类器官模型，以深入揭示垂体干细胞生物学。类器官是一种 3D 细胞结构，在特定的培养条件下，由组织（上皮）干细胞自我发育而来，并再现这些干细胞及其组织的多种特征。本文表明，源自小鼠垂体的类器官由腺体的干细胞发育而来，并忠实地再现了其体内表型和功能特征。其中，它们再现了干细胞的激活状态，这种激活状态类似于体内对转基因局部损伤的反应。类器官可长期扩增，同时保持其干细胞表型的强健。该新研究模型对于解析垂体重塑过程中关键条件下干细胞的表型和行为非常有价值，范围涵盖从新生儿成熟到与年龄相关的衰退，以及从健康到患病的腺体。这里呈现了建立小鼠垂体来源的类器官的详细方案，为深入研究神秘的垂体干细胞世界提供了有力工具。
