Department of Plastic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Methods Mol Biol. 2024;2783:235-262. doi: 10.1007/978-1-0716-3762-3_16.
Advances in technology and automation over the past several decades have made it feasible to perform high-throughput compound screening with cell spheroids, a valuable approach for drug discovery. It is entirely feasible to generate multiple 384-well plates containing adipose spheroids from cryopreserved, single-donor, adipose stem cells, thus incorporating genetic diversity into the discovery stages of research. In this protocol, we describe our method for isolating primary human adipose stem cells and synthesizing cell spheroids comprised of mature adipocytes and stromal cells. Also included are representative outcome measurements useful for characterizing adipocyte metabolism and health. Wherever possible, we describe technologies that can be used to automate characterization and increase throughput.
过去几十年中，技术和自动化的进步使得使用细胞球体进行高通量化合物筛选成为可能，这是一种用于药物发现的有价值的方法。从冷冻保存的、单一供体的脂肪干细胞中生成包含脂肪球体的多个 384 孔板是完全可行的，从而将遗传多样性纳入研究的发现阶段。在本方案中，我们描述了从原代人脂肪干细胞中分离和合成由成熟脂肪细胞和基质细胞组成的细胞球体的方法。还包括用于表征脂肪细胞代谢和健康的代表性结果测量。在可能的情况下，我们描述了可用于自动化表征和提高通量的技术。
